LOCUS       sequence-65193-e        7950 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 7950)
  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 7950)
  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: pmECFPC1/GgVcl 1-1066 (CFP at N term)
FEATURES             Location/Qualifiers
     source          1..7950
                     /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 CFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     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    4738..4859
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4775..4794)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4829..4848
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4866..5321)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5003..5024)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5215..5234
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5348..5452
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5454..5811
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5454..5645
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5477..5497)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5662..5797
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5846..6640
                     /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             5846..6640
                     /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(5900..5919)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6510..6529
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6828..6847)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6872..6919
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7248..7836
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7737..7756
                     /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 tggggcgtgc 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 atcagccaca acgtctatat caccgccgac
     1081 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc aaactgagca 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 gtatcaggtc gacggtaccg cgggcccggg atccaccgga
     4621 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     4681 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     4741 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4801 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     4861 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     4921 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     4981 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     5041 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     5101 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     5161 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     5221 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     5281 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     5341 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5401 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     5461 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     5521 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     5581 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     5641 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5701 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     5761 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     5821 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     5881 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     5941 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     6001 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     6061 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     6121 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     6181 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     6241 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     6301 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     6361 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     6421 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     6481 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     6541 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     6601 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     6661 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     6721 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     6781 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     6841 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     6901 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     6961 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     7021 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     7081 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     7141 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     7201 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     7261 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7321 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7381 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7441 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7501 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7561 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7621 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7681 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7741 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7801 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7861 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7921 tctgtggata accgtattac cgccatgcat
//
