LOCUS       sequence-65156-e        7316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7316)
  AUTHORS   Cohen DM, Kutscher B, Chen H, Murphy DB, Craig SW
  TITLE     A conformational switch in vinculin drives formation and dynamics of
            a talin-vinculin complex at focal adhesions.
  JOURNAL   J Biol Chem. 2006 Jun 9;281(23):16006-15. Epub 2006 Apr 10.
  PUBMED    16608855
REFERENCE   2  (bases 1 to 7316)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem."; date: "2006-06-9"; volume: "281(23)"; pages: "16006-15. Epub
            2006 Apr 10"
COMMENT     Sequence Label: pEGFPC1/GgVcl 1-851 A50I mutant
FEATURES             Location/Qualifiers
     source          1..7316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..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    4104..4225
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4141..4160)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4195..4214
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4232..4687)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4369..4390)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4581..4600
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4714..4818
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4820..5177
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4820..5011
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4843..4863)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5028..5163
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5212..6006
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5212..6006
                     /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(5266..5285)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5876..5895
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6194..6213)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6238..6285
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6614..7202
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7103..7122
                     /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 atcttatgcc
      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 ccggactcag atctagagct caagcttcga attccggccc cccctgcccc
     1381 gccgcccata tgatgcccgt cttccacacg cgcaccatcg agagcatctt ggagcccgtg
     1441 gctcagcaga tctcccacct ggtcatcatg cacgaggagg gggaggtaga cggcaaggcc
     1501 atcccggacc tcaccgcccc cgtgtcggcc gtgcaggcca tcgtcagcaa cctggtgcgg
     1561 gttggaaaag aaactgtgca gacaacagaa gaccagatct tgaaaaggga tatgccacca
     1621 gcattcatca aagtagagaa tgcctgcacc aagctcgttc gagcagccca gatgctgcaa
     1681 gcagatcctt attcagtacc agctcgtgac tacctaattg atggatcaag aggcatcctt
     1741 tctggaacat cagacttact tctgacattt gatgaagcgg aggtccgtaa aatcatccgt
     1801 gtctgcaaag gaatattgga atatctgact gtggcagaag tagtagagac tatggaggat
     1861 ttggtgacat atacaaagaa tctagggcca ggaatgacaa agatggcgaa aatgatcgat
     1921 gagagacaac aggaattaac tcatcaggaa catagggtta tgctggtgaa ctccatgaat
     1981 actgtgaagg agctattgcc tgtacttatt tcagctatga agatctttgt aaccacaaaa
     2041 aacactaaaa gccagggaat agaagaggcc ttgaaaaatc gcaatttcac agtagagaaa
     2101 atgagtgctg agataaatga aataatccgt gtattacaac tcacttcctg ggatgaagat
     2161 gcctgggcca gcaaggacac tgaagccatg aaaagagctt tagccctaat agattcaaag
     2221 atgaaccagg caaaaggctg gctgagagat ccaaacgcac ctccagggga tgctggtgag
     2281 caagcaataa ggcagatcct tgatgaagct ggaaaagcag gagaattgtg tgcaggcaaa
     2341 gaacgcaggg agattctggg aacatgcaaa actttgggcc aaatgactga tcaacttgct
     2401 gatctccgag ctagaggaca gggtgctaca ccgatggcaa tgcagaaggc acagcaggtg
     2461 tcacaaggcc tggatttgct cactgcaaaa gtggagaatg cagcccggaa attggaggcc
     2521 atgacaaact ctaagcaggc tattgcaaag aagattgatg ctgctcagaa ctggcttgcg
     2581 gatcccaacg gcggaagtga aggagaagaa cacattcgag gaattatgtc tgaagcaagg
     2641 aaagttgcag aattgtgtga ggagcctaaa gaaagagatg atatccttcg ctccttgggg
     2701 gaaatctctg ctctgacagc taagctgtca gatctgcgac gacatgggaa aggcgactct
     2761 cctgaggccc gtgcattggc caagcaaata gctacatcac ttcagaactt acagtccaaa
     2821 acaaacagag ctgtagcaaa tactaggcca gttaaagctg ctgtccattt ggagggcaag
     2881 attgagcaag cccagaggtg gatagataat cctacagttg atgatcgggg agtaggccag
     2941 gcagcaattc ggggtttggt tgcagaaggt cgtcgtctag ccaatgtcat gatgggacct
     3001 tatcgtcagg acctgcttgc caaatgtgac cgtgtagacc agctggctgc tcagcttgct
     3061 gaccttgcag caagagggga aggagagtct cctcaggcta gggcaattgc tgctcagctt
     3121 caggactccc tgaaggatct caaagcacgg atgcaagaag caatgaccca ggaggtgtct
     3181 gatgttttca gtgacactac aacccctatt aagttgttag cagtagcagc cactgctcct
     3241 tctgatactc ccaatagaga agaggtgttt gaagaaagag cagcaaattt tgaaaaccat
     3301 gctgctagac tgggagcaac agcagaaaaa gcagctgcag ttggaactgc taataaaact
     3361 actgtggaag gcattcaggc aacagtcaaa tctgcaaggg agcttacacc acaggtagta
     3421 tcggctgctc gaatcctcct gagaaatcct ggaaatcaag ctgcttatga gcattttgag
     3481 acaatgaaaa accaatggat tgataatgta gaaaagatga cagggctggt ggatgaggcc
     3541 atcgatacca agtctctgtt ggatgcatca gaagaggcta ttaagaagga tcttgataaa
     3601 tgcaaagttg caatggccaa tatgcaacct cagatgctgg tagctggagc caccagcatt
     3661 gctagacgag cgaaccgcat cctgcttgtg gcaaaacggg aggttgaaaa ttcagaagac
     3721 cctaaattca gggaggctgt taaagcagct tctgatgagc tgagcaaaac catatcaccg
     3781 atggtaatgg atgctaaagc tgtagcagga aatatctctg atcctggttt gcagaagagt
     3841 ttcttggatt ctggatacag gattctggga gctgtggcca aagtcagaga agcctttcag
     3901 cctcaggagc cagactttcc ccctcctcct cctgacctag agcattaact cgagtcgacg
     3961 gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc cataccacat
     4021 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     4081 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     4141 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4201 tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa
     4261 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     4321 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     4381 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     4441 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     4501 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     4561 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     4621 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     4681 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4741 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4801 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     4861 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4921 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4981 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     5041 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     5101 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5161 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     5221 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     5281 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     5341 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     5401 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     5461 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5521 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5581 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5641 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5701 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     5761 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5821 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5881 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5941 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     6001 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     6061 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     6121 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     6181 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     6241 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     6301 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     6361 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     6421 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     6481 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6541 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6601 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6661 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6721 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     6781 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6841 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6901 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6961 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     7021 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     7081 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     7141 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     7201 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     7261 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
