LOCUS       sequence-410748-        4852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Histone 2B labelled with near-infrared fluorescent protein 
            miRFP704nano.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4852)
  AUTHORS   Oliinyk OS, Pletnev S, Baloban M, Verkhusha VV
  TITLE     Development of bright red-shifted miRFP704nano using structural 
            analysis of miRFPnano proteins.
  JOURNAL   Protein Sci. 2023 Jun 22:e4709. doi: 10.1002/pro.4709.
  PUBMED    37347539
REFERENCE   2  (bases 1 to 4852)
  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: "Protein 
            Sci. 2023 Jun 22:e4709. doi: 10.1002/pro.4709."
COMMENT     Sequence Label: pH2B-miRFP704ano
FEATURES             Location/Qualifiers
     source          1..4852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(225..244)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    269..316
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      645..1233
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1134..1153
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1408..1711
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1712..1915
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1866..1886
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2004..2381
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     polyA_signal    2987..3108
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3024..3043)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3078..3097
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3115..3570)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3252..3273)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3464..3483
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3597..3701
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3703..4060
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3703..3894
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3726..3746)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3911..4046
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(4095..4852,1..37)
                     /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             join(4095..4852,1..37)
                     /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(4149..4168)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4759..4778
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
       61 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
      121 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
      181 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
      241 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
      301 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
      361 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
      421 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
      481 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
      541 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
      601 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
      661 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
      721 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
      781 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
      841 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
      901 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
      961 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     1021 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     1081 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     1141 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     1201 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     1261 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     1321 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
     1381 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     1441 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     1501 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     1561 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     1621 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     1681 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     1741 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     1801 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     1861 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     1921 agtgaaccgt cagatccgct agcgctaccg gactcagatc tcgagctcaa gcttcgaatt
     1981 ctgcagtcga cggtaccgcc accatgccag agccagcgaa gtctgctccc gccccgaaaa
     2041 agggctccaa gaaggcggtg actaaggcgc agaagaaagg cggcaagaag cgcaagcgca
     2101 gccgcaagga gagctattcc atctatgtgt acaaggttct gaagcaggtc caccctgaca
     2161 ccggcatttc gtccaaggcc atgggcatca tgaattcgtt tgtgaacgac attttcgagc
     2221 gcatcgcagg tgaggcttcc cgcctggcgc attacaacaa gcgctcgacc atcacctcca
     2281 gggagatcca gacggccgtg cgcctgctgc tgcctgggga gttggccaag cacgccgtgt
     2341 ccgagggtac taaggccatc accaagtaca ccagcgctaa ggatccaccg gtaatggcaa
     2401 acctggacaa gatgctgaac accatcgtga ccgaggtgcg ccagttcctg caagtggaca
     2461 gagtgtgcgt gttccagttc gaggaagatt actccggcag cgtcgtcgtg gaagccgtgg
     2521 acgacagatg gaacagcatc ctgaagaccc aggtgcggga ctgctacttc atggaaacca
     2581 gaggtgagga atacttgcac ggcagatacc aggccatcgc cgacatctac caggccaatc
     2641 tgctggagag ctacagagac ctgctgggcc agtttcaggt gcgggccatt ctggctgtcc
     2701 ccatcatcaa gggcaagaag ctgtggggcc tgctggtggc ccaccaactg gctgcccctc
     2761 ggagctggca gacctgggaa atcgagttcc tgaaacagca agccgtggtg atgggcatcg
     2821 ccatccagca gagctagctg aattctgcag atatccagca cagtggcggc cgcgactcta
     2881 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     2941 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     3001 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3061 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg
     3121 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     3181 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     3241 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3301 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     3361 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3421 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3481 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3541 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     3601 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3661 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     3721 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3781 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3841 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3901 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3961 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     4021 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     4081 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4141 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4201 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4261 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4321 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4381 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4441 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4501 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4561 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4621 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4681 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4741 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4801 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cg
//
