LOCUS       sequence-408399-        7524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human Cx43 with S368A mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7524)
  AUTHORS   Horns F, Martinez JA, Fan C, Haque M, Linton JM, Tobin V, Santat L, 
            Maggiolo AO, Bjorkman PJ, Lois C, Elowitz MB
  TITLE     Engineering RNA export for measurement and manipulation of living 
            cells.
  JOURNAL   Cell. 2023 Jul 10:S0092-8674(23)00689-X. doi: 
            10.1016/j.cell.2023.06.013.
  PUBMED    37437570
REFERENCE   2  (bases 1 to 7524)
  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: "Cell. 2023 
            Jul 10:S0092-8674(23)00689-X. doi: 10.1016/j.cell.2023.06.013."
COMMENT     Sequence Label: pJAM1.16_PB-Cx43_S368A-PGK-Blast
FEATURES             Location/Qualifiers
     source          1..7524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   14..48
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        660..871
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             884..1152
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      1180..1189
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2462..2583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2499..2518)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2553..2572
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2956..2975
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      3053..3062
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3059..3457
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3475..3699
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4126..4188
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4687..4703)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4687..4703)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4700..4722)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4711..4727
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4735..4741,4742..4759,4760..4765))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4780..4801
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4918..4935)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5089..5677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5169..5188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5848..6642)
                     /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             complement(5848..6642)
                     /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(5959..5978)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6569..6588
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6643..6747)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6773..7228)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6910..6931)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7122..7141
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7355..7377
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7369..7386
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7370..7386
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gcagctagat taaccctaga aagatagtct gcgtaaaatt gacgcatgca ttcttgaaat
       61 attgctctct ctttctaaat agcgcgaatc cgtcgctgtg catttaggac atctcagtcg
      121 ccgcttggag ctcccgtgag gcgtgcttgt caatgcggta agtgtcactg attttgaact
      181 ataacaaccg cgtgagtcaa aatgacgcat gattatcttt tacgtgactt ttaagattta
      241 actcatacga taattatatt gttatttcgt gttctactta cgtgataact tattatatat
      301 atattttctt gttatagata tccttgaggg acagcccccc cccaaagccc ccagggatgt
      361 aattacgtcc ctcccccgct agggggcagc agcgagccgc ccggggctcc gctccggtcc
      421 ggcgctcccc ccgcatcccc gagccggcag cgtgcgggga cagcccgggc acggggaagg
      481 tggcacggga tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg cagacacctg
      541 gggggatacg gggaaaaatc gattaactat aacggtccta aggtagcgaa cacctgcgct
      601 cgagcgacgt cattaccctg ttatccctag gatctgcgat cgctccggtg cccgtcagtg
      661 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
      721 gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
      781 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
      841 ttttcgcaac gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc
      901 ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc
      961 gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag
     1021 gtcgagaccg ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc
     1081 cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg
     1141 ccgttacaga tccaagctgt gaccggcgcc tacgctagcg ccaccatggg tgactggagc
     1201 gccttaggca aactccttga caaggttcaa gcctactcaa ctgctggagg gaaggtgtgg
     1261 ctgtcagtac ttttcatttt ccgaatcctg ctgctgggga cagcggttga gtcagcctgg
     1321 ggagatgagc agtctgcctt tcgttgtaac actcagcaac ctggttgtga aaatgtctgc
     1381 tatgacaagt ctttcccaat ctctcatgtg cgcttctggg tcctgcagat catatttgtg
     1441 tctgtaccca cactcttgta cctggctcat gtgttctatg tgatgcgaaa ggaagagaaa
     1501 ctgaacaaga aagaggaaga actcaaggtt gcccaaactg atggtgtcaa tgtggacatg
     1561 cacttgaagc agattgagat aaagaagttc aagtacggta ttgaagagca tggtaaggtg
     1621 aaaatgcgag gggggttgct gcgaacctac atcatcagta tcctcttcaa gtctatcttt
     1681 gaggtggcct tcttgctgat ccagtggtac atctatggat tcagcttgag tgctgtttac
     1741 acttgcaaaa gagatccctg cccacatcag gtggactgtt tcctctctcg ccccacggag
     1801 aaaaccatct tcatcatctt catgctggtg gtgtccttgg tgtccctggc cttgaatatc
     1861 attgaactct tctatgtttt cttcaagggc gttaaggatc gggttaaggg aaagagcgac
     1921 ccttaccatg cgaccagtgg tgcgctgagc cctgccaaag actgtgggtc tcaaaaatat
     1981 gcttatttca atggctgctc ctcaccaacc gctcccctct cgcctatgtc tcctcctggg
     2041 tacaagctgg ttactggcga cagaaacaat tcttcttgcc gcaattacaa caagcaagca
     2101 agtgagcaaa actgggctaa ttacagtgca gaacaaaatc gaatggggca ggcgggaagc
     2161 accatctcta actcccatgc acagcctttt gatttccccg atgataacca gaattcaaaa
     2221 aaactagctg ctggacatga attacagcca ctagccattg tggaccagcg accttcaagc
     2281 agagccgcca gtcgtgccag cagcagacct cggcctgatg acctggagat ctaggcggcc
     2341 gcgtcgacga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2401 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2461 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2521 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2581 ttaagcgccg gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta
     2641 ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag
     2701 tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag
     2761 cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc
     2821 tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa
     2881 ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt
     2941 cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct
     3001 cttcctcatc tccgggcctt tcgacctgca gcccaagctt accatgggta ccgccaccat
     3061 ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa
     3121 cagcatcccc atctctgaag attacagcgt cgccagcgca gctctctcta gcgacggccg
     3181 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt
     3241 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa
     3301 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca
     3361 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg
     3421 tgaattgctg ccctctggtt atgtgtggga gggctaaggt cacccgcgcg cggactgtgc
     3481 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     3541 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     3601 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     3661 acaatagcag gcatgctggg gatgcggtgg gctctatggt tcgaaccagt acgatggttt
     3721 tccccgtatc cccccaggtg tctgcaggct caaagagcag cgagaagcgt tcagaggaaa
     3781 gcgatcccgt gccaccttcc ccgtgcccgg gctgtccccg cacgctgccg gctcggggat
     3841 gcggggggag cgccggaccg gagcggagcc ccgggcggct cgctgctgcc ccctagcggg
     3901 ggagggacgt aattacatcc ctgggggctt tggggggggg ctgtccctga taaaagtttt
     3961 gttactttat agaagaaatt ttgagttttt gttttttttt aataaataaa taaacataaa
     4021 taaattgttt gttgaattta ttattagtat gtaagtgtaa atataataaa acttaatatc
     4081 tattcaaatt aataaataaa cctcgatata cagaccgata aaacacatgc gtcaatttta
     4141 cgcatgatta tctttaacgt acgtcacaat atgattatct ttctagggtt aatctagctg
     4201 cggcgcgcca tcaggatcat atcgtcgggt cttttttccg gctcagtcat cgcccaagct
     4261 ggcgctatct gggcatcggg gaggaagaag cccgtgcctt ttcccgcgag gttgaagcgg
     4321 catggaaaga gtttgccgag gatgactgct gctgcattga cgttgagcga aaacgcacgt
     4381 ttaccatgat gattcgggaa ggtgtggcca tgcacgcctt taacggtgaa ctgttcgttc
     4441 aggccacctg ggataccagt tcgtcgcggc ttttccggac acagttccgg atggtcagcc
     4501 cgaagcgcat cagcaacccg aacaataccg gcgacagccg gaactgccgt gccggtgtgc
     4561 agattaatga cagcggtgcg gcgctgggat attacgtcag cgaggacggg tatcctggct
     4621 ggatgccgca gaaatggaca tggatacccc gtgagttacc cggcgggcgc gcttggcgta
     4681 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4741 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4801 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4861 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4921 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4981 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5041 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5101 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5161 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5221 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5281 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5341 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5401 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5461 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5521 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5581 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5641 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5701 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5761 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5821 tatatatgag taaacttggt ctgacagtca gaagaactcg tcaagaaggc gatagaaggc
     5881 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc
     5941 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc
     6001 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt
     6061 cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt
     6121 gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg
     6181 atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg
     6241 gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat
     6301 ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc
     6361 caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac
     6421 gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc
     6481 ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc
     6541 ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca
     6601 agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atactcttcc tttttcaata
     6661 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6721 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta
     6781 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     6841 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     6901 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     6961 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     7021 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     7081 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     7141 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     7201 gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg
     7261 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     7321 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     7381 gccagtgagc gcgcctcgtt cattcacgtt tttgaacccg tggaggacgg gcagactcgc
     7441 ggtgcaaatg tgttttacag cgtgatggag cagatgaaga tgctcgacac gctgcagaac
     7501 acgagctcca ccgcggggcg cgcc
//
