LOCUS       sequence-65170-e        8659 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 8659)
  AUTHORS   Chen H, Cohen DM, Choudhury DM, Kioka N, Craig SW
  TITLE     Spatial distribution and functional significance of activated 
            vinculin in living cells.
  JOURNAL   J Cell Biol. 2005 May 9;169(3):459-70.
  PUBMED    15883197
REFERENCE   2  (bases 1 to 8659)
  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 Cell 
            Biol."; date: "2005-05-9"; volume: "169(3)"; pages: "459-70"
COMMENT     Sequence Label: GgVcl 1-1066 FRET tail probe 1
FEATURES             Location/Qualifiers
     source          1..8659
                     /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"
     CDS             join(3318..3320,3321..3323,3324..4034)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4605..4607,4608..4610,4611..5324)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    5447..5568
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5484..5503)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5538..5557
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5575..6030)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5712..5733)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5924..5943
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6057..6161
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6163..6520
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6163..6354
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6186..6206)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6371..6506
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6555..7349
                     /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             6555..7349
                     /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(6609..6628)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7219..7238
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7537..7556)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7581..7628
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7957..8545
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8446..8465
                     /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 ccggactcag atctcgagct caagcttcga attccggccc cccctgcccc gccgccgcca
      661 tgcccgtctt ccacacgcgc accatcgaga gcatcttgga gcccgtggct cagcagatct
      721 cccacctggt catcatgcac gaggaggggg aggtagacgg caaggccatc ccggacctca
      781 ccgcccccgt gtcggccgtg caggccgctg tcagcaacct ggtgcgggtt ggaaaagaaa
      841 ctgtgcagac aacagaagac cagatcttga aaagggatat gccaccagca ttcatcaaag
      901 tagagaatgc ctgcaccaag ctcgttcgag cagcccagat gctgcaagca gatccttatt
      961 cagtaccagc tcgtgactac ctaattgatg gatcaagagg catcctttct ggaacatcag
     1021 acttacttct gacatttgat gaagcggagg tccgtaaaat catccgtgtc tgcaaaggaa
     1081 tattggaata tctgactgtg gcagaagtag tagagactat ggaggatttg gtgacatata
     1141 caaagaatct agggccagga atgacaaaga tggcgaaaat gatcgatgag agacaacagg
     1201 aattaactca tcaggaacat agggttatgc tggtgaactc catgaatact gtgaaggagc
     1261 tattgcctgt acttatttca gctatgaaga tctttgtaac cacaaaaaac actaaaagcc
     1321 agggaataga agaggccttg aaaaatcgca atttcacagt agagaaaatg agtgctgaga
     1381 taaatgaaat aatccgtgta ttacaactca cttcctggga tgaagatgcc tgggccagca
     1441 aggacactga agccatgaaa agagctttag ccctaataga ttcaaagatg aaccaggcaa
     1501 aaggctggct gagagatcca aacgcacctc caggggatgc tggtgagcaa gcaataaggc
     1561 agatccttga tgaagctgga aaagcaggag aattgtgtgc aggcaaagaa cgcagggaga
     1621 ttctgggaac atgcaaaact ttgggccaaa tgactgatca acttgctgat ctccgagcta
     1681 gaggacaggg tgctacaccg atggcaatgc agaaggcaca gcaggtgtca caaggcctgg
     1741 atttgctcac tgcaaaagtg gagaatgcag cccggaaatt ggaggccatg acaaactcta
     1801 agcaggctat tgcaaagaag attgatgctg ctcagaactg gcttgcggat cccaacggcg
     1861 gaagtgaagg agaagaacac attcgaggaa ttatgtctga agcaaggaaa gttgcagaat
     1921 tgtgtgagga gcctaaagaa agagatgata tccttcgctc cttgggggaa atctctgctc
     1981 tgacagctaa gctgtcagat ctgcgacgac atgggaaagg cgactctcct gaggcccgtg
     2041 cattggccaa gcaaatagct acatcacttc agaacttaca gtccaaaaca aacagagctg
     2101 tagcaaatac taggccagtt aaagctgctg tccatttgga gggcaagatt gagcaagccc
     2161 agaggtggat agataatcct acagttgatg atcggggagt aggccaggca gcaattcggg
     2221 gtttggttgc agaaggtcgt cgtctagcca atgtcatgat gggaccttat cgtcaggacc
     2281 tgcttgccaa atgtgaccgt gtagaccagc tggctgctca gcttgctgac cttgcagcaa
     2341 gaggggaagg agagtctcct caggctaggg caattgctgc tcagcttcag gactccctga
     2401 aggatctcaa agcacggatg caagaagcaa tgacccagga ggtgtctgat gttttcagtg
     2461 acactacaac ccctattaag ttgttagcag tagcagccac tgctccttct gatactccca
     2521 atagagaaga ggtgtttgaa gaaagagcag caaattttga aaaccatgct gctagactgg
     2581 gagcaacagc agaaaaagca gctgcagttg gaactgctaa taaaactact gtggaaggca
     2641 ttcaggcaac agtcaaatct gcaagggagc ttacaccaca ggtagtatcg gctgctcgaa
     2701 tcctcctgag aaatcctgga aatcaagctg cttatgagca ttttgagaca atgaaaaacc
     2761 aatggattga taatgtagaa aagatgacag ggctggtgga tgaggccatc gataccaagt
     2821 ctctgttgga tgcatcagaa gaggctatta agaaggatct tgataaatgc aaagttgcaa
     2881 tggccaatat gcaacctcag atgctggtag ctggagccac cagcattgct agacgagcga
     2941 accgcatcct gcttgtggca aaacgggagg ttgaaaattc agaagaccct aaattcaggg
     3001 aggctgttaa agcagcttct gatgagctga gcaaaaccat atcaccgatg gtaatggatg
     3061 ctaaagctgt agcaggaaat atctctgatc ctggtttgca gaagagtttc ttggattctg
     3121 gatacaggat tctgggagct gtggccaaag tcagagaagc ctttcagcct caggagccag
     3181 actttccccc tcctcctcct gacctagagc atctccatct gactgatgag cttgctcctc
     3241 caaaaccacc acttccagaa ggtgaggttc ccccacccag accaccaccc cctgaagaaa
     3301 aagatgaggg cggccgcatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     3361 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     3421 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     3481 ccgtgccctg gcccaccctc gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct
     3541 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     3601 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     3661 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     3721 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     3781 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     3841 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     3901 tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac cccaacgaga
     3961 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     4021 acgagctgta caagggcggc cgcgagttcc cagagcagaa agcaggagaa gctattaatc
     4081 agcccatgat gatggctgct aggcagttgc atgacgaggc tcggaaatgg tctagcaagg
     4141 gtaacgacat cattgctgct gctaaacgaa tggcgctgct catggctgag atgtcacgcc
     4201 tggtgcgagg aggcagcgga aacaagcgtg ccctcatcca gtgtgcaaaa gatattgcta
     4261 aggcatcgga tgaagtcact cgattggcca aagaggtggc caagcaatgt actgataaac
     4321 gcattagaac aaacctctta caggtctgtg agcgaatccc aaccatcagc acgcagctca
     4381 aaattctttc cacagtgaaa gctaccatgc tgggcaggac taacatcagc gatgaagaat
     4441 cagaacaggc aactgagatg ttggttcata acgcccagaa cctcatgcag tctgtgaagg
     4501 aaactgtgag agaagctgaa gcagcatcca ttaagataag aacagatgcc ggattcactc
     4561 tgcgctgggt cagaaagacc ccatggtatc aggtcgacgg taccatggtg agcaagggcg
     4621 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     4681 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     4741 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     4801 cctggggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     4861 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     4921 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     4981 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     5041 acatcagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     5101 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     5161 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     5221 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     5281 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta
     5341 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     5401 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     5461 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5521 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg
     5581 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     5641 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     5701 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     5761 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     5821 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     5881 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     5941 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     6001 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     6061 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     6121 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     6181 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     6241 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     6301 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     6361 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     6421 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     6481 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     6541 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     6601 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     6661 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     6721 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     6781 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     6841 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     6901 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     6961 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     7021 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     7081 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     7141 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     7201 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     7261 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     7321 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     7381 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     7441 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     7501 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     7561 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     7621 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     7681 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     7741 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     7801 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     7861 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     7921 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     7981 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     8041 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     8101 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     8161 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     8221 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     8281 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     8341 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     8401 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     8461 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     8521 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     8581 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     8641 ccgtattacc gccatgcat
//
