LOCUS Exported 6379 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS GFP-cortactin SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6379) AUTHORS Perrin BJ, Amann KJ, Huttenlocher A TITLE Proteolysis of cortactin by calpain regulates membrane protrusion during cell migration. JOURNAL Mol Biol Cell. 2006 Jan . 17(1):239-50. PUBMED 16280362 REFERENCE 2 (bases 1 to 6379) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6379 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 52..355 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 356..559 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 510..530 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 598..607 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 604..1320 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 604..1320 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(649..670) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(910..929) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1257..1278 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 3158..3279 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3195..3214) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3249..3268 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3286..3741) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3423..3444) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3635..3654 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3768..3872 /gene="bla" /label=AmpR promoter promoter 3874..4231 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3897..3917) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4082..4217 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4144..4163 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4266..5060 /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(4320..4339) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4930..4949 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5248..5267) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5292..5339 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5668..6256 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6157..6176 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata 61 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat 121 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga 181 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc 241 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt 301 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat 361 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag 421 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc 481 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga 541 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggtcgcc 601 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg 661 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc 721 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc 781 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg 841 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc 901 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc 961 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg 1021 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag 1081 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc 1141 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac 1201 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg 1261 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag 1321 tccggactca gatctcgagc tcaagcttcg aattctatgt ggaaagcctc tgcaggccat 1381 gctgtgtcca tcacgcagga tgatggagga gctgatgact gggagactga tcctgatttt 1441 gtgaatgatg tgagtgaaaa ggagcagaga tggggtgcta aaaccgtgca gggatcgggg 1501 caccaggaac acatcaacat tcacaagctt cgagagaatg tcttccaaga acaccagacg 1561 ctcaaggaga aggagctgga aacgggaccc aaggcttccc acggctatgg cgggaagttc 1621 ggtgtggagc aggataggat ggacagatca gccgtgggcc atgagtacca gtcgaagctt 1681 tccaagcact gctcacaagt ggactcggtc cggggcttcg gaggcaagtt cggtgtccag 1741 atggacaggg tggatcagtc tgctgtaggc tttgaatacc aggggaagac tgagaagcat 1801 gcctcccaga aagactactc tagtggcttc ggtggcaaat acggtgtgca agctgaccgt 1861 gtagacaaga gtgccgtggg ctttgactac cagggcaaga cggagaagca tgagtctcag 1921 aaagattact ccaaaggttt tggtggcaaa tatgggattg acaaggacaa ggtggataaa 1981 agtgctgtgg gctttgagta tcaaggcaag acagagaagc acgaatccca gaaagactat 2041 gtaaaaggct ttggaggaaa gtttggtgtg cagacagaca gacaggacaa gtgtgccctt 2101 ggctgggacc atcaggagaa gctgcagctg catgaatccc aaaaagacta taagactggt 2161 ttcggaggca aatttggtgt tcagtccgag aggcaggact cctccgctgt ggggtttgat 2221 tacaaggaga gattggccaa gcacgagccc cagcaagact atgccaaagg attcggcggg 2281 aagtatgggg tgcagaagga tcggatggac aagaatgcat ccacctttga agaagtggtc 2341 caggtgccat ctgcctatca gaagactgtc cccattgagg ccgtaaccag caaaaccagt 2401 aatatccgtg ctaactttga aaacctggca aaggagagag agcaggagga caggcggaag 2461 gcagaagccg agagagctca gcggatggcc aaagaaagac aggagcagga ggaggcgcgc 2521 aggaagctgg aagagcaagc cagagccaag aagcagacgc cccctgcatc ccctagtcct 2581 caaccaattg aagacagacc accctccagc cccatctatg aggatgcagc tccgttcaag 2641 gccgagccga gctaccgagg tagcgaacct gagcctgagt acagcatcga ggccgcaggc 2701 attcctgagg ctggcagcca gcaaggcctg acctatacat cagagcccgt gtacgagact 2761 acagaggctc ctggccacta tcaagcagag gatgacacct acgatgggta tgagagtgac 2821 ctgggcatca cagccatcgc cctgtatgac taccaggctg ctggcgatga tgagatctcc 2881 tttgaccctg atgacatcat caccaacata gaaatgattg acgatggctg gtggcgtggg 2941 gtgtgcaagg gcagatacgg gctcttccca gccaactatg tggagctgcg gcagtaggaa 3001 ttctgcagtc gacggtaccg cgggcccggg atccaccgga tctagataac tgatcataat 3061 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct 3121 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa 3181 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 3241 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta 3301 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg 3361 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg 3421 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa 3481 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg 3541 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt 3601 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg 3661 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta 3721 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta 3781 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat 3841 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa 3901 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 3961 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag 4021 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc 4081 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt 4141 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg 4201 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt 4261 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc 4321 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc 4381 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg 4441 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag 4501 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg 4561 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg 4621 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac 4681 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg 4741 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc 4801 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg 4861 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc 4921 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc 4981 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc 5041 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc 5101 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg 5161 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt 5221 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac 5281 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca 5341 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg 5401 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg 5461 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata 5521 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct 5581 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 5641 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 5701 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 5761 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct 5821 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 5881 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 5941 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 6001 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 6061 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 6121 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 6181 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 6241 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 6301 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac 6361 cgccatgcat tagttatta //