LOCUS       sequence-127981-        5187 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Cx30 (GJB6 CDS) with a 7 amino acid linker on the 
            C-terminus of the Connexin linking to monomerized superfolder Green 
            Fluorescent Protein. Expression driven by CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5187)
  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 5187)
  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: Cx30-msfGFP
FEATURES             Location/Qualifiers
     source          1..5187
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        246..297
                     /label=CMVd2 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields moderately reduced expression 
                     relative to the full-length promoter"
     primer_bind     248..268
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1127..1136
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1133..1135,1136..1138,1139..1852)
                     /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(1178..1199)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1786..1807
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1975..2096
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2012..2031)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2066..2085
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2103..2558)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2240..2261)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2452..2471
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2585..2689
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2691..3048
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2691..2882
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2714..2734)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2899..3034
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3083..3877
                     /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             3083..3877
                     /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(3137..3156)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3747..3766
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4065..4084)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4109..4156
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4485..5073
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4974..4993
                     /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 atgggagttt gtttggcacc aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      241 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
      301 ttagtgaacc gtcagatccg ctagcatgga ttgggggacg ctgcacactt tcatcggggg
      361 tgtcaacaaa cactccacca gcatcgggaa ggtgtggatc acagtcatct ttattttccg
      421 agtcatgatc ctcgtggtgg ctgcccagga agtgtggggt gacgagcaag aggacttcgt
      481 ctgcaacaca ctgcaaccgg gatgcaaaaa tgtgtgctat gaccactttt tcccggtgtc
      541 ccacatccgg ctgtgggccc tccagctgat cttcgtctcc accccagcgc tgctggtggc
      601 catgcatgtg gcctactaca ggcacgaaac cactcgcaag ttcaggcgag gagagaagag
      661 gaatgatttc aaagacatag aggacattaa aaagcagaag gttcggatag aggggtcgct
      721 gtggtggacg tacaccagca gcatcttttt ccgaatcatc tttgaagcag cctttatgta
      781 tgtgttttac ttcctttaca atgggtacca cctgccctgg gtgttgaaat gtgggattga
      841 cccctgcccc aaccttgttg actgctttat ttctaggcca acagagaaga ccgtgtttac
      901 catttttatg atttctgcgt ctgtgatttg catgctgctt aacgtggcag agttgtgcta
      961 cctgctgctg aaagtgtgtt ttaggagatc aaagagagca cagacgcaaa aaaatcaccc
     1021 caatcatgcc ctaaaggaga gtaagcagaa tgaaatgaat gagctgattt cagatagtgg
     1081 tcaaaatgca atcacaggtt tcccaagcta cccggatcca ccggtcgcca ccatggtgag
     1141 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1201 aaacggccac aagttcagcg tgcgcggcga gggcgagggc gatgccacca acggcaagct
     1261 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1321 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcgccacga
     1381 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatca gcttcaagga
     1441 cgacggcacc tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1501 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1561 gtacaacttc aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa
     1621 ggccaacttc aagatccgcc acaacgtgga ggacggcagc gtgcagctcg ccgaccacta
     1681 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1741 cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1801 gttcgtgacc gccgccggga tcactcacgg catggacgag ctgtacaagt aaagcggccg
     1861 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     1921 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     1981 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2041 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     2101 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     2161 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     2221 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     2281 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     2341 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     2401 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     2461 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     2521 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     2581 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2641 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     2701 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2761 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2821 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2881 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2941 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3001 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     3061 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3121 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3181 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3241 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     3301 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3361 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3421 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3481 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3541 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3601 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     3661 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     3721 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     3781 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     3841 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     3901 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     3961 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4021 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     4081 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     4141 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     4201 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     4261 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     4321 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4381 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4441 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4501 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4561 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4621 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4681 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4741 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4801 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4861 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4921 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4981 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5041 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5101 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5161 gtggataacc gtattaccgc catgcat
//
