LOCUS       sequence-388910-        5675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing fluorescent SNAPf-mTurquoise2 fusion protein in mammalian
            cells with localization in the nucleus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5675)
  AUTHORS   Martin A, Rivera-Fuentes P
  TITLE     A general strategy to develop fluorogenic polymethine dyes for 
            bioimaging.
  JOURNAL   Nat Chem. 2024 Jan;16(1):28-35. doi: 10.1038/s41557-023-01367-y. 
            Epub 2023 Nov 27.
  PUBMED    38012391
REFERENCE   2  (bases 1 to 5675)
  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/s41557-023-01367-y"; journalName: "Nat Chem"; date: 
            "2024-01"; volume: "16"; issue: "1"; pages: "28-35"
COMMENT     Sequence Label: H2B-SNAPf-mTurquoise2
FEATURES             Location/Qualifiers
     source          1..5675
                     /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"
     CDS             610..987
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             1042..1587
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     CDS             join(1624..1626,1627..2340)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1666..1687)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1927..1946)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2274..2295
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2463..2584
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2500..2519)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2554..2573
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2591..3046)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2728..2749)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2940..2959
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3073..3177
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3179..3536
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3179..3370
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3202..3222)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3387..3522
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3571..4365
                     /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             3571..4365
                     /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(3625..3644)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4235..4254
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4553..4572)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4597..4644
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4973..5561
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5462..5481
                     /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 ccggactcaa tgccagagcc agcgaagtct gctcccgccc cgaaaaaggg ctccaagaag
      661 gcggtgacta aggcgcagaa gaaaggcggc aagaagcgca agcgcagccg caaggagagc
      721 tattccatct atgtgtacaa ggttctgaag caggtccacc ctgacaccgg catttcgtcc
      781 aaggccatgg gcatcatgaa ttcgtttgtg aacgacattt tcgagcgcat cgcaggtgag
      841 gcttcccgcc tggcgcatta caacaagcgc tcgaccatca cctccaggga gatccagacg
      901 gccgtgcgcc tgctgctgcc tggggagttg gccaagcacg ccgtgtccga gggtactaag
      961 gccatcacca agtacaccag cgctaaggct agcgatatcg gcgcgccagc atttaaatct
     1021 gtacagaccg gtgaattcac catggacaaa gactgcgaaa tgaagcgcac caccctggat
     1081 agccctctgg gcaagctgga actgtctggg tgcgaacagg gcctgcaccg tatcatcttc
     1141 ctgggcaaag gaacatctgc cgccgacgcc gtggaagtgc ctgccccagc cgccgtgctg
     1201 ggcggaccag agccactgat gcaggccacc gcctggctca acgcctactt tcaccagcct
     1261 gaggccatcg aggagttccc tgtgccagcc ctgcaccacc cagtgttcca gcaggagagc
     1321 tttacccgcc aggtgctgtg gaaactgctg aaagtggtga agttcggaga ggtcatcagc
     1381 tacagccacc tggccgccct ggccggcaat cccgccgcca ccgccgccgt gaaaaccgcc
     1441 ctgagcggaa atcccgtgcc cattctgatc ccctgccacc gggtggtgca gggcgacctg
     1501 gacgtggggg gctacgaggg cgggctcgcc gtgaaagagt ggctgctggc ccacgagggc
     1561 cacagactgg gcaagcctgg gctgggtcct gcaggcggat ccgcgtttaa actcgaggtt
     1621 aatgtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1681 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1741 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1801 ctcgtgacca ccctgtcctg gggcgtgcag tgcttcgccc gctaccccga ccacatgaag
     1861 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     1921 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1981 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     2041 aagctggagt acaactactt tagcgacaac gtctatatca ccgccgacaa gcagaagaac
     2101 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
     2161 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     2221 tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     2281 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     2341 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     2401 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     2461 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     2521 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     2581 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     2641 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     2701 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     2761 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     2821 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     2881 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     2941 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     3001 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     3061 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3121 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     3181 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3241 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3301 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3361 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3421 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     3481 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     3541 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3601 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3661 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3721 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     3781 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3841 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3901 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3961 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4021 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4081 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4141 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4201 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4261 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4321 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4381 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4441 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4501 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     4561 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     4621 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     4681 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     4741 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     4801 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     4861 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4921 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4981 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5041 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5101 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5161 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5221 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5281 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5341 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5401 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5461 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5521 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5581 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5641 ctgattctgt ggataaccgt attaccgcca tgcat
//
