LOCUS       sequence-386841-        6555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The human HBO1 is fused to C-terminal GFP of EGFP-C1  (Clontech).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6555)
  AUTHORS   Heinz KS, Rapp A, Casas-Delucchi CS, Lehmkuhl A, Romero-Fernandez I,
            Sanchez A, Kramer OH, Marchal JA, Cardoso MC
  TITLE     DNA replication dynamics of vole genome and its epigenetic 
            regulation.
  JOURNAL   Epigenetics Chromatin. 2019 Mar 14;12(1):18. doi: 
            10.1186/s13072-019-0262-0.
  PUBMED    30871586
REFERENCE   2  (bases 1 to 6555)
  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: "Epigenetics
            Chromatin."; date: "2019-03-14"; volume: "12(1):18. doi"; pages: " 
            10.1186/s13072-019-0262-0"
COMMENT     Sequence Label: pGFP-HBO1-G485A pc2201
FEATURES             Location/Qualifiers
     source          1..6555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        33..224
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     181..201
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(226..330)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      357..812
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(444..463)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     654..675
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    819..940
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(830..849)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     884..903
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(2954..3664,3665..3667,3668..3670))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGYKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2996..3017)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3345..3364
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3604..3625
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3667..3676
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3715..3918)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3744..3764)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3919..4222
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(4397..4985)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4477..4496)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5314..5361
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5386..5405
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5593..6387)
                     /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             complement(5593..6387)
                     /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(5704..5723)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6314..6333
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6422..6555,1..224))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6436..6555,1..16)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
       61 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
      121 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
      181 accctaactg acacacattc cacagctggt tctttccgcc tcaggactct tcctttttca
      241 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
      301 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc
      361 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
      421 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
      481 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
      541 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
      601 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
      661 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
      721 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
      781 gaattttaac aaaatattaa cgcttacaat ttacgcgtta agatacattg atgagtttgg
      841 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
      901 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
      961 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     1021 caaatgtggt atggctgatt atgatcagtt atctagatcc ggtggatcca gtgcccttgg
     1081 gaggggtcca ttttaagcag ctggggtcca tggttttatt ggagttggac cttttggcct
     1141 ctttggctat ccactcatca atcaggtcct gtctctttaa aactaggtgt tttcccttcc
     1201 agtatttgag catctgaagg gcttgcagag tgctgacaat gtccacagga ttcacagccg
     1261 tctcctgact gatttctttg atagaaatct ctttgccttg aaaattatgc aggtagcgga
     1321 gaagtacttc tttccagtaa ctgcgatagc ttataagccc cagatctgag agtggacgtt
     1381 ctggggagcc aactttttct tcgactttgg aaagcaaata actgaaatca ataagcatct
     1441 tgccataggc ctgtctcatg tactgaggca tagtaaggat acaggagacg ttgtagttga
     1501 ggaatgaatt cttttcctta gaaaaatatc caatcaggtg acagccagtg ttgtccgcct
     1561 ctgtcataac atagaacagg aagggctcca catcataata taatgtcttg tggtccagaa
     1621 aaagtttggc caacaggcac aggttttggc agtagatctt gtttttcttg ccatccactt
     1681 caaacacaga gattgaacct ttgcgatata tctcatcacc aggtgggtgt ttccacacac
     1741 atttggccat gtgccggcgg agtatcgttt ggctcttcat atattttaaa cagaattcac
     1801 acatatagag acgtcccagc cgtgcatatt cttcaggata tggagaatga taccaggtat
     1861 caagctcata gcggccaaaa gcaattgttt taatcatgtt gcttccctct gtgatttggc
     1921 cttgcagcct taacttctcc aaatcctctg aagcccgggc ttgtgctctt cggaaaagat
     1981 ccaagtcata ctcgcttgtc aggttttcta aaagaggttc ccgtgtgttc ccataggtct
     2041 gtctgtgttc catatatttc tctttctgtt ctttgctcag tccagaattt cttttcttcc
     2101 tgagttcagc cactttttcc ttatatcgaa gctgcctctc cgttggtgcc tggtgcctgg
     2161 ttgcatgcct gttgttgtca tcttgcctgt gagacagcat cctttcttct atctgcttat
     2221 cccggctctg tgctctcacc ttgcattcgt cagctgagag gttatgatac agtgggcatc
     2281 ctgagatgga gaaatgtctc tcatgttttc ctgtaaggtg tcctagagag ttacagcctg
     2341 gtgtaggaca cttcatattg aagttgtagc tttcatggaa gcgacggcgc ttggggcgat
     2401 gagagagatc actgcctgag tccttcaggg acatgtcctt ggcaatgctc tcatcgtgag
     2461 atacattggg gctggagata tctatgtcag actcagaaga aggcgcattt ccagttggag
     2521 ttcgaggcgg tgactcatca tgatcagctg tatttttagt ttctctatct gaaaaatcaa
     2581 ccacttgctc agtttctgaa ccagatgaac gagtctgccg aagagggtat tttttcggtg
     2641 tcactggggt aggctgctgc tgactacggg tcactcttct ggtagagtaa gcaggctcct
     2701 cagtgccaaa agactgcaga tttcgaacag gactggaatc ttgagaactc tggcttagcc
     2761 tggctgagga gcgggtgact cgagcagatc gtcgggatgt gccatcactt tctgaactgt
     2821 ctgtgtgctc gagatctgta gaaaaatcgg aatcttcggt tccatctgaa ctactgcctg
     2881 cattcctctt ccttcgcacc atggtaccgt cgactgcaga attcgaagct tgagctcgag
     2941 atctgagtcc ggacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga
     3001 actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagg gcggactggg
     3061 tgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct
     3121 ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc ttgaagttca
     3181 ccttgatgcc gttcttctgc ttgtcggcca tgatatagac gttgtggctg ttgtagttgt
     3241 actccagctt gtaccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga
     3301 tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt
     3361 cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga
     3421 agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg
     3481 tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg
     3541 tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt
     3601 ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct
     3661 tgctcaccat ggtggcgacc ggtagcgcta gcggatctga cggttcacta aaccagctct
     3721 gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt
     3781 gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc
     3841 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
     3901 ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt
     3961 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     4021 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     4081 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     4141 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     4201 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
     4261 aattgattac tattaataac taatgcatgg cggtaatacg gttatccaca gaatcagggg
     4321 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4381 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4441 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4501 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4561 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4621 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4681 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4741 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4801 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4861 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4921 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4981 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5041 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5101 aaaaatgaag ttttaaatca atctaaagta tatatgagta acctgaggct atggcagggc
     5161 ctgccgcccc gacgttggct gcgagccctg ggccttcacc cgaacttggg gggtggggtg
     5221 gggaaaagga agaaacgcgg gcgtattggc cccaatgggg tctcggtggg gtatcgacag
     5281 agtgccagcc ctgggaccga accccgcgtt tatgaacaaa cgacccaaca ccgtgcgttt
     5341 tattctgtct ttttattgcc gtcatagcgc gggttccttc cggtattgtc tccttccgtg
     5401 tttcagttag cctcccccta gggtgggcga agaactccag catgagatcc cagcgctgga
     5461 ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg
     5521 cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc
     5581 ccagagtccc gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc
     5641 gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc
     5701 agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc
     5761 acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc
     5821 gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc tggcgaacag
     5881 ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc
     5941 ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt
     6001 agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc
     6061 aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc
     6121 ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag
     6181 ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt
     6241 gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc
     6301 gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc
     6361 tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcgat
     6421 ctttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg
     6481 ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat
     6541 gggcggaact gggcg
//
