LOCUS       sequence-128062-        5893 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses rat Cx43 (Gja1 CDS) tagged with non-monomerized sfGFP on 
            the C-terminus. CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5893)
  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 5893)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: Cx43-sfGFP
FEATURES             Location/Qualifiers
     source          1..5893
                     /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      1833..1842
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1839..1841,1842..1844,1845..2558)
                     /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
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1884..1905)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2492..2513
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2681..2802
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2718..2737)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2772..2791
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2809..3264)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2946..2967)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3158..3177
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3291..3395
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3397..3754
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3397..3588
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3420..3440)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3605..3740
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3789..4583
                     /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             3789..4583
                     /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(3843..3862)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4453..4472
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4771..4790)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4815..4862
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5191..5779
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5680..5699
                     /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 ccatcaaaag
     1441 actgcggatc tccaaaatac gcctacttca atggctgctc ctcaccaacg gctccactct
     1501 cgcctatgtc tcctcctggg tacaagctgg ttactggtga cagaaacaat tcctcgtgcc
     1561 gcaattacaa caagcaagct agcgagcaaa actgggcgaa ctacagcgca gagcaaaatc
     1621 gcatggggca ggccggaagc accatctcca actcgcacgc ccagccgttc gatttccccg
     1681 acgacaacca gaatgccaaa aaagttgctg ctggacatga actccagcca ttagccatcg
     1741 tggaccaacg accttccagc agagccagca gccgcgccag cagcaggcct cggcctgatg
     1801 acctggagat ttccgcccgg gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc
     1861 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1921 tcagcgtgcg cggcgagggc gagggcgatg ccaccaacgg caagctgacc ctgaagttca
     1981 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     2041 gcgtgcagtg cttcagccgc taccccgacc acatgaagcg ccacgacttc ttcaagtccg
     2101 ccatgcccga aggctacgtc caggagcgca ccatcagctt caaggacgac ggcacctaca
     2161 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     2221 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aacttcaaca
     2281 gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga
     2341 tccgccacaa cgtggaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     2401 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgtgc
     2461 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     2521 ccgggatcac tcacggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat
     2581 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2641 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2701 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2761 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     2821 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2881 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2941 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3001 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3061 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3121 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3181 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3241 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3301 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3361 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     3421 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3481 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3541 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3601 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3661 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3721 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     3781 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3841 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3901 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3961 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4021 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4081 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4141 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4201 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4261 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     4321 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4381 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4441 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4501 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4561 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4621 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4681 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4741 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     4801 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     4861 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     4921 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     4981 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     5041 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5101 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5161 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5221 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5281 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     5341 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5401 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5461 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5521 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5581 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5641 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5701 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5761 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5821 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5881 taccgccatg cat
//
