LOCUS       sequence-159436-        5106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Nucleus/Histones, Excitation: 383, Emission: 448.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5106)
  TITLE     Davidson Blue Fluorescent Proteins
REFERENCE   2  (bases 1 to 5106)
  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"
COMMENT     Sequence Label: EBFP2-H2B-6
FEATURES             Location/Qualifiers
     source          1..5106
                     /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"
     regulatory      1054..1063
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1060..1062,1063..1065,1066..1779)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(1105..1126)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    1901..2022
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1938..1957)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1992..2011
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2029..2484)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2166..2187)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2378..2397
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2511..2615
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2617..2974
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2617..2808
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2640..2660)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2825..2960
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3009..3803
                     /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             3009..3803
                     /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(3063..3082)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3673..3692
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3991..4010)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4035..4082
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4411..4999
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4900..4919
                     /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 gtcgccacca tggtgagcaa gggcgaggag
     1081 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1141 ttcagcgtga ggggcgaggg cgagggcgat gccaccaacg gcaagctgac cctgaagttc
     1201 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgagccac
     1261 ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1321 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcacctac
     1381 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1441 ggcgtcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caacttcaac
     1501 agccacaaca tctatatcat ggccgtcaag cagaagaacg gcatcaaggt gaacttcaag
     1561 atccgccaca acgtggagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     1621 cccatcggcg acggccccgt gctgctgccc gacagccact acctgagcac ccagtccgtg
     1681 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt ccgcaccgcc
     1741 gccgggatca ctctcggcat ggacgagctg tacaagtaag cggccgcgac tctagatcat
     1801 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     1861 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     1921 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     1981 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     2041 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2101 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2161 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2221 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     2281 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2341 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     2401 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     2461 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2521 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2581 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     2641 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2701 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2761 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     2821 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     2881 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     2941 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     3001 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3061 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3121 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3181 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3241 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3301 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3361 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3421 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3481 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     3541 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3601 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3661 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     3721 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     3781 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     3841 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     3901 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     3961 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     4021 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     4081 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     4141 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     4201 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     4261 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4321 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4381 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4441 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4501 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4561 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4621 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4681 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4741 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4801 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4861 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4921 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4981 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5041 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5101 taccgc
//
