LOCUS       sequence-221558-        5635 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 5635)
  TITLE     Davidson Orange Fluorescent Proteins
REFERENCE   2  (bases 1 to 5635)
  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"
COMMENT     Sequence Label: mTagRFP-T-Cx32-7
FEATURES             Location/Qualifiers
     source          1..5635
                     /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      1501..1510
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1507..2238
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     polyA_signal    2363..2484
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2400..2419)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2454..2473
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2491..2946)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2628..2649)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2840..2859
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2973..3077
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3079..3436
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3079..3270
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3102..3122)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3287..3422
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3471..4265
                     /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             3471..4265
                     /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(3525..3544)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4135..4154
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4453..4472)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4497..4544
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4873..5461
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5362..5381
                     /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 attccctctg ggaaagggca gcagcagcca
      601 ggtgtggcag tgacagggag gtgtgaatga ggcaggatga actggacagg tttgtacacc
      661 ttgctcagtg gcgtgaaccg gcattctact gccattggcc gagtatggct ctcggtcatc
      721 ttcatcttca gaatcatggt gctggtggtg gctgcagaga gtgtgtgggg tgatgagaaa
      781 tcttccttca tctgcaacac actccagcct ggctgcaaca gcgtttgcta tgaccaattc
      841 ttccccatct cccatgtgcg gctgtggtcc ctgcagctca tcctagtttc caccccagct
      901 ctcctcgtgg ccatgcacgt ggctcaccag caacacatag agaagaaaat gctacggctt
      961 gagggccatg gggaccccct acacctggag gaggtgaaga ggcacaaggt ccacatctca
     1021 gggacactgt ggtggaccta tgtcatcagc gtggtgttcc ggctgttgtt tgaggccgtc
     1081 ttcatgtatg tcttttatct gctctaccct ggctatgcca tggtgcggct ggtcaagtgc
     1141 gacgtctacc cctgccccaa cacagtggac tgcttcgtgt cccgccccac cgagaaaacc
     1201 gtcttcaccg tcttcatgct agctgcctct ggcatctgca tcatcctcaa tgtggccgag
     1261 gtggtgtacc tcatcatccg ggcctgtgcc cgccgagccc agcgccgctc caatccacct
     1321 tcccgcaagg gctcgggctt cggccaccgc ctctcacctg aatacaagca gaatgagatc
     1381 aacaagctgc tgagtgagca ggatggctcc ctgaaagaca tactgcgccg cagccctggc
     1441 accggggctg ggctggctga aaagagcgac cgctgctcgg cctgcgcgga tccaccggtc
     1501 gccaccatgg tgtctaaggg cgaagagctg attaaggaga acatgcacat gaagctgtac
     1561 atggagggca ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga aggcaagccc
     1621 tacgagggca cccagaccat gagaatcaag gtggtcgagg gcggccctct ccccttcgcc
     1681 ttcgacatcc tggctaccag cttcatgtac ggcagcagaa ccttcatcaa ccacacccag
     1741 ggcatccccg atttctttaa gcagtccttc cctgagggct tcacatggga gagagtcacc
     1801 acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggctgc
     1861 ctcatctaca acgtcaagat cagaggggtg aacttcccat ccaacggccc tgtgatgcag
     1921 aagaaaacac tcggctggga ggccaacacc gagatgctgt accccgctga cggcggcctg
     1981 gaaggcagaa ccgacatggc cctgaagctc gtgggcgggg gccacctgat ctgcaacttc
     2041 aagaccacat acagatccaa gaaacccgct aagaacctca agatgcccgg cgtctactat
     2101 gtggaccaca gactggaaag aatcaaggag gccgacaaag agacctacgt cgagcagcac
     2161 gaggtggctg tggccagata ctgcgacctc cctagcaaac tggggcacaa acttaatggc
     2221 atggacgagc tgtacaagta agcggccgcg actctagatc ataatcagcc ataccacatt
     2281 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2341 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     2401 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2461 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     2521 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     2581 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     2641 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     2701 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     2761 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     2821 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     2881 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     2941 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3001 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3061 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     3121 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3181 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3241 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3301 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3361 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3421 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3481 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3541 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3601 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     3661 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3721 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3781 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3841 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3901 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3961 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     4021 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4081 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4141 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4201 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4261 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4321 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4381 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     4441 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     4501 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     4561 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     4621 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     4681 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     4741 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     4801 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4861 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4921 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4981 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5041 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5101 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5161 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5221 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5281 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5341 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5401 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5461 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5521 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt
     5581 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccg
//
