LOCUS       Exported                5230 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Encodes C-terminal fragment of human connexin 43 initiating at M213 
            with a V5 C-terminal fusion tag.
ACCESSION   .
VERSION     .
KEYWORDS    pDEST-mCherry-GJA1-20k-N1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5230)
  AUTHORS   Smyth JW, Shaw RM
  TITLE     Autoregulation of Connexin43 Gap Junction Formation by Internally 
            Translated Isoforms.
  JOURNAL   Cell Rep. 2013 Nov 5. pii: S2211-1247(13)00578-0. doi: 
            10.1016/j.celrep.2013.10.009.
  PUBMED    24210816
REFERENCE   2  (bases 1 to 5230)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5230
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     protein_bind    602..626
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1143..1167)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1181..1190
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1187..1897
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1332..1350)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1600..1619
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2018..2139
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2055..2074)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2109..2128
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2146..2601)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2283..2304)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2495..2514
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2628..2732
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2734..3091
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2757..2777)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2942..3077
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3004..3023
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3126..3920
                     /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(3180..3199)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3790..3809
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4108..4127)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4152..4199
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4528..5116
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5017..5036
                     /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 gctagcatca
      601 aacaagtttg tacaaaaaag caggcttcac catgctggtg gtgtccttgg tgtccctggc
      661 cttgaatatc attgaactct tctatgtttt cttcaagggc gttaaggatc gggttaaggg
      721 aaagagcgac ccttaccatg cgaccagtgg tgcgctgagc cctgccaaag actgtgggtc
      781 tcaaaaatat gcttatttca atggctgctc ctcaccaacc gctcccctct cgcctctatc
      841 tcctcctggg tacaagctgg ttactggcga cagaaacaat tcttcttgcc gcaattacaa
      901 caagcaagca agtgagcaaa actgggctaa ttacagtgca gaacaaaatc gactagggca
      961 ggcgggaagc accatctcta actcccatgc acagcctttt gatttccccg atgataacca
     1021 gaattcaaaa aaactagctg ctggacatga attacagcca ctagccattg tggaccagcg
     1081 accttcaagc agagccagca gtcgtgccag cagcagacct cggcctgatg acctggagat
     1141 ctacccagct ttcttgtaca aagtggttcg atgggatccc gccaccatgg tgagcaaggg
     1201 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     1261 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     1321 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
     1381 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     1441 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     1501 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     1561 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     1621 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     1681 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     1741 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
     1801 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
     1861 ccactccacc ggcggcatgg acgagctgta caagtagcgg ccgcgactct agatcataat
     1921 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     1981 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     2041 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2101 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     2161 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     2221 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     2281 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     2341 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     2401 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     2461 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     2521 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     2581 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2641 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2701 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     2761 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2821 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2881 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2941 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3001 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3061 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     3121 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3181 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3241 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3301 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3361 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3421 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3481 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3541 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     3601 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     3661 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     3721 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     3781 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     3841 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     3901 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     3961 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4021 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4081 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     4141 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     4201 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     4261 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     4321 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     4381 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4441 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4501 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4561 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4621 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     4681 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4741 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4801 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4861 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4921 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4981 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5041 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5101 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5161 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5221 cgccatgcat
//
