LOCUS       sequence-152441-        5993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of C-terminal fusion of mClover3 to CX43.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5993)
  AUTHORS   Bajar BT, Wang ES, Lam AJ, Kim BB, Jacobs CL, Howe ES, Davidson MW, 
            Lin MZ, Chu J
  TITLE     Improving brightness and photostability of green and red fluorescent
            proteins for live cell imaging and FRET reporting.
  JOURNAL   Sci Rep. 2016 Feb 16;6:20889. doi: 10.1038/srep20889.
  PUBMED    26879144
REFERENCE   2  (bases 1 to 5993)
  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: "Sci Rep."; 
            date: "2016-02-16"; volume: "6:20889. doi"; pages: " 
            10.1038/srep20889"
COMMENT     Sequence Label: pKanCMV-CX4.3-7aa-mClover3
FEATURES             Location/Qualifiers
     source          1..5993
                     /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      1985..1994
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1991..1993,1994..1996,1997..2710)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(2036..2057)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2644..2665
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2839..2960
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2876..2895)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2930..2949
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2967..3422)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3104..3125)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3316..3335
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3449..3553
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3555..3912
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3555..3746
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3578..3598)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3763..3898
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3947..4741
                     /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             3947..4741
                     /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(4001..4020)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4611..4630
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4929..4948)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4973..5020
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5349..5937
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5838..5857
                     /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 attcggcagc acttttcttt cattgggggg
      661 aaggcgtgag gaaagtacca aacagcagca gacttttaaa ctttaaacag acaggtctga
      721 gagcctgaac tctcattttt cctttgactt cagcctccaa ggagttccac caactttggc
      781 gcgccggctt cactttcatt aagtgaaaga gaggtgccca gacatgggtg actggagtgc
      841 cttggggaag cttctggaca aggtccaagc ctactccacc gctggaggga aggtgtggct
      901 gtcagtgctc ttcatattca gaatcctgct cctggggaca gctgttgagt cagcttgggg
      961 tgatgaacag tctgcctttc gctgtaacac tcaacaacct ggctgcgaaa acgtctgcta
     1021 tgacaagtcc ttccccatct ctcacgtgcg cttctgggtc cttcagatca tattcgtgtc
     1081 tgtgcccaca ctcctgtact tggcccatgt gttctatgtg atgaggaagg aagagaagct
     1141 aaacaagaaa gaagaggagc tcaaagtggc ccagactgac ggggtcaacg tggagatgca
     1201 cctgaagcag attgaaatca agaagttcaa gtacgggatt gaagagcacg gcaaggtgaa
     1261 aatgaggggc ggcttgctga gaacctacat catcagcatc ctcttcaagt ctgtcttcga
     1321 ggtggccttc ctgctcatcc agtggtacat ctatgggttc agcttgagcg cggtctacac
     1381 ctgcaagaga gatccctgcc cccaccaggt agactgcttc ctctcacgtc ccacggagaa
     1441 aaccatcttc atcatcttca tgctggtggt gtccttggtg tctctcgctt tgaacatcat
     1501 tgagctcttc tacgtcttct tcaaaggcgt taaggatcgc gtgaagggaa gaagcgatcc
     1561 ttaccacgcc accactggcc cactgagccc atcaaaagac tgcggatctc caaaatacgc
     1621 ctacttcaat ggctgctcct caccaacggc tccactctcg cctatgtctc ctcctgggta
     1681 caagctggtt actggtgaca gaaacaattc ctcgtgccgc aattacaaca agcaagctag
     1741 cgagcaaaac tgggcgaact acagcgcaga gcaaaatcgc atggggcagg ccggaagcac
     1801 catctccaac tcgcacgccc agccgttcga tttccccgac gacaaccaga atgccaaaaa
     1861 agttgctgct ggacatgaac tccagccatt agccatcgtg gaccaacgac cttccagcag
     1921 agccagcagc cgcgccagca gcaggcctcg gcctgatgac ctggagattg cggatccacc
     1981 ggtcgccacc atggtgtcta agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     2041 cgagctggac ggcgacgtaa acggccacaa gttcagcgtc cgcggcgagg gcgagggcga
     2101 tgccaccaac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     2161 ctggcccacc ctcgtgacca ccttcggcta cggcgtggcc tgcttcagcc gctaccccga
     2221 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     2281 caccatctct ttcaaggacg acggtaccta caagacccgc gccgaggtga agttcgaggg
     2341 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2401 cctggggcac aagctggagt acaacttcaa cagccactac gtctatatca cggccgacaa
     2461 gcagaagaac tgcatcaagg ctaacttcaa gatccgccac aacgttgagg acggcagcgt
     2521 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2581 cgacaaccac tacctgagcc atcagtccaa gctgagcaaa gaccccaacg agaagcgcga
     2641 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatt acacatggca tggacgagct
     2701 gtacaagtaa gcggccgcga ctctagataa ctgatcataa tcagccatac cacatttgta
     2761 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     2821 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     2881 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     2941 aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     3001 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     3061 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     3121 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     3181 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     3241 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     3301 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     3361 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     3421 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3481 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3541 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     3601 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3661 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3721 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3781 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3841 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3901 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3961 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4021 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4081 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     4141 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4201 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4261 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4321 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4381 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4441 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     4501 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4561 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4621 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4681 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4741 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4801 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4861 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     4921 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     4981 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     5041 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     5101 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     5161 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     5221 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5281 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5341 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5401 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5461 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     5521 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5581 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5641 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5701 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5761 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5821 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5881 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5941 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tca
//
