LOCUS       sequence-65190-e        7924 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 7924)
  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 7924)
  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: pmECFPN3/GgVcl 1-1066 (CFP at C term)
FEATURES             Location/Qualifiers
     source          1..7924
                     /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(3870..3872,3873..3875,3876..4589)
                     /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(3915..3936)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4176..4195)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4523..4544
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4712..4833
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4749..4768)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4803..4822
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4840..5295)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4977..4998)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5189..5208
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5322..5426
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5428..5785
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5428..5619
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5451..5471)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5636..5771
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5820..6614
                     /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             5820..6614
                     /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(5874..5893)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6484..6503
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6802..6821)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6846..6893
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7222..7810
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7711..7730
                     /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 aagatgagga gttcccagag cagaaagcag gagaagctat taatcagccc atgatgatgg
     3361 ctgctaggca gttgcatgac gaggctcgga aatggtctag caagggtaac gacatcattg
     3421 ctgctgctaa acgaatggcg ctgctcatgg ctgagatgtc acgcctggtg cgaggaggca
     3481 gcggaaacaa gcgtgccctc atccagtgtg caaaagatat tgctaaggca tcggatgaag
     3541 tcactcgatt ggccaaagag gtggccaagc aatgtactga taaacgcatt agaacaaacc
     3601 tcttacaggt ctgtgagcga atcccaacca tcagcacgca gctcaaaatt ctttccacag
     3661 tgaaagctac catgctgggc aggactaaca tcagcgatga agaatcagaa caggcaactg
     3721 agatgttggt tcataacgcc cagaacctca tgcagtctgt gaaggaaact gtgagagaag
     3781 ctgaagcagc atccattaag ataagaacag atgccggatt cactctgcgc tgggtcagaa
     3841 agaccccatg gtatcaggtc gacggtacca tggtgagcaa gggcgaggag ctgttcaccg
     3901 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     3961 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     4021 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctgg ggcgtgcagt
     4081 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     4141 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     4201 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     4261 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacatc agccacaacg
     4321 tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca
     4381 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     4441 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaaa ctgagcaaag
     4501 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     4561 ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca taatcagcca
     4621 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     4681 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     4741 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4801 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     4861 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     4921 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     4981 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     5041 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     5101 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     5161 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     5221 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     5281 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5341 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5401 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     5461 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5521 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5581 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     5641 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5701 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     5761 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     5821 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5881 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     5941 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     6001 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     6061 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     6121 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     6181 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     6241 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     6301 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     6361 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     6421 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     6481 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     6541 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     6601 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     6661 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     6721 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     6781 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     6841 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     6901 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     6961 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     7021 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     7081 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     7141 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7201 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7261 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     7321 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     7381 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7441 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7501 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7561 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     7621 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7681 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7741 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7801 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7861 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     7921 gcat
//
