LOCUS Exported 5141 bp ds-DNA circular SYN 15-JUN-2022 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pEGFP VAMP2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5141) AUTHORS Martinez-Arca S, Rudge R, Vacca M, Raposo G, Camonis J, Proux-Gillardeaux V, Daviet L, Formstecher E, Hamburger A, Filippini F, D'Esposito M, Galli T TITLE A dual mechanism controlling the localization and function of exocytic v-SNAREs. JOURNAL Proc Natl Acad Sci U S A. 2003 Jul 22;100(15):9011-6. Epub 2003 Jul 9. PUBMED 12853575 REFERENCE 2 (bases 1 to 5141) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 15, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5141 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 607..616 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 613..1329 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 613..1329 /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(658..679) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(919..938) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1266..1287 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 1929..2050 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1966..1985) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2020..2039 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2057..2512) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2194..2215) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2406..2425 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2539..2643 /gene="bla" /label=AmpR promoter promoter 2645..3002 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2668..2688) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2853..2988 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2915..2934 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3037..3831 /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(3091..3110) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3701..3720 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4019..4038) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4063..4110 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4439..5027 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4928..4947 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc 721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc 841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac 1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc 1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc 1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 1321 ctgtacaagt actcagatct cgagctcaag cttcgaattc tgcagtcgac ggtaccgagc 1381 tcggatccac tagtccagtg tggtggaatt gcccttatgt cggctaccgc tgccaccgcc 1441 ccccctgccg ccccggctgg ggagggaggc cctcctgcgc cccctccaaa cctcactagt 1501 aacaggagac tgcagcagac ccaggcccag gtggatgagg tggtggacat catgagggtg 1561 aacgtagaca aggtcttgga gcgggaccag aagctgtcgg agctggacga ccgtgccgac 1621 gccctccagg cgggggcctc ccagtttgaa actagtgcag ccaagctcaa gcgcaaatac 1681 tggtggaaaa acctcaagat gatgatcatc ctgggagtga tttgcgccat catcctcatt 1741 atcatcatag tttacttcag ctcttaaaag ggcaattctg cagatatcca gcacagtggc 1801 ggccgctcga gtctagataa ctgatcataa tcagccatac cacatttgta gaggttttac 1861 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg 1921 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 1981 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 2041 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt 2101 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca 2161 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta 2221 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta 2281 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg 2341 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga 2401 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg 2461 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc 2521 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat 2581 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag 2641 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc 2701 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt 2761 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 2821 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 2881 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 2941 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca 3001 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac 3061 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca 3121 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt 3181 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg 3241 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga 3301 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct 3361 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg 3421 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg 3481 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc 3541 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat 3601 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac 3661 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt 3721 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct 3781 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc 3841 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca 3901 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga 3961 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg 4021 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga 4081 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc 4141 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt 4201 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg 4261 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt 4321 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt 4381 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt 4441 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag 4501 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca 4561 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca 4621 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg 4681 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg 4741 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct 4801 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga 4861 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc 4921 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg 4981 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg 5041 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt 5101 tatcccctga ttctgtggat aaccgtatta ccgccatgca t //