LOCUS       sequence-242042-        6978 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cyan color variant of bright luminescent protein, fused to histone 
            2B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6978)
  AUTHORS   Suzuki K, Kimura T, Shinoda H, Bai G, Daniels MJ, Arai Y, Nakano M, 
            Nagai T
  TITLE     Five colour variants of bright luminescent protein for real-time 
            multicolour bioimaging.
  JOURNAL   Nat Commun. 2016 Dec 14;7:13718. doi: 10.1038/ncomms13718.
  PUBMED    27966527
REFERENCE   2  (bases 1 to 6978)
  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: "2016-12-14"; volume: "7:13718. doi"; pages: " 
            10.1038/ncomms13718"
COMMENT     Sequence Label: CeNL-H2B_pcDNA3
FEATURES             Location/Qualifiers
     source          1..6978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        145..524
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        525..728
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     679..699
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     773..792
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        773..791
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      824..833
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(830..832,833..835,836..1348)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(875..896)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1136..1155)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1483..1504
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(2441..2459)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2442..2459)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2479..2496)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2485..2709
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2755..3183
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2842..2861)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3052..3073
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3192..3212)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3197..3526
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3377..3512
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3439..3458
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3593..4387
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3647..3666)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4257..4276
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4561..4682
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4598..4617)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4652..4671
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4731..4747)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4731..4747)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4744..4766)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4755..4771
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4779..4785,4786..4803,4804..4809))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4824..4845
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4962..4979)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5133..5721)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5213..5232)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5892..6683,6684..6752))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6515..6534
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6753..6857)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6932..6951)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
       61 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
      121 atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa
      181 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      241 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      301 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      361 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      421 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      481 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      541 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      601 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      661 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      721 gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga
      781 ctcactatag ggagacccaa gcttggtacc gagctcggat cccgccacca tggtgagcaa
      841 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
      901 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
      961 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1021 cctgtcctgg ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     1081 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1141 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1201 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1261 caactacttt agcgacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
     1321 caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca
     1381 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     1441 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1501 cgtgaccgcc gccgggttgc atacactcga agatttcgtt ggggactggc gacagacagc
     1561 cggctacaac ctggaccaag tccttgaaca gggaggtgtg tccagtttgt ttcagaatct
     1621 cggggtgtcc gtaactccga tccaaaggat tgtcctgagc ggtgaaaatg ggctgaagat
     1681 cgacatccat gtcatcatcc cgtatgaagg tctgagcggc gaccaaatgg gccagatcga
     1741 aaaaattttt aaggtggtgt accctgtgga tgatcatcac tttaaggtga tcctgcacta
     1801 tggcacactg gtaatcgacg gggttacgcc gaacatgatc gactatttcg gacggccgta
     1861 tgaaggcatc gccgtgttcg acggcaaaaa gatcactgta acagggaccc tgtggaacgg
     1921 caacaaaatt atcgacgagc gcctgatcaa ccccgacggc tccctgctgt tccgagtaac
     1981 catcaacgga gtgaccggct ggcggctgtg cgaacgcatt ctggcggaat tcatgccgga
     2041 gccagcgaag tctgctcccg cgcccaagaa gggctcgaag aaagccgtga ctaaggcgca
     2101 gaagaaggac ggcaagaagc gcaagcgcag ccgcaaggag agctactccg tatacgtgta
     2161 caaggtgctg aagcaggtcc accccgacac cggcatctcc tctaaggcca tgggaatcat
     2221 gaactccttc gtcaacgaca tcttcgaacg catcgcgggt gaggcttccc gcctggcgca
     2281 ttacaacaag cgctcgacca tcacctccag ggagatccag acggccgtgc gcctgctgct
     2341 gcccggggag ttggccaagc acgccgtgtc cgagggcacc aaggccgtca ccaagtacac
     2401 cagcgctaag taactcgagc atgcatctag agggccctat tctatagtgt cacctaaatg
     2461 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2521 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2581 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2641 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     2701 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     2761 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2821 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2881 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2941 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3001 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3061 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3121 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3181 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3241 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3301 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3361 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3421 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     3481 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     3541 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     3601 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3661 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3721 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     3781 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     3841 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     3901 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     3961 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4021 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4081 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     4141 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4201 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4261 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4321 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4381 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4441 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4501 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4561 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4621 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4681 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4741 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4801 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4861 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4921 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4981 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5041 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5101 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5161 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5221 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5281 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5341 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5401 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5461 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5521 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5581 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5641 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5701 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5761 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5821 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5881 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5941 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6001 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6061 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6121 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6181 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6241 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6301 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6361 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6421 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6481 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6541 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6601 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6661 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6721 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6781 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6841 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     6901 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     6961 gccagtatct gctccctg
//
