LOCUS       sequence-387900-        5331 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the fusion protein of H2B-miRFP720 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5331)
  AUTHORS   Zhang H, Papadaki S, Sun X, Wang X, Drobizhev M, Yao L, Rehbock M, 
            Koster RW, Wu L, Namikawa K, Piatkevich KD
  TITLE     Quantitative assessment of near-infrared fluorescent proteins.
  JOURNAL   Nat Methods. 2023 Sep 4. doi: 10.1038/s41592-023-01975-z.
  PUBMED    37666982
REFERENCE   2  (bases 1 to 5331)
  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 
            Methods. 2023 Sep 4. doi: 10.1038/s41592-023-01975-z."
COMMENT     Sequence Label: pH2B-miRFP720-N1
FEATURES             Location/Qualifiers
     source          1..5331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             664..1041
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             1054..2004
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 720 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP720
                     /note="derived from Rhodopseudomonas palustris RpBphP2"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGSVIAEDRCAEVESKLGLHYPASFIPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPNHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACKRVAERLATQIGVMEE"
     polyA_signal    2126..2247
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2163..2182)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2217..2236
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2254..2709)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2391..2412)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2603..2622
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2736..2840
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2842..3199
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2842..3033
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2865..2885)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3050..3185
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3234..4028
                     /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             3234..4028
                     /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(3288..3307)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3898..3917
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4216..4235)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4260..4307
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4636..5224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5125..5144
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcc
      661 accatgccag agccagcgaa gtctgctccc gccccgaaaa agggctccaa gaaggcggtg
      721 actaaggcgc agaagaaagg cggcaagaag cgcaagcgca gccgcaagga gagctattcc
      781 atctatgtgt acaaggttct gaagcaggtc caccctgaca ccggcatttc gtccaaggcc
      841 atgggcatca tgaattcgtt tgtgaacgac attttcgagc gcatcgcagg tgaggcttcc
      901 cgcctggcgc attacaacaa gcgctcgacc atcacctcca gggagatcca gacggccgtg
      961 cgcctgctgc tgcctgggga gttggccaag cacgccgtgt ccgagggtac taaggccatc
     1021 accaagtaca ccagcgctaa ggatccaccg gtcatggcgg aaggatccgt cgccaggcag
     1081 cctgacctct tgacctgcga cgatgagccg atccatatcc ccggtgccat ccaaccgcat
     1141 ggactgctgc tcgccctcgc cgccgacatg acgatcgttg ccggcagcga caaccttccc
     1201 gaactcaccg gactggcgat cggcgccctg atcggccgct ctgcggccga tgtcttcgac
     1261 tcggagacgc acaaccgtct gacgatcgcc ttggccgagc ccggggcggc cgtcggagca
     1321 ccgatcactg tcggcttcac gatgcgaaag gacgcaggct tcatcggctc ctggcatcgc
     1381 catgatcagc tcatcttcct cgagctcgag cctccccagc gggacgtcgc cgagccgcag
     1441 gcgttcttcc gccgcaccaa cagcgccatc cgccgcctgc aggccgccga aaccttggaa
     1501 agcgcctgcg ccgccgcggc gcaagaggtg cggaagatta ccggcttcga tcgggtgatg
     1561 atctatcgct tcgcctccga cttcagcggg tccgtgatcg cagaggatcg gtgcgccgag
     1621 gtcgagtcaa aactaggcct gcactatcct gcctcattca tcccggcgca ggcccgtcgg
     1681 ctctatacca tcaacccggt acggatcatt cccgatatca attatcggcc ggtgccggtc
     1741 accccagacc tcaatccggt caccgggcgg ccgattgatc ttagcttcgc catcctgcgc
     1801 agcgtctcgc ccaaccatct ggagttcatg cgcaacatag gcatgcacgg cacgatgtcg
     1861 atctcgattt tgcgcggcga gcgactgtgg ggattgatcg tttgccatca ccgaacgccg
     1921 tactacgtcg atctcgatgg ccgccaagcc tgcaagagag tcgccgagag actggccacc
     1981 cagatcggcg tgatggaaga gtaggcggcc gcgactctag atcataatca gccataccac
     2041 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     2101 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     2161 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2221 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa
     2281 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     2341 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     2401 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     2461 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     2521 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     2581 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     2641 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     2701 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2761 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     2821 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     2881 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2941 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3001 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3061 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3121 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3181 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     3241 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3301 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3361 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     3421 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3481 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3541 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     3601 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     3661 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     3721 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     3781 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     3841 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     3901 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     3961 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4021 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4081 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4141 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4201 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     4261 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     4321 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     4381 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     4441 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     4501 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4561 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4621 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4681 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4741 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4801 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4861 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4921 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4981 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5041 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5101 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5161 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5221 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5281 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg c
//
