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| − | <div id="seq_features_div" style="clear:both"> <input type="hidden" id="new_dna_format" name="new_dna_format" value=""> <div class="snf_h_pane" style="position: relative;"><div class="snf_h_control_pane" style="position: relative; background-color: rgb(221, 221, 221); padding: 5px; border-bottom: 1px solid gray; font-family: arial, sans-serif; top: 0px; left: 0px; height: 16px;"><span class="format_control" style="position: relative;"><span class="choice disabled" style="position: relative;">Subparts</span><span class="choice" style="position: relative;">|</span><span class="choice selected" style="position: relative;">Ruler</span><span class="choice" style="position: relative;">|</span><span class="choice" style="position: relative;">SS</span><span class="choice" style="position: relative;">|</span><span class="choice" style="position: relative;">DS</span></span><span class="scar_control" style="position: relative;"><span class="legend" style="position: relative;">Scars: </span><span class="choice" style="position: relative;">Show</span><span class="choice" style="position: relative;">|</span><span class="choice selected" style="position: relative;">Hide</span></span><span class="vertical_control disabled" style="position: relative;"><span class="legend" style="position: relative;">Vertical: </span><span class="choice" style="position: relative;">Show</span><span class="choice" style="position: relative;">|</span><span class="choice selected" style="position: relative;">Hide</span></span><span class="SnF_partSeqLength legend" style="position: relative;">Length: 303 bp</span><span class="link" style="position: relative;">Get part sequence</span><span class="link" style="position: relative;">View plasmid<img src="https://parts.igem.org/images/labgenius_p_icon.png" style="width: 15px; height: 15px; position: relative; top: -2px; padding-left: 5px;"></span></div><div class="snf_ss_ds_div" style="position: relative; display: none;"><span style="position: relative; white-space: pre; color: black; font: 12px / 12px monospace;">1 11 21 31 41 51 61 71 81 91</span><div class="rowDiv" style="position: relative; top: 0px; left: 0px; z-index: 40;"><div class="bpDiv" style="position: relative;"><div class="indexDiv" style="position: absolute; font: 12px / 12px monospace; top: 2px; left: -35px; z-index: 60;">1</div><div style="position: relative; font: 12px / 12px monospace; z-index: 60;">tccgataccg gtcgtccgtt tgtggaaatg tacagcgaaa tcccagaaat catccacatg accgaaggtc gtgaactggt tatcccgtgc cgtgtgacca</div><div class="snf_ds_div" style="position: relative; font: 12px / 12px monospace; z-index: 60; display: none;">aggctatggc cagcaggcaa acacctttac atgtcgcttt agggtcttta gtaggtgtac tggcttccag cacttgacca atagggcacg gcacactggt</div><div style="position: relative; height: 5px;"></div></div><div class="featureDiv" style="position: relative; top: 1px; width: 100px; z-index: 60; height: 29px;"><img src="https://parts.igem.org/images/f_icons/brick_l.png" style="top: 0px; left: 1px; height: 7px; width: 3px; position: absolute; z-index: 60;"><img src="https://parts.igem.org/images/f_icons/arrow_r.png" style="top: 0px; left: 648.446px; height: 5px; width: 7px; position: absolute; z-index: 60;"><img src="https://parts.igem.org/images/f_icons/brick_m.png" style="top: 0px; left: 4px; height: 7px; width: 642.446px; position: absolute; z-index: 60;"><div style="font-size: 85%; white-space: nowrap; top: 7px; left: 307.117px; height: 11px; position: absolute; z-index: 60;">VEGFR1D2</div></div></div><div class="rowDiv" style="position: relative; top: 0px; left: 0px; z-index: 40;"><div class="bpDiv" style="position: relative;"><div class="indexDiv" style="position: absolute; font: 12px / 12px monospace; top: 2px; left: -35px; z-index: 60;">101</div><div style="position: relative; font: 12px / 12px monospace; z-index: 60;">gcccaaacat caccgtgacc ctgaaaaaat tcccgctgga taccctgatt ccggatggta aacgtatcat ctgggattct cgtaaaggct ttattatttc</div><div class="snf_ds_div" style="position: relative; font: 12px / 12px monospace; z-index: 60; display: none;">cgggtttgta gtggcactgg gactttttta agggcgacct atgggactaa ggcctaccat ttgcatagta gaccctaaga gcatttccga aataataaag</div><div style="position: relative; height: 5px;"></div></div><div class="featureDiv" style="position: relative; top: 1px; width: 100px; z-index: 60; height: 29px;"><img src="https://parts.igem.org/images/f_icons/arrow_r.png" style="top: 0px; left: -8px; height: 5px; width: 7px; position: absolute; z-index: 60;"><img src="https://parts.igem.org/images/f_icons/arrow_r.png" style="top: 0px; left: 648.446px; height: 5px; width: 7px; position: absolute; z-index: 60;"><img src="https://parts.igem.org/images/f_icons/brick_m.png" style="top: 0px; left: 1px; height: 7px; width: 645.446px; position: absolute; z-index: 60;"><div style="font-size: 85%; white-space: nowrap; top: 7px; left: 307.117px; height: 11px; position: absolute; z-index: 60;">VEGFR1D2</div></div></div><div class="rowDiv" style="position: relative; top: 0px; left: 0px; z-index: 40;"><div class="bpDiv" style="position: relative;"><div class="indexDiv" style="position: absolute; font: 12px / 12px monospace; top: 2px; left: -35px; z-index: 60;">201</div><div style="position: relative; font: 12px / 12px monospace; z-index: 60;">caacgcgacc tacaaagaaa tcggtctgct gacctgcgaa gcaacagtta acggccacct gtacaaaaca aactatctga cccaccgcca gaccaacacc</div><div class="snf_ds_div" style="position: relative; font: 12px / 12px monospace; z-index: 60; display: none;">gttgcgctgg atgtttcttt agccagacga ctggacgctt cgttgtcaat tgccggtgga catgttttgt ttgatagact gggtggcggt ctggttgtgg</div><div style="position: relative; height: 5px;"></div></div><div class="featureDiv" style="position: relative; top: 1px; width: 100px; z-index: 60; height: 29px;"><img src="https://parts.igem.org/images/f_icons/arrow_r.png" style="top: 0px; left: -8px; height: 5px; width: 7px; position: absolute; z-index: 60;"><img src="https://parts.igem.org/images/f_icons/arrow_r.png" style="top: 0px; left: 648.446px; height: 5px; width: 7px; position: absolute; z-index: 60;"><img src="https://parts.igem.org/images/f_icons/brick_m.png" style="top: 0px; left: 1px; height: 7px; width: 645.446px; position: absolute; z-index: 60;"><div style="font-size: 85%; white-space: nowrap; top: 7px; left: 307.117px; height: 11px; position: absolute; z-index: 60;">VEGFR1D2</div></div></div><div class="rowDiv" style="position: relative; top: 0px; left: 0px; z-index: 40;"><div class="bpDiv" style="position: relative;"><div class="indexDiv" style="position: absolute; font: 12px / 12px monospace; top: 2px; left: -35px; z-index: 60;">301</div><div style="position: relative; font: 12px / 12px monospace; z-index: 60;">atc</div><div class="snf_ds_div" style="position: relative; font: 12px / 12px monospace; z-index: 60; display: none;">tag</div><div style="position: relative; height: 5px;"></div></div><div class="featureDiv" style="position: relative; top: 1px; width: 100px; z-index: 60; height: 29px;"><img src="https://parts.igem.org/images/f_icons/arrow_r.png" style="top: 0px; left: -8px; height: 5px; width: 7px; position: absolute; z-index: 60;"><img src="https://parts.igem.org/images/f_icons/brick_r.png" style="top: 0px; left: 14.7921px; height: 7px; width: 3px; position: absolute; z-index: 60;"><img src="https://parts.igem.org/images/f_icons/brick_m.png" style="top: 0px; left: 1px; height: 7px; width: 13.7921px; position: absolute; z-index: 60;"><div style="font-size: 85%; white-space: nowrap; top: 7px; left: 0px; height: 11px; position: absolute; z-index: 60;">VEGFR1D2</div></div></div></div><div class="snf_fw_div" style="position: relative; display: block;"><div style="position: relative; display: block; white-space: pre; color: black; font: 12px / 12px monospace;">1</div><div style="position: relative; left: 0px; height: 18px; width: 647.446px;"><div style="position: absolute; border-top: 1px solid black; top: 4px; left: 0px; height: 1px; width: 647.446px;"><div style="position: absolute; background-color: black; top: 0px; left: 0px; height: 9px; width: 1px;"></div><div style="position: absolute; background-color: black; top: 0px; left: 32px; height: 5px; width: 1px;"></div><div style="position: absolute; background-color: black; top: 0px; left: 65px; height: 5px; width: 1px;"></div><div style="position: absolute; background-color: black; top: 0px; left: 97px; height: 5px; width: 1px;"></div><div style="position: absolute; background-color: black; top: 0px; left: 129px; height: 5px; width: 1px;"></div><div style="position: absolute; background-color: black; top: 0px; left: 162px; height: 9px; width: 1px;"></div><div style="position: absolute; background-color: black; top: 0px; left: 194px; height: 5px; width: 1px;"></div><div style="position: absolute; background-color: black; top: 0px; left: 226px; height: 5px; width: 1px;"></div><div style="position: absolute; background-color: black; top: 0px; left: 259px; height: 5px; width: 1px;"></div><div style="position: absolute; background-color: black; top: 0px; left: 291px; height: 5px; width: 1px;"></div><div style="position: absolute; background-color: black; top: 0px; left: 323px; height: 9px; width: 1px;"></div><div style="position: absolute; background-color: black; top: 0px; left: 356px; height: 5px; width: 1px;"></div><div style="position: absolute; background-color: black; top: 0px; left: 388px; height: 5px; width: 1px;"></div><div style="position: absolute; background-color: black; top: 0px; left: 420px; height: 5px; width: 1px;"></div><div style="position: absolute; background-color: black; top: 0px; left: 453px; height: 5px; width: 1px;"></div><div style="position: absolute; background-color: black; top: 0px; left: 485px; height: 9px; width: 1px;"></div><div style="position: absolute; background-color: black; top: 0px; left: 517px; height: 5px; width: 1px;"></div><div style="position: absolute; background-color: black; top: 0px; left: 549px; height: 5px; width: 1px;"></div><div style="position: absolute; background-color: black; top: 0px; left: 582px; height: 5px; width: 1px;"></div><div style="position: absolute; background-color: black; top: 0px; left: 614px; height: 5px; width: 1px;"></div><div style="position: absolute; background-color: black; top: 0px; left: 646px; height: 9px; width: 1px;"></div></div></div><div class="fw_div" style="position: relative; top: 0px; left: 0px; z-index: 40;"><div class="featureDiv" style="position: relative; top: 1px; width: 100px; z-index: 60; height: 29px;"><img src="https://parts.igem.org/images/f_icons/brick_l.png" style="top: 0px; left: 1px; height: 7px; width: 3px; position: absolute; z-index: 60;"><img src="https://parts.igem.org/images/f_icons/brick_r.png" style="top: 0px; left: 486px; height: 7px; width: 3px; position: absolute; z-index: 60;"><img src="https://parts.igem.org/images/f_icons/brick_m.png" style="top: 0px; left: 4px; height: 7px; width: 482px; position: absolute; z-index: 60;"><div style="font-size: 85%; white-space: nowrap; top: 7px; left: 228.394px; height: 11px; position: absolute; z-index: 60;">VEGFR1D2</div><img src="https://parts.igem.org/images/f_icons/misc_p_m.png" style="top: 0px; left: 0px; height: 5px; width: 7px; position: absolute; z-index: 60;"></div></div></div></div><div id="assembly_compatibility" style="position:relative;clear:both;width:100%">
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| − | <p><style type="text/css">
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| − | .compatibility_div ul,
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| − | .compatibility_div li {
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| − | display: inline;
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| − | }
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| − | .compatibility_div li {
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| − | position: relative;
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| − | padding-top: 2px;
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| − | margin-right:2px;
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| − | }
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| − | .compatibility_div .box {
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| − | top: 35px;
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| − | width: 200px;
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| − | left: 0px;
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| − | }
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| − | .compatibility_div .box_white {
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| − | border: 1px solid gray;
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| − | background-color: white;
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| − | }
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| − | .compatibility_div .box_red {
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| − | border: 1px solid #dd6666;
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| − | background-color: #ffcccc;
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| − | background-image: url('https://parts.igem.org/images/red_not/red_box.png');
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| − | background-repeat: none;
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| − | }
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| − | .compatibility_div .box_green {
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| − | border: 1px solid #44ee44;
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| − | background-color: #aaffaa;
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| − | }
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| − | </p><p><br />
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| − | </p>
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| − | </style></p><div class="compatibility_div" style="display:inline">
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| − | Assembly Compatibility:<ul>
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| − | <li class="boxctrl box_green">10<div class="box"><div>COMPATIBLE WITH RFC[10]</div></div></li>
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| − | <li class="boxctrl box_green">12<div class="box"><div>COMPATIBLE WITH RFC[12]</div></div></li>
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| − | <li class="boxctrl box_green">21<div class="box"><div>COMPATIBLE WITH RFC[21]</div></div></li>
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| − | <li class="boxctrl box_green">23<div class="box"><div>COMPATIBLE WITH RFC[23]</div></div></li>
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| − | <li class="boxctrl box_red">25<div class="box"><div style="border-bottom:1px solid gray;">INCOMPATIBLE WITH RFC[25]</div>Illegal AgeI site found at 7<br></div></li>
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| − | <li class="boxctrl box_green">1000<div class="box"><div>COMPATIBLE WITH RFC[1000]</div></div></li>
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| − | </ul>
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| − | </div></div> <div class="snf_c_pane" style="position: relative; display: none;"><div id="c_edit_pane" style="width:940px;margin-top:5px;min-height:50px;"><div id="subpart_pane" style="padding: 2px 4px 2px 2px; width: 600px; border: none;">
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| − | <theader></theader><table id="subpart_table" style="border: 1px solid gray" class="treetable"><tbody></tbody></table></div></div></div></div>
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| | VEGFR1D2: | | VEGFR1D2: |
| | VEGF binds to two tyrosine kinase receptors, VEGF receptor 1 (VEGFR1/Flt-1) and receptor 2 (VEGFR2/KDR), which are localized on the cell surface of various cell types. The receptors are organized in two intracellular kinase domains, a short transmembrane region and an extracellular portion constituted of seven immunoglobulin-like domains. The binding of VEGF to the extracellular domain induces receptor dimerization, transphosphorylation of the kinase domains, and activation of the intracellular signaling. Deletion studies have shown that the ligand binding function resides within the first three domains of VEGFR14 and in domains 2 and 3 of VEGFR2. Deletion of the second extracellular domain of VEGFR1 (VEGFR1D2) completely abolishes the binding to VEGF, whereas VEGFR1D2 binds VEGF with an affinity that is only 60 times lower than the entire extracellular portion of the receptor. | | VEGF binds to two tyrosine kinase receptors, VEGF receptor 1 (VEGFR1/Flt-1) and receptor 2 (VEGFR2/KDR), which are localized on the cell surface of various cell types. The receptors are organized in two intracellular kinase domains, a short transmembrane region and an extracellular portion constituted of seven immunoglobulin-like domains. The binding of VEGF to the extracellular domain induces receptor dimerization, transphosphorylation of the kinase domains, and activation of the intracellular signaling. Deletion studies have shown that the ligand binding function resides within the first three domains of VEGFR14 and in domains 2 and 3 of VEGFR2. Deletion of the second extracellular domain of VEGFR1 (VEGFR1D2) completely abolishes the binding to VEGF, whereas VEGFR1D2 binds VEGF with an affinity that is only 60 times lower than the entire extracellular portion of the receptor. |
VEGFR1D2:
VEGF binds to two tyrosine kinase receptors, VEGF receptor 1 (VEGFR1/Flt-1) and receptor 2 (VEGFR2/KDR), which are localized on the cell surface of various cell types. The receptors are organized in two intracellular kinase domains, a short transmembrane region and an extracellular portion constituted of seven immunoglobulin-like domains. The binding of VEGF to the extracellular domain induces receptor dimerization, transphosphorylation of the kinase domains, and activation of the intracellular signaling. Deletion studies have shown that the ligand binding function resides within the first three domains of VEGFR14 and in domains 2 and 3 of VEGFR2. Deletion of the second extracellular domain of VEGFR1 (VEGFR1D2) completely abolishes the binding to VEGF, whereas VEGFR1D2 binds VEGF with an affinity that is only 60 times lower than the entire extracellular portion of the receptor.
The equilibrium constant for the binding of VEGF to one VEGFR1D2 molecule was determined by surface plasmon resonance. The obtained value, 2.9 ± 0.2 nM, is consistent with the constant reported for the binding of VEGF to monomeric VEGFR1D2.
We used pBBR plasmid as a backbone and transfered VEGFR1D2 into Escherichia coli Nissle 1917, and finally succeeded in VEGFR1D2-expressing.
