LOCUS       sequence-183151-        6059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Gap Junctions, Excitation: 555, Emission: 584.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6059)
  AUTHORS   Shaner NC, Lin MZ, McKeown MR, Steinbach PA, Hazelwood KL, Davidson 
            MW, Tsien RY
  TITLE     Improving the photostability of bright monomeric orange and red 
            fluorescent proteins.
  JOURNAL   Nat Methods. 2008 Jun;5(6):545-51. doi: 10.1038/nmeth.1209. Epub 
            2008 May 4.
  PUBMED    18454154
REFERENCE   2  (bases 1 to 6059)
  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"; doi: 
            "10.1038/nmeth.1209"; journalName: "Nat Methods"; date: "2008-06"; 
            volume: "5"; issue: "6"; pages: "545-51"
COMMENT     Sequence Label: mTagRFP-T-Cx43-7
FEATURES             Location/Qualifiers
     source          1..6059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1925..1934
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1931..2662
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     polyA_signal    2787..2908
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2824..2843)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2878..2897
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2915..3370)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3052..3073)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3264..3283
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3397..3501
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3503..3860
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3503..3694
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3526..3546)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3711..3846
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3895..4689
                     /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             3895..4689
                     /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(3949..3968)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4559..4578
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4877..4896)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4921..4968
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5297..5885
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5786..5805
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcggcagc acttttcttt cattgggggg
      601 aaggcgtgag gaaagtacca aacagcagca gacttttaaa ctttaaacag acaggtctga
      661 gagcctgaac tctcattttt cctttgactt cagcctccaa ggagttccac caactttggc
      721 gcgccggctt cactttcatt aagtgaaaga gaggtgccca gacatgggtg actggagtgc
      781 cttggggaag cttctggaca aggtccaagc ctactccacc gctggaggga aggtgtggct
      841 gtcagtgctc ttcatattca gaatcctgct cctggggaca gctgttgagt cagcttgggg
      901 tgatgaacag tctgcctttc gctgtaacac tcaacaacct ggctgcgaaa acgtctgcta
      961 tgacaagtcc ttccccatct ctcacgtgcg cttctgggtc cttcagatca tattcgtgtc
     1021 tgtgcccaca ctcctgtact tggcccatgt gttctatgtg atgaggaagg aagagaagct
     1081 aaacaagaaa gaagaggagc tcaaagtggc ccagactgac ggggtcaacg tggagatgca
     1141 cctgaagcag attgaaatca agaagttcaa gtacgggatt gaagagcacg gcaaggtgaa
     1201 aatgaggggc ggcttgctga gaacctacat catcagcatc ctcttcaagt ctgtcttcga
     1261 ggtggccttc ctgctcatcc agtggtacat ctatgggttc agcttgagcg cggtctacac
     1321 ctgcaagaga gatccctgcc cccaccaggt agactgcttc ctctcacgtc ccacggagaa
     1381 aaccatcttc atcatcttca tgctggtggt gtccttggtg tctctcgctt tgaacatcat
     1441 tgagctcttc tacgtcttct tcaagggcgt taaggatcgc gtgaagggaa gaagcgatcc
     1501 ttaccacgcc accactggcc cactgagccc atcaaaagac tgcggatctc caaaatacgc
     1561 ctacttcaat ggctgctcct caccaacggc tccactctcg cctatgtctc ctcctgggta
     1621 caagctggtt actggtgaca gaaacaattc ctcgtgccgc aattacaaca agcaagctag
     1681 cgagcaaaac tgggcgaact acagcgcaga gcaaaatcgc atggggcagg ccggaagcac
     1741 catctccaac tcgcacgccc agccgttcga tttccccgac gacaaccaga atgccaaaaa
     1801 agttgctgct ggacatgaac tccagccatt agccatcgtg gaccaacgac cttccagcag
     1861 agccagcagc cgcgccagca gcaggcctcg gcctgatgac ctggagattg cggatccacc
     1921 ggtcgccacc atggtgtcta agggcgaaga gctgattaag gagaacatgc acatgaagct
     1981 gtacatggag ggcaccgtga acaaccacca cttcaagtgc acatccgagg gcgaaggcaa
     2041 gccctacgag ggcacccaga ccatgagaat caaggtggtc gagggcggcc ctctcccctt
     2101 cgccttcgac atcctggcta ccagcttcat gtacggcagc agaaccttca tcaaccacac
     2161 ccagggcatc cccgatttct ttaagcagtc cttccctgag ggcttcacat gggagagagt
     2221 caccacatac gaagacgggg gcgtgctgac cgctacccag gacaccagcc tccaggacgg
     2281 ctgcctcatc tacaacgtca agatcagagg ggtgaacttc ccatccaacg gccctgtgat
     2341 gcagaagaaa acactcggct gggaggccaa caccgagatg ctgtaccccg ctgacggcgg
     2401 cctggaaggc agaaccgaca tggccctgaa gctcgtgggc gggggccacc tgatctgcaa
     2461 cttcaagacc acatacagat ccaagaaacc cgctaagaac ctcaagatgc ccggcgtcta
     2521 ctatgtggac cacagactgg aaagaatcaa ggaggccgac aaagagacct acgtcgagca
     2581 gcacgaggtg gctgtggcca gatactgcga cctccctagc aaactggggc acaaacttaa
     2641 tggcatggac gagctgtaca agtaagcggc cgcgactcta gatcataatc agccatacca
     2701 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     2761 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     2821 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2881 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     2941 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3001 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3061 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3121 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3181 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3241 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3301 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     3361 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3421 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3481 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     3541 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3601 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3661 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3721 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3781 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3841 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3901 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3961 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4021 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     4081 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4141 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4201 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4261 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4321 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4381 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     4441 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4501 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4561 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4621 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4681 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4741 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4801 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4861 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     4921 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     4981 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     5041 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     5101 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     5161 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5221 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5281 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5341 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5401 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5461 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5521 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5581 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5641 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5701 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5761 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5821 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5881 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5941 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt
     6001 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccg
//
