LOCUS Exported 7933 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Alpha 5 integrin-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7933) AUTHORS Laukaitis CM, Webb DJ, Donais K, Horwitz AF TITLE Differential dynamics of alpha 5 integrin, paxillin, and alpha-actinin during formation and disassembly of adhesions in migrating cells. JOURNAL J Cell Biol. 2001 Jun 25. 153(7):1427-40. PUBMED 11425873 REFERENCE 2 (bases 1 to 7933) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7933 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 119..422 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 423..626 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 577..597 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 3931..3940 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3937..4656 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3937..4653 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3982..4003) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4243..4262) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4590..4611 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 4779..4900 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4816..4835) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4870..4889 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(4907..5362) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5044..5065) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5256..5275 /label=F1ori-R /note="F1 origin, reverse primer" promoter 5389..5493 /gene="bla" /label=AmpR promoter promoter 5495..5852 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(5518..5538) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 5703..5838 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5765..5784 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5887..6681 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5941..5960) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6551..6570 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(6869..6888) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 6913..6960 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 7289..7877 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7778..7797 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta 61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat 481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac 601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc 661 ggactcagat ctcgagctca agcttgcatg cctgcaggtc gacctgcagc ccaagctcga 721 tgaattcgag ctcggtaccc ggggatcctc tagaccgcta tggggagccg gacgccagag 781 tcccctctcc acgccgtgca gctgcgctgg ggcccccggc gccgaccccc gctgctgccg 841 ctgctgttgc tgctgctgcc gccgccaccc agggtcgggg gcttcaactt agacgcggag 901 gccccagcag tactctcggg gcccccgggc tccttcttcg gattctcagt ggagttttac 961 cggccgggaa cagacggggt cagtgtgctg gtgggagcac ccaaggctaa taccagccag 1021 ccaggagtgc tgcagggtgg tgctgtctac ctctgtcctt ggggtgccag ccccacacag 1081 tgcaccccca ttgaatttga cagcaaaggc tctcggctcc tggagtcctc actgtccagc 1141 tcagagggag aggagcctgt ggagtacaag tccttgcagt ggttcggggc aacagttcga 1201 gcccatggct cctccatctt ggcatgcgct ccactgtaca gctggcgcac agagaaggag 1261 ccactgagcg accccgtggg cacctgctac ctctccacag ataacttcac ccgaattctg 1321 gagtatgcac cctgccgctc agatttcagc tgggcagcag gacagggtta ctgccaagga 1381 ggcttcagtg ccgagttcac caagactggc cgtgtggttt taggtggacc aggaagctat 1441 ttctggcaag gccagatcct gtctgccact caggagcaga ttgcagaatc ttattacccc 1501 gagtacctga tcaacctggt tcaggggcag ctgcagactc gccaggccag ttccatctat 1561 gatgacagct acctaggata ctctgtggct gttggtgaat tcagtggtga tgacacagaa 1621 gactttgttg ctggtgtgcc caaagggaac ctcacttacg gctatgtcac catccttaat 1681 ggctcagaca ttcgatccct ctacaacttc tcaggggaac agatggcctc ctactttggc 1741 tatgcagtgg ccgccacaga cgtcaatggg gacgggctgg atgacttgct ggtgggggca 1801 cccctgctca tggatcggac ccctgacggg cggcctcagg aggtgggcag ggtctacgtc 1861 tacctgcagc acccagccgg catagagccc acgcccaccc ttaccctcac tggccatgat 1921 gagtttggcc gatttggcag ctccttgacc cccctggggg acctggacca ggatggctac 1981 aatgatgtgg ccatcggggc tccctttggt ggggagaccc agcagggagt agtgtttgta 2041 tttcctgggg gcccaggagg gctgggctct aagccttccc aggttctgca gcccctgtgg 2101 gcagccagcc acaccccaga cttctttggc tctgcccttc gaggaggccg agacctggat 2161 ggcaatggat atcctgatct gattgtgggg tcctttggtg tggacaaggc tgtggtatac 2221 aggggccgcc ccatcgtgtc cgctagtgcc tccctcacca tcttccccgc catgttcaac 2281 ccagaggagc ggagctgcag cttagagggg aaccctgtgg cctgcatcaa ccttagcttc 2341 tgcctcaatg cttctggaaa acacgttgct gactccattg gtttcacagt ggaacttcag 2401 ctggactggc agaagcagaa gggaggggta cggcgggcac tgttcctggc ctccaggcag 2461 gcaaccctga cccagaccct gctcatccag aatggggctc gagaggattg cagagagatg 2521 aagatctacc tcaggaacga gtcagaattt cgagacaaac tctcgccgat tcacatcgct 2581 ctcaacttct ccttggaccc ccaagcccca gtggacagcc acggcctcag gccagcccta 2641 cattatcaga gcaagagccg gatagaggac aaggctcaga tcttgctgga ctgtggagaa 2701 gacaacatct gtgtgcctga cctgcagctg gaagtgtttg gggagcagaa ccatgtgtac 2761 ctgggtgaca agaatgccct gaacctcact ttccatgccc agaatgtggg tgagggtggc 2821 gcctatgagg ctgagcttcg ggtcaccgcc cctccagagg ctgagtactc aggactcgtc 2881 agacacccag ggaacttctc cagcctgagc tgtgactact ttgccgtgaa ccagagccgc 2941 ctgctggtgt gtgacctggg caaccccatg aaggcaggag ccagtctgtg gggtggcctt 3001 cggtttacag tccctcatct ccgggacact aagaaaacca tccagtttga cttccagatc 3061 ctcagcaaga atctcaacaa ctcgcaaagc gacgtggttt cctttcggct ctccgtggag 3121 gctcaggccc aggtcaccct gaacggtgtc tccaagcctg aggcagtgct attcccagta 3181 agcgactggc atccccgaga ccagcctcag aaggaggagg acctgggacc tgctgtccac 3241 catgtctatg agctcatcaa ccaaggcccc agctccatta gccagggtgt gctggaactc 3301 agctgtcccc aggctctgga aggtcagcag ctcctatatg tgaccagagt tacgggactc 3361 aactgcacca ccaatcaccc cattaaccca aagggcctgg agttggatcc cgagggttcc 3421 ctgcaccacc agcaaaaacg ggaagctcca agccgcagct ctgcttcctc gggacctcag 3481 atcctgaaat gcccggaggc tgagtgtttc aggctgcgct gtgagctcgg gcccctgcac 3541 caacaagaga gccaaagtct gcagttgcat ttccgagtct gggccaagac tttcttgcag 3601 cgggagcacc agccatttag cctgcagtgt gaggctgtgt acaaagccct gaagatgccc 3661 taccgaatcc tgcctcggca gctgccccaa aaagagcgtc aggtggccac agctgtgcaa 3721 tggaccaagg cagaaggcag ctatggcgtc ccactgtgga tcatcatcct agccatcctg 3781 tttggcctcc tgctcctagg tctactcatc tacatcctct acaagcttgg attcttcaaa 3841 cgctccctcc catatggcac cgccatggaa aaagctcagc tcaagcctcc agccacctct 3901 gatgccggta ccgcgggccc gggatccatc gccaccatgg tgagcaaggg cgaggagctg 3961 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc 4021 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc 4081 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc 4141 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc 4201 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag 4261 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc 4321 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc 4381 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc 4441 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc 4501 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg 4561 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc 4621 gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa 4681 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc 4741 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata 4801 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc 4861 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt 4921 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag 4981 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt 5041 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga 5101 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg 5161 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct 5221 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc 5281 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt 5341 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct 5401 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga 5461 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga 5521 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 5581 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca 5641 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 5701 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt 5761 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag 5821 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg 5881 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg 5941 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg 6001 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat 6061 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca 6121 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg 6181 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat 6241 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa 6301 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg 6361 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg 6421 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg 6481 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat 6541 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac 6601 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc 6661 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc 6721 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg 6781 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt 6841 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa 6901 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc 6961 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt 7021 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag 7081 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat 7141 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc 7201 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag 7261 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct 7321 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac 7381 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc 7441 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg 7501 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt 7561 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt 7621 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc 7681 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca 7741 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata 7801 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg 7861 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct 7921 ggccttttgc tca //