Plasmid_Backbone

Part:BBa_K3728002

Designed by: Eric Shih   Group: iGEM21_Mingdao   (2021-08-19)
Revision as of 04:12, 25 August 2021 by Eric2005 (Talk | contribs)


/* Reset italic styling set by user agent */ cite, dfn { font-style: inherit; } /* Straight quote marks for */ q { quotes: '"' '"' "'" "'"; } /* Avoid collision of blockquote with floating elements by swapping margin and padding */ blockquote { overflow: hidden; margin: 1em 0; padding: 0 40px; } /* Consistent size for , and */ small { font-size: 85%; } .mw-body-content sub, .mw-body-content sup, span.reference /* for Parsoid */ { font-size: 80%; } /* Same spacing for indented and unindented paragraphs on talk pages */ .ns-talk .mw-body-content dd { margin-top: 0.4em; margin-bottom: 0.4em; } /* Main page fixes */ #interwiki-completelist { font-weight: bold; } /* Reduce page jumps by hiding collapsed/dismissed content */ .client-js .mw-special-Watchlist #watchlist-message, .client-js .collapsible:not( .mw-made-collapsible).collapsed > tbody > tr:not(:first-child), /* Hide charinsert base for those not using the gadget */ #editpage-specialchars { display: none; } /* Adds padding above Watchlist announcements where new recentchanges/watchlist filters are enabled */ .mw-rcfilters-enabled .mw-specialpage-summary { margin-top: 1em; } /* Highlight linked elements (such as clicked references) in blue */ body.action-info .mw-body-content :target, .citation:target { background-color: rgba(0, 127, 255, 0.133); } /* Styling for citations. Breaks long urls, etc., rather than overflowing box */ .citation { word-wrap: break-word; } /* Make the list of references smaller * Keep in sync with Template:Refbegin/styles.css * And Template:Reflist/styles.css */ ol.references { font-size: 90%; margin-bottom: 0.5em; } /* Style for horizontal lists (separator following item). @source mediawiki.org/wiki/Snippets/Horizontal_lists @revision 8 (2016-05-21) @author [[User:Edokter]] */ .hlist dl, .hlist ol, .hlist ul { margin: 0; padding: 0; } /* Display list items inline */ .hlist dd, .hlist dt, .hlist li { margin: 0; /* don't trust the note that says margin doesn't work with inline * removing margin: 0 makes dds have margins again */ display: inline; } /* Display nested lists inline */ .hlist.inline, .hlist.inline dl, .hlist.inline ol, .hlist.inline ul, .hlist dl dl, .hlist dl ol, .hlist dl ul, .hlist ol dl, .hlist ol ol, .hlist ol ul, .hlist ul dl, .hlist ul ol, .hlist ul ul { display: inline; } /* Hide empty list items */ .hlist .mw-empty-li { display: none; } /* Generate interpuncts */ .hlist dt:after { content: ": "; } /** * Note hlist style usage differs in Minerva and is defined in core as well! * Please check Minerva desktop (and Minerva.css) when changing * See https://phabricator.wikimedia.org/T213239 */ .hlist dd:after, .hlist li:after { content: " · "; font-weight: bold; } .hlist dd:last-child:after, .hlist dt:last-child:after, .hlist li:last-child:after { content: none; } /* Add parentheses around nested lists */ .hlist dd dd:first-child:before, .hlist dd dt:first-child:before, .hlist dd li:first-child:before, .hlist dt dd:first-child:before, .hlist dt dt:first-child:before, .hlist dt li:first-child:before, .hlist li dd:first-child:before, .hlist li dt:first-child:before, .hlist li li:first-child:before { content: " ("; font-weight: normal; } .hlist dd dd:last-child:after, .hlist dd dt:last-child:after, .hlist dd li:last-child:after, .hlist dt dd:last-child:after, .hlist dt dt:last-child:after, .hlist dt li:last-child:after, .hlist li dd:last-child:after, .hlist li dt:last-child:after, .hlist li li:last-child:after { content: ")"; font-weight: normal; } /* Put ordinals in front of ordered list items */ .hlist ol { counter-reset: listitem; } .hlist ol > li { counter-increment: listitem; } .hlist ol > li:before { content: " " counter(listitem) "\a0"; } .hlist dd ol > li:first-child:before, .hlist dt ol > li:first-child:before, .hlist li ol > li:first-child:before { content: " (" counter(listitem) "\a0"; } /* Unbulleted lists */ .plainlist ol, .plainlist ul { line-height: inherit; list-style: none none; margin: 0; } .plainlist ol li, .plainlist ul li { margin-bottom: 0; } /* Default style for navigation boxes */ .navbox { /* Navbox container style */ box-sizing: border-box; border: 1px solid #a2a9b1; width: 100%; clear: both; font-size: 88%; text-align: center; padding: 1px; margin: 1em auto 0; /* Prevent preceding content from clinging to navboxes */ } .navbox .navbox { margin-top: 0; /* No top margin for nested navboxes */ } .navbox + .navbox { margin-top: -1px; /* Single pixel border between adjacent navboxes */ } .navbox-inner, .navbox-subgroup { width: 100%; } .navbox-group, .navbox-title, .navbox-abovebelow { padding: 0.25em 1em; /* Title, group and above/below styles */ line-height: 1.5em; text-align: center; } th.navbox-group { /* Group style */ white-space: nowrap; /* @noflip */ text-align: right; } .navbox, .navbox-subgroup { background-color: #fdfdfd; /* Background color */ } .navbox-list { line-height: 1.5em; border-color: #fdfdfd; /* Must match background color */ } /* cell spacing for navbox cells */ tr + tr > .navbox-abovebelow, tr + tr > .navbox-group, tr + tr > .navbox-image, tr + tr > .navbox-list { /* Borders above 2nd, 3rd, etc. rows */ border-top: 2px solid #fdfdfd; /* Must match background color */ } .navbox th, .navbox-title { background-color: #ccccff; /* Level 1 color */ } .navbox-abovebelow, th.navbox-group, .navbox-subgroup .navbox-title { background-color: #ddddff; /* Level 2 color */ } .navbox-subgroup .navbox-group, .navbox-subgroup .navbox-abovebelow { background-color: #e6e6ff; /* Level 3 color */ } .navbox-even { background-color: #f7f7f7; /* Even row striping */ } .navbox-odd { background-color: transparent; /* Odd row striping */ } .navbox .hlist td dl, .navbox .hlist td ol, .navbox .hlist td ul, .navbox td.hlist dl, .navbox td.hlist ol, .navbox td.hlist ul { padding: 0.125em 0; /* Adjust hlist padding in navboxes */ } /* Styling for JQuery makeCollapsible, matching that of collapseButton */ .mw-parser-output .mw-collapsible-toggle { font-weight: normal; /* @noflip */ text-align: right; padding-right: 0.2em; padding-left: 0.2em; } .mw-collapsible-leftside-toggle .mw-collapsible-toggle { /* @noflip */ float: left; /* @noflip */ text-align: left; } /* Infobox template style */ .infobox { border: 1px solid #a2a9b1; border-spacing: 3px; background-color: #f8f9fa; color: black; /* @noflip */ margin: 0.5em 0 0.5em 1em; padding: 0.2em; /* @noflip */ float: right; /* @noflip */ clear: right; font-size: 88%; line-height: 1.5em; width: 22em; } /* not strictly certain these styles are necessary * just replicating the module faithfully */ .infobox-subbox { padding: 0; border: none; margin: -3px; width: auto; min-width: 100%; font-size: 100%; clear: none; float: none; background-color: transparent; } .infobox-header, .infobox-label, .infobox-above, .infobox-full-data, .infobox-data, .infobox-below, .infobox-subheader, .infobox-image, .infobox-navbar, /* Remove element declaration when both a) infobox3cols is migrated and * b) when every .infobox thing is using the standard module/templates */ .infobox th, .infobox td { vertical-align: top; } .infobox-label, .infobox-data, /* Remove element declaration when both a) infobox3cols is migrated and * b) when every .infobox thing is using the standard module/templates */ .infobox th, .infobox td { /* @noflip */ text-align: left; } /* remove .infobox when th, td above are removed */ .infobox .infobox-above, .infobox .infobox-title, /* Remove element declaration when both a) infobox3cols is migrated and * b) when every .infobox thing is using the standard module/templates */ .infobox caption { font-size: 125%; font-weight: bold; text-align: center; } .infobox-title, /* Remove element declaration when both a) infobox3cols is migrated and * b) when every .infobox thing is using the standard module/templates */ .infobox caption { padding: 0.2em; } /* remove .infobox when th, td above are removed */ .infobox .infobox-header, .infobox .infobox-subheader, .infobox .infobox-image, .infobox .infobox-full-data, .infobox .infobox-below { text-align: center; } /* remove .infobox when th, td above are removed */ .infobox .infobox-navbar { /* @noflip */ text-align: right; } .infobox.bordered { border-collapse: collapse; } /* TODO split definitions to appropriate class names when live from HTML element */ .infobox.bordered td, .infobox.bordered th { border: 1px solid #a2a9b1; } /* TODO split definitions to appropriate class names when live from HTML element */ .infobox.bordered .borderless td, .infobox.bordered .borderless th { border: 0; } /* styles for bordered infobox with merged rows */ /* TODO split definitions to appropriate class names when live from HTML element */ .infobox.bordered .mergedtoprow td, .infobox.bordered .mergedtoprow th { border: 0; border-top: 1px solid #a2a9b1; /* @noflip */ border-right: 1px solid #a2a9b1; } /* TODO split definitions to appropriate class names when live from HTML element */ .infobox.bordered .mergedrow td, .infobox.bordered .mergedrow th { border: 0; /* @noflip */ border-right: 1px solid #a2a9b1; } /* Styles for geography infoboxes, eg countries, country subdivisions, cities, etc. */ .infobox.geography { border-collapse: collapse; line-height: 1.2em; font-size: 90%; } /* TODO split definitions to appropriate class names when live from HTML element */ .infobox.geography td, .infobox.geography th { border-top: 1px solid #a2a9b1; padding: 0.4em 0.6em 0.4em 0.6em; } /* TODO split definitions to appropriate class names when live from HTML element */ .infobox.geography .mergedtoprow td, .infobox.geography .mergedtoprow th { border-top: 1px solid #a2a9b1; padding: 0.4em 0.6em 0.2em 0.6em; } /* TODO split definitions to appropriate class names when live from HTML element */ .infobox.geography .mergedrow td, .infobox.geography .mergedrow th { border: 0; padding: 0 0.6em 0.2em 0.6em; } /* TODO split definitions to appropriate class names when live from HTML element */ .infobox.geography .mergedbottomrow td, .infobox.geography .mergedbottomrow th { border-top: 0; border-bottom: 1px solid #a2a9b1; padding: 0 0.6em 0.4em 0.6em; } /* TODO split definitions to appropriate class names when live from HTML element */ .infobox.geography .maptable td, .infobox.geography .maptable th { border: 0; padding: 0; } /* Normal font styling for wikitable row headers with scope="row" tag */ .wikitable.plainrowheaders th[scope=row] { font-weight: normal; /* @noflip */ text-align: left; } /* Lists in wikitable data cells are always left-aligned */ .wikitable td ul, .wikitable td ol, .wikitable td dl { /* @noflip */ text-align: left; } /* ...unless they also use the hlist class */ .toc.hlist ul, #toc.hlist ul, .wikitable.hlist td ul, .wikitable.hlist td ol, .wikitable.hlist td dl { text-align: inherit; } /* Fix for hieroglyphs specificity issue in infoboxes ([[phab:T43869]]) */ table.mw-hiero-table td { vertical-align: middle; } /* Change the external link icon to an Adobe icon for all PDF files */ .mw-parser-output a[href$=".pdf"].external, .mw-parser-output a[href*=".pdf?"].external, .mw-parser-output a[href*=".pdf#"].external, .mw-parser-output a[href$=".PDF"].external, .mw-parser-output a[href*=".PDF?"].external, .mw-parser-output a[href*=".PDF#"].external { background: url("//upload.wikimedia.org/wikipedia/commons/2/23/Icons-mini-file_acrobat.gif") no-repeat right; /* @noflip */ padding-right: 18px; } /* Messagebox templates */ .messagebox { border: 1px solid #a2a9b1; background-color: #f8f9fa; width: 80%; margin: 0 auto 1em auto; padding: .2em; } .messagebox.merge { border: 1px solid #c0b8cc; background-color: #f0e5ff; text-align: center; } .messagebox.cleanup { border: 1px solid #9f9fff; background-color: #efefff; text-align: center; } .messagebox.standard-talk { border: 1px solid #c0c090; background-color: #f8eaba; margin: 4px auto; } /* For old WikiProject banners inside banner shells. */ .mbox-inside .standard-talk { border: 1px solid #c0c090; background-color: #f8eaba; width: 100%; margin: 2px 0; padding: 2px; } .messagebox.small { width: 238px; font-size: 85%; /* @noflip */ float: right; clear: both; /* @noflip */ margin: 0 0 1em 1em; line-height: 1.25em; } .messagebox.small-talk { width: 238px; font-size: 85%; /* @noflip */ float: right; clear: both; /* @noflip */ margin: 0 0 1em 1em; line-height: 1.25em; background-color: #f8eaba; } /* Cell sizes for ambox/tmbox/imbox/cmbox/ombox/fmbox/dmbox message boxes */ th.mbox-text, td.mbox-text { /* The message body cell(s) */ border: none; /* @noflip */ padding: 0.25em 0.9em; /* 0.9em left/right */ width: 100%; /* Make all mboxes the same width regardless of text length */ } td.mbox-image { /* The left image cell */ border: none; /* @noflip */ padding: 2px 0 2px 0.9em; /* 0.9em left, 0px right */ text-align: center; } td.mbox-imageright { /* The right image cell */ border: none; /* @noflip */ padding: 2px 0.9em 2px 0; /* 0px left, 0.9em right */ text-align: center; } td.mbox-empty-cell { /* An empty narrow cell */ border: none; padding: 0; width: 1px; } /* Article message box styles */ table.ambox { margin: 0 10%; /* 10% = Will not overlap with other elements */ border: 1px solid #a2a9b1; /* @noflip */ border-left: 10px solid #36c; /* Default "notice" blue */ background-color: #fbfbfb; box-sizing: border-box; } table.ambox + table.ambox { /* Single border between stacked boxes. */ margin-top: -1px; } .ambox th.mbox-text, .ambox td.mbox-text { /* The message body cell(s) */ padding: 0.25em 0.5em; /* 0.5em left/right */ } .ambox td.mbox-image { /* The left image cell */ /* @noflip */ padding: 2px 0 2px 0.5em; /* 0.5em left, 0px right */ } .ambox td.mbox-imageright { /* The right image cell */ /* @noflip */ padding: 2px 0.5em 2px 0; /* 0px left, 0.5em right */ } table.ambox-notice { /* @noflip */ border-left: 10px solid #36c; /* Blue */ } table.ambox-speedy { /* @noflip */ border-left: 10px solid #b32424; /* Red */ background-color: #fee7e6; /* Pink */ } table.ambox-delete { /* @noflip */ border-left: 10px solid #b32424; /* Red */ } table.ambox-content { /* @noflip */ border-left: 10px solid #f28500; /* Orange */ } table.ambox-style { /* @noflip */ border-left: 10px solid #fc3; /* Yellow */ } table.ambox-move { /* @noflip */ border-left: 10px solid #9932cc; /* Purple */ } table.ambox-protection { /* @noflip */ border-left: 10px solid #a2a9b1; /* Gray-gold */ } /* Image message box styles */ table.imbox { margin: 4px 10%; border-collapse: collapse; border: 3px solid #36c; /* Default "notice" blue */ background-color: #fbfbfb; box-sizing: border-box; } .imbox .mbox-text .imbox { /* For imboxes inside imbox-text cells. */ margin: 0 -0.5em; /* 0.9 - 0.5 = 0.4em left/right. */ display: block; /* Fix for webkit to force 100% width. */ } .mbox-inside .imbox { /* For imboxes inside other templates. */ margin: 4px; } table.imbox-notice { border: 3px solid #36c; /* Blue */ } table.imbox-speedy { border: 3px solid #b32424; /* Red */ background-color: #fee7e6; /* Pink */ } table.imbox-delete { border: 3px solid #b32424; /* Red */ } table.imbox-content { border: 3px solid #f28500; /* Orange */ } table.imbox-style { border: 3px solid #fc3; /* Yellow */ } table.imbox-move { border: 3px solid #9932cc; /* Purple */ } table.imbox-protection { border: 3px solid #a2a9b1; /* Gray-gold */ } table.imbox-license { border: 3px solid #88a; /* Dark gray */ background-color: #f7f8ff; /* Light gray */ } table.imbox-featured { border: 3px solid #cba135; /* Brown-gold */ } /* Category message box styles */ table.cmbox { margin: 3px 10%; border-collapse: collapse; border: 1px solid #a2a9b1; background-color: #dfe8ff; /* Default "notice" blue */ box-sizing: border-box; } table.cmbox-notice { background-color: #d8e8ff; /* Blue */ } table.cmbox-speedy { margin-top: 4px; margin-bottom: 4px; border: 4px solid #b32424; /* Red */ background-color: #ffdbdb; /* Pink */ } table.cmbox-delete { background-color: #ffdbdb; /* Pink */ } table.cmbox-content { background-color: #ffe7ce; /* Orange */ } table.cmbox-style { background-color: #fff9db; /* Yellow */ } table.cmbox-move { background-color: #e4d8ff; /* Purple */ } table.cmbox-protection { background-color: #efefe1; /* Gray-gold */ } /* Other pages message box styles */ table.ombox { margin: 4px 10%; border-collapse: collapse; border: 1px solid #a2a9b1; /* Default "notice" gray */ background-color: #f8f9fa; box-sizing: border-box; } table.ombox-notice { border: 1px solid #a2a9b1; /* Gray */ } table.ombox-speedy { border: 2px solid #b32424; /* Red */ background-color: #fee7e6; /* Pink */ } table.ombox-delete { border: 2px solid #b32424; /* Red */ } table.ombox-content { border: 1px solid #f28500; /* Orange */ } table.ombox-style { border: 1px solid #fc3; /* Yellow */ } table.ombox-move { border: 1px solid #9932cc; /* Purple */ } table.ombox-protection { border: 2px solid #a2a9b1; /* Gray-gold */ } /* Talk page message box styles */ table.tmbox { margin: 4px 10%; border-collapse: collapse; border: 1px solid #c0c090; /* Default "notice" gray-brown */ background-color: #f8eaba; min-width: 80%; box-sizing: border-box; } .tmbox.mbox-small { min-width: 0; /* reset the min-width of tmbox above */ } .mediawiki .mbox-inside .tmbox { /* For tmboxes inside other templates. The "mediawiki" class ensures that */ margin: 2px 0; /* this declaration overrides other styles (including mbox-small above) */ width: 100%; /* For Safari and Opera */ } .mbox-inside .tmbox.mbox-small { /* "small" tmboxes should not be small when */ line-height: 1.5em; /* also "nested", so reset styles that are */ font-size: 100%; /* set in "mbox-small" above. */ } table.tmbox-speedy { border: 2px solid #b32424; /* Red */ background-color: #fee7e6; /* Pink */ } table.tmbox-delete { border: 2px solid #b32424; /* Red */ } table.tmbox-content { border: 2px solid #f28500; /* Orange */ } table.tmbox-style { border: 2px solid #fc3; /* Yellow */ } table.tmbox-move { border: 2px solid #9932cc; /* Purple */ } table.tmbox-protection, table.tmbox-notice { border: 1px solid #c0c090; /* Gray-brown */ } /* Footer and header message box styles */ table.fmbox { clear: both; margin: 0.2em 0; width: 100%; border: 1px solid #a2a9b1; background-color: #f8f9fa; /* Default "system" gray */ box-sizing: border-box; } table.fmbox-system { background-color: #f8f9fa; } table.fmbox-warning { border: 1px solid #bb7070; /* Dark pink */ background-color: #ffdbdb; /* Pink */ } table.fmbox-editnotice { background-color: transparent; } /* Div based "warning" style fmbox messages. */ div.mw-warning-with-logexcerpt, div.mw-lag-warn-high, div.mw-cascadeprotectedwarning, div#mw-protect-cascadeon, div.titleblacklist-warning, div.locked-warning { clear: both; margin: 0.2em 0; border: 1px solid #bb7070; background-color: #ffdbdb; padding: 0.25em 0.9em; box-sizing: border-box; } /* Use default color for partial block fmbox banner per [[Special:PermaLink/1028105567#pblock-style]] */ .mw-contributions-blocked-notice-partial .mw-warning-with-logexcerpt { border-color: #fc3; background-color: #fef6e7; } /* These mbox-small classes must be placed after all other ambox/tmbox/ombox etc classes. "html body.mediawiki" is so they override "table.ambox + table.ambox" above. */ html body.mediawiki .mbox-small { /* For the "small=yes" option. */ /* @noflip */ clear: right; /* @noflip */ float: right; /* @noflip */ margin: 4px 0 4px 1em; box-sizing: border-box; width: 238px; font-size: 88%; line-height: 1.25em; } html body.mediawiki .mbox-small-left { /* For the "small=left" option. */ /* @noflip */ margin: 4px 1em 4px 0; box-sizing: border-box; overflow: hidden; width: 238px; border-collapse: collapse; font-size: 88%; line-height: 1.25em; } /* Style for compact ambox */ /* Hide the images */ .compact-ambox table .mbox-image, .compact-ambox table .mbox-imageright, .compact-ambox table .mbox-empty-cell { display: none; } /* Remove borders, backgrounds, padding, etc. */ .compact-ambox table.ambox { border: none; border-collapse: collapse; background-color: transparent; margin: 0 0 0 1.6em !important; padding: 0 !important; width: auto; display: block; } body.mediawiki .compact-ambox table.mbox-small-left { font-size: 100%; width: auto; margin: 0; } /* Style the text cell as a list item and remove its padding */ .compact-ambox table .mbox-text { padding: 0 !important; margin: 0 !important; } .compact-ambox table .mbox-text-span { display: list-item; line-height: 1.5em; list-style-type: square; list-style-image: url(/w/skins/MonoBook/resources/images/bullet.svg); } /* Allow for hiding text in compact form */ .compact-ambox .hide-when-compact { display: none; } /* Remove underlines from certain links */ .nounderlines a, .IPA a:link, .IPA a:visited { text-decoration: none !important; } /* Prevent line breaks in silly places where desired (nowrap) and links when we don't want them to (nowraplinks a) */ .nowrap, .nowraplinks a { white-space: nowrap; } /* But allow wrapping where desired: */ .wrap, .wraplinks a { white-space: normal; } /* Increase the height of the image upload box */ #wpUploadDescription { height: 13em; } /* Minimum thumb width */ .thumbinner { min-width: 100px; } /* Prevent floating boxes from overlapping any category listings, file histories, edit previews, and edit [Show changes] views. */ #mw-subcategories, #mw-pages, #mw-category-media, #filehistory, #wikiPreview, #wikiDiff { clear: both; } /* Selectively hide headers in WikiProject banners */ /* TemplateStyles */ .wpb .wpb-header { display: none; } .wpbs-inner .wpb .wpb-header { display: table-row; } .wpbs-inner .wpb-outside { display: none; /* hide things that should only display outside shells */ } /* Styling for Abuse Filter tags */ .mw-tag-markers { font-style: italic; font-size: 90%; } /* Hide stuff meant for accounts with special permissions. Made visible again in [[MediaWiki:Group-checkuser.css]], [[MediaWiki:Group-sysop.css]], [[MediaWiki:Group-patroller.css]], [[MediaWiki:Group-templateeditor.css]], [[MediaWiki:Group-extendedmover.css]], [[MediaWiki:Group-extendedconfirmed.css]], and [[Mediawiki:Group-autoconfirmed.css]]. */ .checkuser-show, .sysop-show, .patroller-show, .templateeditor-show, .extendedmover-show, .extendedconfirmed-show, .autoconfirmed-show, .user-show { display: none; } /* Hide the redlink generated by {{Editnotice}}, this overrides the ".sysop-show { display: none; }" above that applies to the same link as well. See [[phab:T45013]] Hide the images in editnotices to keep them readable in VE view. Long term, editnotices should become a core feature so that they can be designed responsive. */ .ve-ui-mwNoticesPopupTool-item .editnotice-redlink, .ve-ui-mwNoticesPopupTool-item .mbox-image, .ve-ui-mwNoticesPopupTool-item .mbox-imageright { display: none !important; } /* Remove bullets when there are multiple edit page warnings */ ul.permissions-errors > li { list-style: none none; } ul.permissions-errors { margin: 0; } /* texhtml class for inline math (based on generic times-serif class) */ span.texhtml { font-family: "Nimbus Roman No9 L", "Times New Roman", Times, serif; font-size: 118%; line-height: 1; white-space: nowrap; /* Force tabular and lining display for texhtml */ -moz-font-feature-settings: "lnum", "tnum", "kern" 0; -webkit-font-feature-settings: "lnum", "tnum", "kern" 0; font-feature-settings: "lnum", "tnum", "kern" 0; font-variant-numeric: lining-nums tabular-nums; font-kerning: none; } span.texhtml span.texhtml { font-size: 100%; } span.mwe-math-mathml-inline { font-size: 118%; } /* Make be left aligned with one space indent for * compatibility with style conventions */ .mwe-math-fallback-image-display, .mwe-math-mathml-display { margin-left: 1.6em !important; margin-top: 0.6em; margin-bottom: 0.6em; } .mwe-math-mathml-display math { display: inline; } /* Work-around for [[phab:T25965]] / [[phab:T100106]] (Kaltura advertisement) */ .k-player .k-attribution { visibility: hidden; } /* Move 'play' button of video player to bottom left corner */ .PopUpMediaTransform a .play-btn-large { margin: 0; top: auto; right: auto; bottom: 0; left: 0; } /* Force imgs in galleries to have borders by wrapping them in class=bordered-images */ .bordered-images img { border: solid #ddd 1px; } @media screen { /* Gallery styles background changes are restricted to screen view. In printing we should avoid applying backgrounds. */ /* The backgrounds for galleries. */ #content .gallerybox div.thumb { /* Light gray padding */ background-color: #f8f9fa; } /* Put a chequered background behind images, only visible if they have transparency. '.filehistory a img' and '#file img:hover' are handled by MediaWiki core (as of 1.19) */ .gallerybox .thumb img { background: #fff url(//upload.wikimedia.org/wikipedia/commons/5/5d/Checker-16x16.png) repeat; } /* But not on articles, user pages, portals or with opt-out. */ .ns-0 .gallerybox .thumb img, .ns-2 .gallerybox .thumb img, .ns-100 .gallerybox .thumb img, .nochecker .gallerybox .thumb img { background-image: none; } /* Display "From Wikipedia, the free encyclopedia" in skins that support it, do not apply to print mode */ #siteSub { display: block; } } /* Hide FlaggedRevs notice UI when there are no pending changes */ .flaggedrevs_draft_synced, .flaggedrevs_stable_synced, /* "Temporary" to remove links in sidebar T255381 */ #t-upload, /* Hide broken download box on Special:Book pending T285400 */ .mw-special-Book #coll-downloadbox { display: none; }


pTol2: a Tol2 vector - the Tol2 transposable element



T--Mingdao--BBa K3728002-1.png

ThisTol2 transposon system is highly used in zebrafish transgenesis. The transposase protein (TPase) is from the Medaka fish (Oryzias latipes) aka Japanese rice fish, which catalyzes the transposition of the Tol2 elements through cut-and-paste mechanism. The minimal transposable Tol2 sequence (mTol2) contains 200-bp left arm and 150-bp right arm. Up to 11kb DNA insert between Tol2 sequence can be integrated into the genome of nearly all vertebrates including zebrafish, frog, chicken, mouse, and human [1].

ThisA further application in synthetic biology was demonstrated by Jun Ni, et. al.[2], in which the recombinant TPase protein is fully functional in HeLa cell line and Zebrafish germline cells. In addition, the TPase can be expressed under T7 promoter in E. coli BL21 and purified with N-terminal 6xHis tag. The transposase is active in vitro and mediated the integration of DNA fragments between plasmids with Tol2 elements.

ThisIn our study, we constructed BioBrick Parts of T7-TPase (BBa_K3728001) and the BioBrick compatible Tol2 vectors (BBa_K3728003) with reporter (KanR:BBa_K3728004; GFP:BBa_K3728005; RFP:BBa_K3728006; amilCP:BBa_K3728007) and Phi29 DNA polymerase genes (BBa_K3728008). We prepared the In vitro transcription-translation (TXTL) system [3][4]and expressed the functional reporter proteins. The recombinant TPase and Phil29 DNA polymerase with His tag were expressed in E. coli BL21. The purified proteins were functional in the plasmid integration assay and rolling circle amplification(RCA) application, respectively.


CONSTRUCTION – BIOBRICK COMPATIBLE VECTOR

CHARACTERIZATION – PLASMID INTEGRATION

APPLICATION – PHAGE ENGINEERING

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3452
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 29
    Illegal NotI site found at 3458
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3452
    Illegal BglII site found at 3340
    Illegal XhoI site found at 3419
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 3452
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 3452
    Plasmid lacks a suffix.
    Illegal XbaI site found at 3467
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NgoMIV site found at 642
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI site found at 1818
    Illegal SapI.rc site found at 2900


REFERENCE

  1. Kawakami K. Tol2: a versatile gene transfer vector in vertebrates. Genome Biol. 2007;8 Suppl 1(Suppl 1):S7. doi: 10.1186/gb-2007-8-s1-s7
  2. Ni J, Wangensteen KJ, Nelsen D, Balciunas D, Skuster KJ, Urban MD, Ekker SC. Active recombinant Tol2 transposase for gene transfer and gene discovery applications. Mob DNA. 2016 Mar 31;7:6. doi: 10.1186/s13100-016-0062-z.
  3. Garenne D, Noireaux V. Cell-free transcription-translation: engineering biology from the nanometer to the millimeter scale. Curr Opin Biotechnol. 2019 Aug;58:19-27. doi: 10.1016/j.copbio.2018.10.007.
  4. Rustad M, Eastlund A, Marshall R, Jardine P, Noireaux V. Synthesis of Infectious Bacteriophages in an E. coli-based Cell-free Expression System. J Vis Exp. 2017 Aug 17;(126):56144. doi: 10.3791/56144.
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