LOCUS       sequence-65141-e        7989 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 7989)
  AUTHORS   Cohen DM, Chen H, Johnson RP, Choudhury B, Craig SW
  TITLE     Two distinct head-tail interfaces cooperate to suppress activation 
            of vinculin by talin.
  JOURNAL   J Biol Chem. 2005 Apr 29;280(17):17109-17. Epub 2005 Feb 22.
  PUBMED    15728584
REFERENCE   2  (bases 1 to 7989)
  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: "2005-04-29"; volume: "280(17)"; pages: "17109-17. 
            Epub 2005 Feb 22"
COMMENT     Sequence Label: pEGFPC1/GgVcl 1-1066
FEATURES             Location/Qualifiers
     source          1..7989
                     /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    4777..4898
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4814..4833)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4868..4887
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4905..5360)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5042..5063)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5254..5273
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5387..5491
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5493..5850
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5493..5684
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5516..5536)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5701..5836
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5885..6679
                     /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             5885..6679
                     /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(5939..5958)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6549..6568
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6867..6886)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6911..6958
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7287..7875
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7776..7795
                     /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 ccggactcag atctcgagct caagcttcga attccggccc cccctgcccc
     1381 gccgccgcca tgcccgtctt ccacacgcgc accatcgaga gcatcttgga gcccgtggct
     1441 cagcagatct cccacctggt catcatgcac gaggaggggg aggtagacgg caaggccatc
     1501 ccggacctca ccgcccccgt gtcggccgtg caggccgctg tcagcaacct ggtgcgggtt
     1561 ggaaaagaaa ctgtgcagac aacagaagac cagatcttga aaagggatat gccaccagca
     1621 ttcatcaaag tagagaatgc ctgcaccaag ctcgttcgag cagcccagat gctgcaagca
     1681 gatccttatt cagtaccagc tcgtgactac ctaattgatg gatcaagagg catcctttct
     1741 ggaacatcag acttacttct gacatttgat gaagcggagg tccgtaaaat catccgtgtc
     1801 tgcaaaggaa tattggaata tctgactgtg gcagaagtag tagagactat ggaggatttg
     1861 gtgacatata caaagaatct agggccagga atgacaaaga tggcgaaaat gatcgatgag
     1921 agacaacagg aattaactca tcaggaacat agggttatgc tggtgaactc catgaatact
     1981 gtgaaggagc tattgcctgt acttatttca gctatgaaga tctttgtaac cacaaaaaac
     2041 actaaaagcc agggaataga agaggccttg aaaaatcgca atttcacagt agagaaaatg
     2101 agtgctgaga taaatgaaat aatccgtgta ttacaactca cttcctggga tgaagatgcc
     2161 tgggccagca aggacactga agccatgaaa agagctttag ccctaataga ttcaaagatg
     2221 aaccaggcaa aaggctggct gagagatcca aacgcacctc caggggatgc tggtgagcaa
     2281 gcaataaggc agatccttga tgaagctgga aaagcaggag aattgtgtgc aggcaaagaa
     2341 cgcagggaga ttctgggaac atgcaaaact ttgggccaaa tgactgatca acttgctgat
     2401 ctccgagcta gaggacaggg tgctacaccg atggcaatgc agaaggcaca gcaggtgtca
     2461 caaggcctgg atttgctcac tgcaaaagtg gagaatgcag cccggaaatt ggaggccatg
     2521 acaaactcta agcaggctat tgcaaagaag attgatgctg ctcagaactg gcttgcggat
     2581 cccaacggcg gaagtgaagg agaagaacac attcgaggaa ttatgtctga agcaaggaaa
     2641 gttgcagaat tgtgtgagga gcctaaagaa agagatgata tccttcgctc cttgggggaa
     2701 atctctgctc tgacagctaa gctgtcagat ctgcgacgac atgggaaagg cgactctcct
     2761 gaggcccgtg cattggccaa gcaaatagct acatcacttc agaacttaca gtccaaaaca
     2821 aacagagctg tagcaaatac taggccagtt aaagctgctg tccatttgga gggcaagatt
     2881 gagcaagccc agaggtggat agataatcct acagttgatg atcggggagt aggccaggca
     2941 gcaattcggg gtttggttgc agaaggtcgt cgtctagcca atgtcatgat gggaccttat
     3001 cgtcaggacc tgcttgccaa atgtgaccgt gtagaccagc tggctgctca gcttgctgac
     3061 cttgcagcaa gaggggaagg agagtctcct caggctaggg caattgctgc tcagcttcag
     3121 gactccctga aggatctcaa agcacggatg caagaagcaa tgacccagga ggtgtctgat
     3181 gttttcagtg acactacaac ccctattaag ttgttagcag tagcagccac tgctccttct
     3241 gatactccca atagagaaga ggtgtttgaa gaaagagcag caaattttga aaaccatgct
     3301 gctagactgg gagcaacagc agaaaaagca gctgcagttg gaactgctaa taaaactact
     3361 gtggaaggca ttcaggcaac agtcaaatct gcaagggagc ttacaccaca ggtagtatcg
     3421 gctgctcgaa tcctcctgag aaatcctgga aatcaagctg cttatgagca ttttgagaca
     3481 atgaaaaacc aatggattga taatgtagaa aagatgacag ggctggtgga tgaggccatc
     3541 gataccaagt ctctgttgga tgcatcagaa gaggctatta agaaggatct tgataaatgc
     3601 aaagttgcaa tggccaatat gcaacctcag atgctggtag ctggagccac cagcattgct
     3661 agacgagcga accgcatcct gcttgtggca aaacgggagg ttgaaaattc agaagaccct
     3721 aaattcaggg aggctgttaa agcagcttct gatgagctga gcaaaaccat atcaccgatg
     3781 gtaatggatg ctaaagctgt agcaggaaat atctctgatc ctggtttgca gaagagtttc
     3841 ttggattctg gatacaggat tctgggagct gtggccaaag tcagagaagc ctttcagcct
     3901 caggagccag actttccccc tcctcctcct gacctagagc atctccatct gactgatgag
     3961 cttgctcctc caaaaccacc acttccagaa ggtgaggttc ccccacccag accaccaccc
     4021 cctgaagaaa aagatgagga gttcccagag cagaaagcag gagaagctat taatcagccc
     4081 atgatgatgg ctgctaggca gttgcatgac gaggctcgga aatggtctag caagggtaac
     4141 gacatcattg ctgctgctaa acgaatggcg ctgctcatgg ctgagatgtc acgcctggtg
     4201 cgaggaggca gcggaaacaa gcgtgccctc atccagtgtg caaaagatat tgctaaggca
     4261 tcggatgaag tcactcgatt ggccaaagag gtggccaagc aatgtactga taaacgcatt
     4321 agaacaaacc tcttacaggt ctgtgagcga atcccaacca tcagcacgca gctcaaaatt
     4381 ctttccacag tgaaagctac catgctgggc aggactaaca tcagcgatga agaatcagaa
     4441 caggcaactg agatgttggt tcataacgcc cagaacctca tgcagtctgt gaaggaaact
     4501 gtgagagaag ctgaagcagc atccattaag ataagaacag atgccggatt cactctgcgc
     4561 tgggtcagaa agaccccatg gtatcagtaa acaccacata agcattgtct tctgtcgaat
     4621 tctcgagtcg acggtaccgc gggcccggga tccaccggat ctagataact gatcataatc
     4681 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     4741 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     4801 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4861 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa
     4921 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     4981 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     5041 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     5101 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     5161 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     5221 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     5281 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     5341 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5401 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5461 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     5521 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5581 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     5641 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     5701 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5761 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     5821 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     5881 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     5941 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     6001 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     6061 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     6121 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     6181 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     6241 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     6301 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     6361 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     6421 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     6481 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     6541 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     6601 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     6661 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     6721 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     6781 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     6841 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     6901 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     6961 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     7021 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     7081 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     7141 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     7201 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     7261 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     7321 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     7381 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     7441 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     7501 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     7561 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     7621 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     7681 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7741 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7801 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     7861 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     7921 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     7981 gccatgcat
//
