LOCUS Exported 5893 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses rat Cx43 (Gja1 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 Cx43-msfGFP 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5893 /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" regulatory 1833..1842 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1839..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 ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL 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" 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" primer_bind 3667..3686 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" 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 cagtccaagc 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 //