LOCUS       sequence-271529-        5319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Histone 2B labeling with near-infrared fluorescent protein emiRFP703
            (enhanced miRFP703).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5319)
  AUTHORS   Matlashov ME, Shcherbakova DM, Alvelid J, Baloban M, Pennacchietti 
            F, Shemetov AA, Testa I, Verkhusha VV
  TITLE     A set of monomeric near-infrared fluorescent proteins for multicolor
            imaging across scales.
  JOURNAL   Nat Commun. 2020 Jan 13;11(1):239. doi: 10.1038/s41467-019-13897-6.
  PUBMED    31932632
REFERENCE   2  (bases 1 to 5319)
  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: "Nat 
            Commun."; date: "2020-01-13"; volume: "11(1):239. doi"; pages: " 
            10.1038/s41467-019-13897-6"
COMMENT     Sequence Label: pH2B-emiRFP703
FEATURES             Location/Qualifiers
     source          1..5319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        97..400
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        401..604
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     555..575
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             693..1070
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      1083..1092
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2143..2264
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2180..2199)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2234..2253
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2271..2726)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2408..2429)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2620..2639
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2753..2857
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2859..3216
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2859..3050
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2882..2902)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3067..3202
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3251..4045
                     /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             3251..4045
                     /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(3305..3324)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3915..3934
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4233..4252)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4277..4324
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4653..5241
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5142..5161
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
       61 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      121 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      181 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      241 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      301 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      361 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      421 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      481 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      541 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      601 agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatct cgagctcaag
      661 cttcgaattc tgcagtcgac ggtaccgcca ccatgccaga gccagcgaag tctgctcccg
      721 ccccgaaaaa gggctccaag aaggcggtga ctaaggcgca gaagaaaggc ggcaagaagc
      781 gcaagcgcag ccgcaaggag agctattcca tctatgtgta caaggttctg aagcaggtcc
      841 accctgacac cggcatttcg tccaaggcca tgggcatcat gaattcgttt gtgaacgaca
      901 ttttcgagcg catcgcaggt gaggcttccc gcctggcgca ttacaacaag cgctcgacca
      961 tcacctccag ggagatccag acggccgtgc gcctgctgct gcctggggag ttggccaagc
     1021 acgccgtgtc cgagggtact aaggccatca ccaagtacac cagcgctaag gatccaccgg
     1081 tcgccaccat ggcggaaggc tccgtcgcca ggcagcctga cctcttgacc tgcgaacatg
     1141 aagagatcca cctcgccggc tcgatccagc cgcatggcgc gcttctggtc gtcagcgaac
     1201 atgatcatcg cgtcatccag gccagcgcca acgccgcgga atttctgaat ctcggaagcg
     1261 tactcggcgt tccgctcgcc gagatcgacg gcgatctgtt gatcaagatc ctgccgcatc
     1321 tcgatcccac cgccgaaggc atgccggtcg cggtgcgctg ccggatcggc aatccctcta
     1381 cggagtactg cggtctgatg catcggcctc cggaaggcgg gctgatcatc gaactcgaac
     1441 gtgccggccc gtcgatcgat ctgtcaggca cgctggcgcc ggcgctggag cggatccgca
     1501 cggcgggttc actgcgcgcg ctgtgcgatg acaccgtgct gctgtttcag cagtgcaccg
     1561 gctacgaccg ggtgatggtg tatcgtttcg atgagcaagg ccacggcctg gtattctccg
     1621 agtgccatgt gcctgggctc gaatcctatt tcggcaaccg ctatccgtcg tcgctggtcc
     1681 cgcagatggc gcggcagctg tacgtgcggc agcgcgtccg cgtgctggtc gacgtcacct
     1741 atcagccggt gccgctggag ccgcggctgt cgccgctgac cgggcgcgat ctcgacatgt
     1801 cgggctgctt cctgcgctcg atgtcgccga tccatctgca gttcctgaag gacatgggcg
     1861 tgcgcgccac cctggcggtg tcgctggtgg tcggcggcaa gctgtggggc ctggttgtct
     1921 gtcaccatta tctgccgcgc ttcatccgtt tcgagctgcg ggcgatctgc aaacggctcg
     1981 ccgaaaggat cgcgacgcgg atcaccgcgc ttgagagcta agcggccgcg actctagatc
     2041 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     2101 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     2161 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     2221 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag
     2281 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     2341 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     2401 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     2461 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     2521 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     2581 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     2641 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     2701 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     2761 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2821 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     2881 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2941 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3001 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3061 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3121 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3181 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     3241 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3301 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3361 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3421 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3481 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     3541 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     3601 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     3661 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     3721 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     3781 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     3841 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     3901 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     3961 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4021 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4081 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4141 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4201 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     4261 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     4321 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     4381 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     4441 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     4501 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4561 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4621 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4681 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4741 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4801 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4861 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4921 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4981 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5041 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5101 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5161 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5221 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5281 tgctggcctt ttgctcacat gttctttcct gcgttatcc
//
