LOCUS       sequence-209871-        5880 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 5880)
  AUTHORS   Smyth JW, Vogan JM, Buch PJ, Zhang SS, Fong TS, Hong TT, Shaw RM
  TITLE     Actin cytoskeleton rest stops regulate anterograde traffic of 
            connexin 43 vesicles to the plasma membrane.
  JOURNAL   Circ Res. 2012 Mar 30;110(7):978-89. Epub 2012 Feb 9.
  PUBMED    22328533
REFERENCE   2  (bases 1 to 5880)
  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: "Circ Res.";
            date: "2012-03-30"; volume: "110(7)"; pages: "978-89. Epub 2012 Feb 
            9"
COMMENT     Sequence Label: pDEST/hCx43-EGFP-N1
FEATURES             Location/Qualifiers
     source          1..5880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    660..684
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1834..1858)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1878..1887
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1884..1886,1887..1889,1890..2603)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1884..1886,1887..1889,1890..2600)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1929..1950)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2190..2209)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2537..2558
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2726..2847
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2763..2782)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2817..2836
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2854..3309)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2991..3012)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3203..3222
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3336..3440
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3442..3799
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3442..3633
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3465..3485)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3650..3785
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3834..4628
                     /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             3834..4628
                     /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(3888..3907)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4498..4517
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4816..4835)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4860..4907
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5236..5824
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5725..5744
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcatcaaa
      661 caagtttgta caaaaaagca ggcttcatgg gtgactggag cgccttaggc aaactccttg
      721 acaaggttca agcctactca actgctggag ggaaggtgtg gctgtcagta cttttcattt
      781 tccgaatcct gctgctgggg acagcggttg agtcagcctg gggagatgag cagtctgcct
      841 ttcgttgtaa cactcagcaa cctggttgtg aaaatgtctg ctatgacaag tctttcccaa
      901 tctctcatgt gcgcttctgg gtcctgcaga tcatatttgt gtctgtaccc acactcttgt
      961 acctggctca tgtgttctat gtgatgcgaa aggaagagaa actgaacaag aaagaggaag
     1021 aactcaaggt tgcccaaact gatggtgtca atgtggacat gcacttgaag cagattgaga
     1081 taaagaagtt caagtacggt attgaagagc atggtaaggt gaaaatgcga ggggggttgc
     1141 tgcgaaccta catcatcagt atcctcttca agtctatctt tgaggtggcc ttcttgctga
     1201 tccagtggta catctatgga ttcagcttga gtgctgttta cacttgcaaa agagatccct
     1261 gcccacatca ggtggactgt ttcctctctc gccccacgga gaaaaccatc ttcatcatct
     1321 tcatgctggt ggtgtccttg gtgtccctgg ccttgaatat cattgaactc ttctatgttt
     1381 tcttcaaggg cgttaaggat cgggttaagg gaaagagcga cccttaccat gcgaccagtg
     1441 gtgcgctgag ccctgccaaa gactgtgggt ctcaaaaata tgcttatttc aatggctgct
     1501 cctcaccaac cgctcccctc tcgcctatgt ctcctcctgg gtacaagctg gttactggcg
     1561 acagaaacaa ttcttcttgc cgcaattaca acaagcaagc aagtgagcaa aactgggcta
     1621 attacagtgc agaacaaaat cgaatggggc aggcgggaag caccatctct aactcccatg
     1681 cacagccttt tgatttcccc gatgataacc agaattcaaa aaaactagct gctggacatg
     1741 aattacagcc actagccatt gtggaccagc gaccttcaag cagagccagc agtcgtgcca
     1801 gcagcagacc tcggcctgat gacctggaga tctacccagc tttcttgtac aaagtggttc
     1861 gatgggatcc accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     1921 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1981 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     2041 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     2101 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2161 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     2221 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2281 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     2341 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     2401 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2461 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     2521 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2581 gcatggacga gctgtacaag taaagcggcc gcgactctag atcataatca gccataccac
     2641 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     2701 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     2761 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2821 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa
     2881 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     2941 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     3001 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     3061 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     3121 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     3181 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     3241 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     3301 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3361 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3421 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     3481 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3541 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3601 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3661 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3721 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3781 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     3841 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3901 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3961 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     4021 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4081 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4141 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4201 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4261 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4321 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     4381 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4441 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4501 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4561 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4621 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4681 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4741 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4801 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     4861 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     4921 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     4981 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     5041 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     5101 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5161 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5221 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5281 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5341 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5401 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5461 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5521 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5581 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5641 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5701 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5761 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5821 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
//
