LOCUS       sequence-233378-        5465 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For fusion of TagRFP657 to N-term of insert. SEE DEPOSITOR COMMENTS 
            BELOW..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5465)
  AUTHORS   Morozova KS, Piatkevich KD, Gould TJ, Zhang J, Bewersdorf J, 
            Verkhusha VV
  TITLE     Far-red fluorescent protein excitable with red lasers for flow 
            cytometry and superresolution STED nanoscopy.
  JOURNAL   Biophys J. 2010 Jul 21;99(2):L13-5.
  PUBMED    20643047
REFERENCE   2  (bases 1 to 5465)
  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: "Biophys 
            J."; date: "2010-07-21"; volume: "99(2)"; pages: "L13-5"
COMMENT     Sequence Label: pTagRFP657-C1
FEATURES             Location/Qualifiers
     source          1..5465
                     /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      601..610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             607..1317
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKDNMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTQ
                     RIKVVEGGPLPFAFDILATSFMYGSHTFINHTQGIPDFWKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEAHTEMLYPADGGLEGRTA
                     LALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDYRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     regulatory      1347..1356
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1371..2063
                     /codon_start=1
                     /product="far-red monomeric fluorescent protein with an 
                     emission peak at 657 nm (Morozova et al., 2010)"
                     /label=TagRFP657
                     /note="derived from mKate"
                     /translation="ELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTQRIKVVE
                     GGPLPFAFDILATSFMYGSHTFINHTQGIPDFWKQSFPEGFTWERVTTYEDGGVLTATQ
                     DTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEAHTEMLYPADGGLEGRTALALKLV
                     GGGHLICNFKTTYRSKKPAKNLKMPGVYYVDYRLERIKEADKETYVEQHEVAVARYCDL
                     PSKLGHKLN"
     misc_feature    2064..2129
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2253..2374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2290..2309)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2344..2363
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2381..2836)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2518..2539)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2730..2749
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2863..2967
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2969..3326
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2969..3160
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2992..3012)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3177..3312
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3361..4155
                     /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             3361..4155
                     /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(3415..3434)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4025..4044
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4343..4362)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4387..4434
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4763..5351
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5252..5271
                     /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 gctagcggtg
      601 gccaccatgg tgtctaaggg cgaagagctg attaaggaca acatgcacat gaagctgtac
      661 atggagggca ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga aggcaagccc
      721 tacgagggca cccagaccca gagaatcaag gtggtcgagg gcggccctct ccccttcgcc
      781 ttcgacatcc tggctaccag cttcatgtac ggcagtcaca ccttcatcaa ccacacccag
      841 ggcatccccg acttctggaa gcagtccttc cctgagggct tcacatggga gagagtcacc
      901 acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggctgc
      961 ctcatctaca acgtcaagat cagaggggtg aacttcccat ccaacggccc tgtgatgcag
     1021 aagaaaacac tcggctggga ggcccacacc gagatgctgt accccgctga cggcggcctg
     1081 gaaggcagaa ccgcgctggc cctgaagctc gtgggcgggg gccacctgat ctgcaacttc
     1141 aagaccacat acagatccaa gaaacccgct aagaacctca agatgcccgg cgtctactat
     1201 gtggactaca gactggaaag aatcaaggag gccgacaaag agacctacgt cgagcagcac
     1261 gaggtggctg tggccagata ctgcgacctc cctagcaaac tggggcacaa gcttaattcc
     1321 ggactcagat ctcatagcta gcggtggcca ccatggtgtc taagagcgaa gagctgatta
     1381 aggagaacat gcacatgaag ctgtacatgg agggcaccgt gaacaaccac cacttcaagt
     1441 gcacatccga gggcgaaggc aagccctacg agggcaccca gacccagaga atcaaggtgg
     1501 tcgagggcgg ccctctcccc ttcgccttcg acatcctggc taccagcttc atgtacggca
     1561 gtcacacctt catcaaccac acccagggca tccccgactt ctggaagcag tccttccctg
     1621 agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg accgctaccc
     1681 aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga ggggtgaact
     1741 tcccatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc cacaccgaga
     1801 tgctgtaccc cgctgacggc ggcctggaag gcagaaccgc gctggccctg aagctcgtgg
     1861 gcgggggcca cctgatctgc aacttcaaga ccacatacag atccaagaaa cccgctaaga
     1921 acctcaagat gcccggcgtc tactatgtgg actacagact ggaaagaatc aaggaggccg
     1981 acaaagagac ctacgtcgag cagcacgagg tggctgtggc cagatactgc gacctcccta
     2041 gcaaactggg gcacaagctt aattccggac tcagatctcg agctcaagct tcgaattctg
     2101 cagtcgacgg taccgcgggc ccgggatcca ccggatctag ataactgatc ataatcagcc
     2161 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     2221 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     2281 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2341 gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt
     2401 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     2461 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     2521 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     2581 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     2641 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     2701 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     2761 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     2821 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     2881 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2941 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     3001 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3061 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3121 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3181 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3241 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3301 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     3361 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3421 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3481 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3541 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3601 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3661 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3721 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     3781 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     3841 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     3901 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     3961 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4021 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4081 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4141 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4201 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4261 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4321 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     4381 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     4441 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     4501 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     4561 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     4621 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4681 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4741 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4801 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4861 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4921 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4981 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5041 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5101 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5161 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5221 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5281 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5341 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5401 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     5461 tgcat
//
