LOCUS       sequence-447893-        5834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of macroH2A1.1 tagged N-terminal to GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5834)
  AUTHORS   Arroyo M, Casas-Delucchi CS, Pabba MK, Prorok P, Pradhan SK, Rausch 
            C, Lehmkuhl A, Maiser A, Buschbeck M, Pasque V, Bernstein E, Luck K,
            Cardoso MC
  TITLE     Histone variant macroH2A1 regulates synchronous firing of 
            replication origins in the inactive X chromosome.
  JOURNAL   Nucleic Acids Res. 2024 Aug 27:gkae734. doi: 10.1093/nar/gkae734.
  PUBMED    39189450
REFERENCE   2  (bases 1 to 5834)
  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: "Nucleic 
            Acids Res. 2024 Aug 27:gkae734. doi: 10.1093/nar/gkae734."
COMMENT     Sequence Label: macroH2A1.1-GFP (pc2188)
FEATURES             Location/Qualifiers
     source          1..5834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(81..100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    918..965
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     990..1009
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(1197..1991)
                     /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(1197..1991)
                     /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(1308..1327)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1918..1937
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2026..2383)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2040..2175
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        2192..2383
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     2340..2360
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(2385..2489)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2516..2971
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2603..2622)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2813..2834
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2978..3099
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2989..3008)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3043..3062
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(3222..3935,3936..3938,3939..3941))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(3225..3935,3936..3938,3939..3941))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3267..3288)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3616..3635
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3875..3896
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3938..3947
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5153..5356)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5182..5202)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5357..5660
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
       61 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      421 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      601 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      661 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      721 atcaatctaa agtatatatg agtaacctga ggctatggca gggcctgccg ccccgacgtt
      781 ggctgcgagc cctgggcctt cacccgaact tggggggtgg ggtggggaaa aggaagaaac
      841 gcgggcgtat tggccccaat ggggtctcgg tggggtatcg acagagtgcc agccctggga
      901 ccgaaccccg cgtttatgaa caaacgaccc aacaccgtgc gttttattct gtctttttat
      961 tgccgtcata gcgcgggttc cttccggtat tgtctccttc cgtgtttcag ttagcctccc
     1021 cctagggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc
     1081 gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc
     1141 tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg aaccccagag tcccgctcag
     1201 aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg
     1261 taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta
     1321 gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca
     1381 gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg
     1441 agatcctcgc cgtcgggcat gctcgccttg agcctggcga acagttcggc tggcgcgagc
     1501 ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt
     1561 gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta
     1621 tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat
     1681 gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg
     1741 acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct
     1801 gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg
     1861 cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc
     1921 cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct
     1981 tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cgatctttgc aaaagcctag
     2041 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc
     2101 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     2161 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     2221 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
     2281 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     2341 ccctaactga cacacattcc acagctggtt ctttccgcct caggactctt cctttttcaa
     2401 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     2461 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg
     2521 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2581 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2641 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2701 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     2761 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2821 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     2881 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2941 aattttaaca aaatattaac gcttacaatt tacgccttaa gatacattga tgagtttgga
     3001 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     3061 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     3121 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     3181 aaatgtggta tggctgatta tgatctagag tcgcggccgc tttacttgta cagctcgtcc
     3241 atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc
     3301 tcgttggggt ctttgctcag ggcggactgg gtgctcaggt agtggttgtc gggcagcagc
     3361 acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg
     3421 tcctcgatgt tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgtcggcc
     3481 atgatataga cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg
     3541 tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac
     3601 ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg
     3661 acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag
     3721 cggctgaagc actgcacgcc gtaggtcagg gtggtcacga gggtgggcca gggcacgggc
     3781 agcttgccgg tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg
     3841 ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg
     3901 ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca tggtggcgac cggtggatcc
     3961 cgggcccgcg gtaccgtcga ctgcagaatt cgaagcttgt tggcatccag tttggccatt
     4021 tcctgcacgt agatgcctat gctctcactg tcaaaaagca caaagtacac agttttgatg
     4081 gaggacgaca tagtggacac aaagtagctg gagatggcct tcagaatgag ttgggcggct
     4141 gtctgcttcg ggaacccgtt cctgccactg ccaatagacg ggaaggcgat ggacttcagc
     4201 tttctgtcat cagctagagc caggcagttt ttcaccgtct tttctagaag ttcttcacat
     4261 ttgtctgcgc cccagacagg actgttacag tggatcacaa acttggcagg caggccgtgg
     4321 cctgcactaa cagcagctcc agctacttcc aagggcccgt tctttttccg aagttccaga
     4381 acagcttcta caaactcctt gccacctttc ttctccagtg tgcttcctac ttcaccaccg
     4441 atgtagaagt cagtgtttgt cgggtgaacg acagcatcac tgtcgatcga ggcaatgtca
     4501 gcctgaacaa cttgcaactt ctggccgagg aagaggctct tggtagagag gacagtaaag
     4561 ccgtctgtag gcgcgccctc cgttgtactg tctgcgctgg ctgccttgct gacttctccc
     4621 tgcttcttag acttccgggc ccctttcttg cctcctgttt tcttagccac tggcttcttc
     4681 tgggatggag acttggcctt tttggctggt ggaggcgtga tgatggcttc caattttccc
     4741 ttggatcctc gtttcttcgc tagcaactca ggatggatgt tcggcaacac gcccccactg
     4801 gctatggtga ctccttttag cagctggttt agctcctcat cattggccac tgctaacagg
     4861 atgtgccgcg gcgtgacccg ccccttcttg ttgtctctcg ctgcattgcc ggccagctcc
     4921 aggatctcag cggtcaggta ttccagtacg gcagccatgt acacaggcgc ccccactccg
     4981 atccgatact tagggtggcc tttcttgatg taccgcagca tccgtcccac ggggaagatg
     5041 accccggcct tggctgaccg ggaggtcttg gtggatttct tcttcccacc acggctcgac
     5101 atctcgagat ctgagtccgg tagcgctagc ggatctgacg gttcactaaa ccagctctgc
     5161 ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt
     5221 tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa
     5281 tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact
     5341 gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
     5401 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
     5461 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
     5521 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     5581 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     5641 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
     5701 ttgattacta ttaataacta atgcatggcg gtaatacggt tatccacaga atcaggggat
     5761 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5821 gcgttgctgg cgtt
//
