LOCUS       sequence-292022-        5304 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Histone 2B fused to monomeric near-infrared fluorescent protein 
            miRFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5304)
  AUTHORS   Piatkevich KD, Jung EE, Straub C, Linghu C, Park D, Suk HJ, Hochbaum
            DR, Goodwin D, Pnevmatikakis E, Pak N, Kawashima T, Yang CT, Rhoades
            JL, Shemesh O, Asano S, Yoon YG, Freifeld L, Saulnier JL, Riegler C,
            Engert F, Hughes T, Drobizhev M, Szabo B, Ahrens MB, Flavell SW, 
            Sabatini BL, Boyden ES
  TITLE     A robotic multidimensional directed evolution approach applied to 
            fluorescent voltage reporters.
  JOURNAL   Nat Chem Biol. 2018 Feb 26. pii: 10.1038/s41589-018-0004-9. doi: 
            10.1038/s41589-018-0004-9.
  PUBMED    29483642
REFERENCE   2  (bases 1 to 5304)
  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 Chem 
            Biol. 2018 Feb 26. pii: 10.1038/s41589-018-0004-9. doi: 
            10.1038/s41589-018-0004-9."
COMMENT     Sequence Label: pmiRFP-H2B-C1
FEATURES             Location/Qualifiers
     source          1..5304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        60..363
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        364..567
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     518..538
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      606..615
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1584..1961
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     polyA_signal    2091..2212
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2128..2147)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2182..2201
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2219..2674)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2356..2377)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2568..2587
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2701..2805
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2807..3164
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2807..2998
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2830..2850)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3015..3150
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3199..3993
                     /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             3199..3993
                     /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(3253..3272)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3863..3882
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4181..4200)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4225..4272
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4601..5189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5090..5109
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
       61 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      121 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      181 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      241 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      301 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      361 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      421 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      481 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      541 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac
      601 cggtcgccac catggtcgca ggacacgcaa gtggaagccc cgactttgga accgccgatc
      661 cgagcgactg tgagagagaa gaaatccatc tggccggatc catccagcca cacggaacac
      721 tgctggtggt ctccgagcct gaccatagaa tcattcaggc atctgccaac gccgctgaat
      781 tcctgaatct gggaagtgtg ctgggagtcc ccctggctga gatcgacggg gatctgctga
      841 tcaagattct gccccacctg gaccctaccg cagaaggaat gccagtggcc gtcaggtgcc
      901 gcattggaaa cccctccaca gagtacgatg gactgatgca tcgaccacct gaaggaggac
      961 tgatcattga gctggaacgg gctgggccac ccattgatct gagcggaaca ctggctcccg
     1021 cactggagcg aattcgaact gcaggatccc tgagggctct gtgcgacgat actgcactgc
     1081 tgttccagca gtgtaccggc tacgaccgag tgatggtcta tcggttcgat gaacaggggc
     1141 acggagaggt gtattcagaa atacatgtga ctggactgga gagctacttt ggcaacaggt
     1201 atcctagctc cctcgtgcca cagatggccc ggagactgta cgaaagacag agggtgcgcg
     1261 tcctggtgga tgtctcttat cagcctgtgc cactggagcc aagactgagt cccctgactg
     1321 gcagggacct ggatatgtca gggtgttttc tgcgctctat gagtcctact cacctgcagt
     1381 tcctgaagaa tatgggcgtg cgggcaacac tggtggtcag cctggtggtc ggaggcaaac
     1441 tgtgggggct ggtgatctgc caccattatc tgccacgctt catccatttt gagctgcgag
     1501 caatttgtga gctgctggcc gaagctatcg caacccgcat tacagccctg tccggaggcc
     1561 tcagatctgg aggctccgga ggcccagagc cagcgaagtc tgctcccgcc ccgaaaaagg
     1621 gctccaagaa ggcggtgact aaggcgcaga agaaaggcgg caagaagcgc aagcgcagcc
     1681 gcaaggagag ctattccatc tatgtgtaca aggttctgaa gcaggtccac cctgacaccg
     1741 gcatttcgtc caaggccatg ggcatcatga attcgtttgt gaacgacatt ttcgagcgca
     1801 tcgcaggtga ggcttcccgc ctggcgcatt acaacaagcg ctcgaccatc acctccaggg
     1861 agatccagac ggccgtgcgc ctgctgctgc ctggggagtt ggccaagcac gccgtgtccg
     1921 agggtactaa ggccatcacc aagtacacca gcgctaagta aggatccacc ggatctagat
     1981 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     2041 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     2101 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     2161 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
     2221 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     2281 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     2341 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     2401 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     2461 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     2521 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     2581 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     2641 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     2701 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2761 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     2821 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     2881 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     2941 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3001 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3061 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     3121 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     3181 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3241 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3301 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3361 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     3421 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3481 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3541 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3601 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3661 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3721 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3781 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3841 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     3901 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     3961 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4021 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4081 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4141 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     4201 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     4261 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     4321 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     4381 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     4441 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4501 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4561 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4621 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4681 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4741 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4801 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4861 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4921 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4981 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5041 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5101 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5161 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5221 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     5281 ataaccgtat taccgccatg catt
//
