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| ===Design Notes=== | | ===Design Notes=== |
− | Freiburg standard25 part
| + | Designed with Biobrick pre-and suffix for fusion proteins according to the [https://parts.igem.org/Assembly_standard_25 RFC 25].<br> |
| + | [https://parts.igem.org/Image:DigA.txt Commented GenBank file] |
| | | |
| ===Source=== | | ===Source=== |
| | | |
− | synthesized by purimex | + | Planned and designed by Team Freiburg Bioware09 and synthesized by purimex.<br> |
− | | + | |
− | LOCUS His+FluA+Split+F 3616 bp DNA circular SYN 08-OCT-2009
| + | |
− | DEFINITION Ligation of His+FluA+Split+FokI into pMA2_NgoMIV_SpeI
| + | |
− | ACCESSION His+FluA+Split+F
| + | |
− | KEYWORDS .
| + | |
− | SOURCE Unknown.
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− | ORGANISM Unknown
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− | Unclassified.
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− | REFERENCE 1 (bases 1 to 3616)
| + | |
− | AUTHORS Self
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− | JOURNAL Unpublished.
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− | COMMENT SECID/File created by SciEd Central, Scientific & Educational Software
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− | COMMENT SECNOTES|Vector molecule: pMA2_NgoMIV_SpeI
| + | |
− | Fragment ends: SpeI and NgoMIV
| + | |
− | Fragment size: 2412
| + | |
− | Insert molecule: His+FluA+Split+FokI
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− | Fragment ends: NgoMIV and SpeI
| + | |
− | Fragment size: 1204
| + | |
− | FEATURES Location/Qualifiers
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− | CDS 129..989
| + | |
− | /gene="AmpR"
| + | |
− | /SECDrawAs="Gene"
| + | |
− | CDS 2008..3213
| + | |
− | /gene="ORF-1"
| + | |
− | /SECDrawAs="Gene"
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− | CDS 2017..2034
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− | /gene="His Tag"
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− | /SECDrawAs="Gene"
| + | |
− | CDS 2041..2562
| + | |
− | /gene="FluA"
| + | |
− | /SECDrawAs="Gene"
| + | |
− | CDS 2569..2619
| + | |
− | /gene="Split-Linker"
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− | /SECDrawAs="Gene"
| + | |
− | CDS 2623..3212
| + | |
− | /gene="'Fok inaktiv'"
| + | |
− | /SECDrawAs="Gene"
| + | |
− | ORIGIN
| + | |
− | 1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
| + | |
− | 61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
| + | |
− | 121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
| + | |
− | 181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
| + | |
− | 241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
| + | |
− | 301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
| + | |
− | 361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
| + | |
− | 421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
| + | |
− | 481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
| + | |
− | 541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
| + | |
− | 601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
| + | |
− | 661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
| + | |
− | 721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
| + | |
− | 781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
| + | |
− | 841 gtggttctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
| + | |
− | 901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
| + | |
− | 961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
| + | |
− | 1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
| + | |
− | 1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
| + | |
− | 1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
| + | |
− | 1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
| + | |
− | 1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
| + | |
− | 1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
| + | |
− | 1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
| + | |
− | 1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
| + | |
− | 1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
| + | |
− | 1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
| + | |
− | 1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
| + | |
− | 1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
| + | |
− | 1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
| + | |
− | 1801 ctcattaggc accccaggct ttacccgaac gaccgagcgc agcgagtcag tgagcgagga
| + | |
− | 1861 agcggagagc gcccaatacg caaggaaaca gctatgacca tgttaatgca gctggcacga
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− | 1921 caggtttccc gactggaaag cgggcagtga gcggaaggcc catgaggcca gttaattaag
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− | 1981 aggtaccgaa ttcgcggccg cttctagatg gccggccatc atcatcatca tcataccggc
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− | 2041 gacgtgtacc acgacggcgc ctgccccgaa gtgaagcccg tggacaactt cgactggtcc
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− | 2101 cagtaccacg gcaagtggtg ggaggtggcc aagtacccca gccccaacgg caagtatggc
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− | 2161 aagtgcggct gggccgagta cacccccgag ggcaagagcg tgaaggtgtc cagatacgac
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− | 2221 gtgatccacg gcaaagaata cttcatggaa ggcaccgcct accccgtggg cgacagcaag
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− | 2281 atcggcaaga tctaccacag ccggaccgtg ggcggctaca ccagaaagac cgtgttcaac
| + | |
− | 2341 gtgctgtcca ccgacaacaa gaactacatc atcggctaca gctgccgcta cgacgaggac
| + | |
− | 2401 aagaagggcc actgggacca cgtgtgggtg ctgtcccggt ccatggtgct gaccggcgag
| + | |
− | 2461 gccaagaccg ccgtggagaa ctacctgatc ggcagccccg tggtggacag ccagaaactg
| + | |
− | 2521 gtgtacagcg acttcagcga ggccgcctgc aaagtgaaca acaccggccg accagcctgt
| + | |
− | 2581 aagattccaa atgacctgaa gcagaaagtt atgaatcaca ccggcaaatc tgaactggag
| + | |
− | 2641 gagaaaaaat ccgagctgcg ccacaaactg aaatatgtgc ctcacgagta tatcgaactg
| + | |
− | 2701 atcgagatcg cccgtaatag tacccaagac cgtatcctgg aaatgaaagt gatggagttc
| + | |
− | 2761 ttcatgaaag tctatggcta tcgtggcaaa catctgggtg gtagccgtaa accagcaggt
| + | |
− | 2821 gccatttata ccgttggttc cccgatcgat tatggcgtta tcgtggccac aaaagcgtat
| + | |
− | 2881 tctggcggtt ataatctgcc gattggtcag gctgatgaga tggaacgtta tgtggaagag
| + | |
− | 2941 aatcagaccc gtaacaaaca tctgaacccg aacgaatggt ggaaagtgta tccgtcaagt
| + | |
− | 3001 gtcaccgagt tcaaatttct gttcgtgagc ggccacttta aaggcaacta taaagcccag
| + | |
− | 3061 ctgactcgtc tgaaccatat caccaatagc aatggggcag tgctgagtgt tgaggaactg
| + | |
− | 3121 ctgatcggtg gagaaatgat caaagcaggc accctgactc tggaagaagt tcgccgtaaa
| + | |
− | 3181 ttcaacaatg gcgagatcaa ttttaccggt taatactagt agcggccgct gcagagctca
| + | |
− | 3241 tggcgcgcct aggccttgac ggccttccgc caattcgccc tatagtgagt cgtattacgt
| + | |
− | 3301 cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
| + | |
− | 3361 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
| + | |
− | 3421 ccgaaacgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggagcg ccctgtagcg
| + | |
− | 3481 gccactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
| + | |
− | 3541 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
| + | |
− | 3601 aacgcttaca atttag
| + | |
− | //
| + | |
| | | |
| ===References=== | | ===References=== |
| + | Holtke HJ, Ankenbauer W, Muhlegger K, Rein R, Sagner G, Seibl R, Walter T. (1995)<br> |
| + | ''The digoxigenin (DIG) system for non-radioactive labelling and detection of nucleic |
| + | acids--an overview.'' Cell Mol Biol. 41(7):883-905.<br> |
| + | |
| + | Roche Applied Science (2002) ''DIG Application Manual for Nonradioactive In Situ |
| + | Hybridization'' (3rdedition) <br> |
| + | |
| + | Moter A, Göbel UB (2000) ''Fluorescence in situ hybridisation (FISH) for direct |
| + | visualization of microorganism'', J Microbiol Meth. 41: 85-112 |
| + | |
| + | http://www.thermo.com/eThermo/CMA/PDFs/Various/File_29495.pdf (image) |
Planned and designed by Team Freiburg Bioware09 and synthesized by purimex.
Holtke HJ, Ankenbauer W, Muhlegger K, Rein R, Sagner G, Seibl R, Walter T. (1995)
The digoxigenin (DIG) system for non-radioactive labelling and detection of nucleic
acids--an overview. Cell Mol Biol. 41(7):883-905.