LOCUS       sequence-447925-        5843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of macroH2A2 tagged N-terminal to GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5843)
  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 5843)
  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: macroH2A2-GFP (pc2191)
FEATURES             Location/Qualifiers
     source          1..5843
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1222..1242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2486..2495
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2492..2494,2495..2497,2498..3211)
                     /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             join(2492..2494,2495..2497,2498..3208)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2537..2558)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2798..2817)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3145..3166
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3334..3455
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3371..3390)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3425..3444
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3462..3917)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3599..3620)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3811..3830
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3944..4048
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4050..4407
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4050..4241
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4073..4093)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4258..4393
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4442..5236
                     /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             4442..5236
                     /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(4496..4515)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5106..5125
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5424..5443)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5468..5515
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact cagatctcga
     1321 gctcaagctt atgtcgggcc ggagtgggaa gaagaaaatg tccaagctgt cccgttcagc
     1381 tagggcaggt gtcatctttc cagtggggag gctgatgcgt tatctgaaga aagggacgtt
     1441 caagtaccgg atcagcgtgg gcgcccctgt ctacatggcg gcagtcattg agtacctggc
     1501 agcggaaatt ctagaattgg ccggcaatgc cgcgagggac aacaagaagg cccggatagc
     1561 cccgagacac atcttgctgg cagttgccaa tgacgaggag ctcaaccagc tgctaaaagg
     1621 agtgaccatc gccagtggag gcgtcctgcc cagaattcac cccgaactgc tggccaaaaa
     1681 gcgagggacc aaaggcaagt cggaaacgat cctctcccca cccccagaga aaagaggcag
     1741 gaaggccacg tcaggcaaga agggggggaa gaaatccaag gctgccaaac cacggacgtc
     1801 caaaaagtcc aaaccaaagg acagcgataa agaaggaact tcaaattcca cctctgaaga
     1861 tgggccaggg gatggattca ccattctgtc ttctaagagc cttgttctgg gacagaagct
     1921 gtccttaacc cagagtgaca tcagccatat tggctccatg agagtggagg gcattgtcca
     1981 cccaaccaca gccgaaattg acctcaaaga agatataggt aaagccttgg aaaaggctgg
     2041 gggaaaagag ttcttggaaa cggtaaagga gcttcgcaaa tcccaaggcc ctttggaagt
     2101 cgccgaagcc gccgtcagcc aatccagtgg actcgcagcc aaatttgtca tccactgtca
     2161 catccctcag tggggctccg acaaatgtga agaacagctt gaagagacca tcaaaaactg
     2221 cctgtcagcg gcggaggaca agaagctaaa gtccgtcgcg ttcccgcctt tccccagcgg
     2281 cagaaactgc tttcccaaac agactgcggc ccaggtgacc ctcaaagcca tctcagccca
     2341 ctttgatgac tcgagcgcgt cctcgctgaa gaacgtgtac ttcctgctct tcgacagcga
     2401 gagcatcggc atctacgtgc aggagatggc caagctcgac gccaagctgc agtcgacggt
     2461 accgcgggcc cgggatccac cggtcgccac catggtgagc aagggcgagg agctgttcac
     2521 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     2581 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     2641 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     2701 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     2761 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     2821 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2881 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     2941 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     3001 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     3061 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     3121 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     3181 cactctcggc atggacgagc tgtacaagta aagcggccgc gactctagat cataatcagc
     3241 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     3301 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     3361 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3421 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     3481 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     3541 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3601 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3661 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     3721 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     3781 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     3841 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3901 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3961 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4021 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     4081 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4141 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4201 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4261 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4321 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4381 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     4441 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4501 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4561 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4621 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4681 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4741 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4801 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4861 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4921 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     4981 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5041 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5101 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5161 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5221 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5281 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5341 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5401 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     5461 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     5521 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     5581 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     5641 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     5701 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5761 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5821 gtagaaaaga tcaaaggatc ttc
//
