LOCUS       sequence-124708-        5518 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses rat Cx43 (Gja1 CDS) truncated at amino acid 258 and tagged
            with msfGFP on the C-terminus. CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5518)
  AUTHORS   Stout RF Jr, Snapp EL, Spray DC
  TITLE     Connexin Type and Fluorescent Protein-fusion Tag Determine 
            Structural Stability of Gap Junction Plaques.
  JOURNAL   J Biol Chem. 2015 Aug 11. pii: jbc.M115.659979.
  PUBMED    26265468
REFERENCE   2  (bases 1 to 5518)
  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. 2015 Aug 11. pii: jbc.M115.659979."
COMMENT     Sequence Label: Cx43K258stop-msfGFP
FEATURES             Location/Qualifiers
     source          1..5518
                     /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      1458..1467
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1464..1466,1467..1469,1470..2183)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1509..1530)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2117..2138
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2306..2427
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2343..2362)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2397..2416
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2434..2889)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2571..2592)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2783..2802
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2916..3020
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3022..3379
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3022..3213
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3045..3065)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3230..3365
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3414..4208
                     /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             3414..4208
                     /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(3468..3487)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4078..4097
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4396..4415)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4440..4487
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4816..5404
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5305..5324
                     /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 attctgcagt cgacggtacc gcgggcccgg
      661 gcaccatggg tgactggagt gccttgggga agcttctgga caaggtccaa gcctactcca
      721 ccgctggagg gaaggtgtgg ctgtcagtgc tcttcatatt cagaatcctg ctcctgggga
      781 cagctgttga gtcagcttgg ggtgatgaac agtctgcctt tcgctgtaac actcaacaac
      841 ctggctgcga aaacgtctgc tatgacaagt ccttccccat ctctcacgtg cgcttctggg
      901 tccttcagat catattcgtg tctgtgccca ccctcctgta cttggcccat gtgttctatg
      961 tgatgaggaa ggaagagaag ctaaacaaga aagaagagga gctcaaagtg gcccagactg
     1021 acggggtcaa cgtggagatg cacctgaagc agattgaaat caagaagttc aagtacggga
     1081 ttgaagagca cggcaaggtg aaaatgaggg gcggcttgct gagaacctac atcatcagca
     1141 tcctcttcaa gtctgtcttc gaggtggcct tcctgctcat ccagtggtac atctatgggt
     1201 tcagcttgag cgcggtctac acctgcaaga gagatccctg cccccaccag gtagactgct
     1261 tcctctcacg tcccacggag aaaaccatct tcatcatctt catgctggtg gtgtccttgg
     1321 tgtctctcgc tttgaacatc attgagctct tctacgtctt cttcaaaggc gttaaggatc
     1381 gcgtgaaggg aagaagcgat ccttaccacg ccaccactgg cccactgagc ccatcatccg
     1441 cccgggatcc accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     1501 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgcgcggcg
     1561 agggcgaggg cgatgccacc aacggcaagc tgaccctgaa gttcatctgc accaccggca
     1621 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     1681 gccgctaccc cgaccacatg aagcgccacg acttcttcaa gtccgccatg cccgaaggct
     1741 acgtccagga gcgcaccatc agcttcaagg acgacggcac ctacaagacc cgcgccgagg
     1801 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1861 aggacggcaa catcctgggg cacaagctgg agtacaactt caacagccac aacgtctata
     1921 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacgtgg
     1981 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2041 ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca
     2101 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactcacg
     2161 gcatggacga gctgtacaag taaagcggcc gcgactctag atcataatca gccataccac
     2221 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     2281 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     2341 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2401 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa
     2461 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     2521 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     2581 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     2641 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     2701 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     2761 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     2821 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     2881 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2941 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3001 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     3061 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3121 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3181 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3241 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3301 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3361 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     3421 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3481 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3541 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     3601 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3661 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3721 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     3781 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     3841 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     3901 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     3961 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4021 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4081 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4141 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4201 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4261 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4321 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4381 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     4441 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     4501 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     4561 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     4621 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     4681 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4741 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4801 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4861 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4921 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     4981 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5041 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5101 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5161 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5221 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5281 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5341 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5401 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5461 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
