LOCUS       sequence-259981-        6501 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing SNAP-tagged histone macroH2A1.2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6501)
  AUTHORS   Sato H, Wu B, Delahaye F, Singer RH, Greally JM
  TITLE     Retargeting of macroH2A following mitosis to cytogenetic-scale 
            heterochromatic domains.
  JOURNAL   J Cell Biol. 2019 Jun 3;218(6):1810-1823. doi: 
            10.1083/jcb.201811109. Epub 2019 May 20.
  PUBMED    31110057
REFERENCE   2  (bases 1 to 6501)
  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"; doi: 
            "10.1083/jcb.201811109"; journalName: "J Cell Biol"; date: 
            "2019-06-3- 3"; volume: "218"; issue: "6"; pages: "1810-1823"
COMMENT     Sequence Label: pSNAP-macroH2A1.2 Zeo
FEATURES             Location/Qualifiers
     source          1..6501
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2055..2600
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     intron          2711..2940
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2923..2942
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(3013..3587,3588..3590,3591..3599)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     3407..3426
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3622..3996
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQMVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4121..4242
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4158..4177)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4344..4360)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4344..4360)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4357..4379)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4368..4384
                     /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(4392..4398,4399..4416,4417..4422))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4437..4458
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4575..4592)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4746..5334)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4826..4845)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5505..6296,6297..6365))
                     /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     6128..6147
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6366..6470)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cgatatcgaa ttgatgtcga gccgcggcgg gaagaagaaa tccaccaaga
      961 cctcccggtc agccaaggcc ggagtcatct tccctgtggg acgcatgctt cggtacatca
     1021 agaaaggcca ccctaagtat aggatcggag tgggggcacc tgtgtacatg gctgctgtcc
     1081 tggagtacct gactgctgag atcctggagc tggctggcaa tgcagcaaga gacaacaaga
     1141 agggacgggt cacaccccgg cacatcctgt tagctgtggc caatgatgaa gagctaaacc
     1201 agctgctaaa gggtgtcacc atagtcagcg ggggcgtgtt gccgaatatc catcctgagt
     1261 tgctagcgaa gaagcgagga tccaagggaa aattggaagc catcatcacg cctccgccgg
     1321 ccaaaaaggc caagtctcca tcccagaaga agccagtggc taagaagaca ggaggcaaga
     1381 aaggggcccg gaagtctaag aagaagcagg gagaagtgag caaggcggcc agcgcagaca
     1441 gtacgacgga gggcacacct acagacggct tcactgtcct ctccaccaag agcctcttcc
     1501 tcggccagaa gctgaacctt attcacagtg aaatcagtaa tttagccggc tttgaggtgg
     1561 aggccataat caatcctacc aatgctgaca ttgaccttaa agatgaccta ggaaacacac
     1621 tggagaagaa gggcggcaag gagtttgtag aagctgttct ggaactccgg aaaaagaacg
     1681 ggcccttgga ggtagctgga gctgctatta gtgcaggcca tggcctgcct gccaagtttg
     1741 tgatccactg taatagtcct gtctggggtg cagacaaatg tgaagaactt ctagaaaaga
     1801 cggtgaaaaa ctgcttggct ctagctgatg acagaaagct gaaatccatc gccttcccat
     1861 ccattggcag cggcaggaac gggttcccga agcagacagc ggcccagctc attctgaagg
     1921 ccatctccag ctactttgtc tccacgatgt cctcctccat caaaactgtg tacttcatgc
     1981 tttttgacag tgagagcata ggtatctatg tgcaggaaat ggccaagctg gacgccaaca
     2041 ccggtgaatt caccatggac aaagactgcg aaatgaagcg caccaccctg gatagccctc
     2101 tgggcaagct ggaactgtct gggtgcgaac agggcctgca ccgtatcatc ttcctgggca
     2161 aaggaacatc tgccgccgac gccgtggaag tgcctgcccc agccgccgtg ctgggcggac
     2221 cagagccact gatgcaggcc accgcctggc tcaacgccta ctttcaccag cctgaggcca
     2281 tcgaggagtt ccctgtgcca gccctgcacc acccagtgtt ccagcaggag agctttaccc
     2341 gccaggtgct gtggaaactg ctgaaagtgg tgaagttcgg agaggtcatc agctacagcc
     2401 acctggccgc cctggccggc aatcccgccg ccaccgccgc cgtgaaaacc gccctgagcg
     2461 gaaatcccgt gcccattctg atcccctgcc accgggtggt gcagggcgac ctggacgtgg
     2521 ggggctacga gggcgggctc gccgtgaaag agtggctgct ggcccacgag ggccacagac
     2581 tgggcaagcc tgggctgggt cctgcaggcg gatccgcgtt taaactcgag gttaattaat
     2641 gagcggccgc atagataact gatccagtgt gctggaatta attcgctgtc tgcgagggcc
     2701 agctgttggg gtgagtactc cctctcaaaa gcgggcatga cttctgcgct aagattgtca
     2761 gtttccaaaa acgaggagga tttgatattc acctggcccg cggtgatgcc tttgagggtg
     2821 gccgcgtcca tctggtcaga aaagacaatc tttttgttgt caagcttgag gtgtggcagg
     2881 cttgagatct ggccatacac ttgagtgaca atgacatcca ctttgccttt ctctccacag
     2941 gtgtccactc ccaggtccaa ctgcaggtcg agcatgcatc tagggcggcc aattccgccc
     3001 ctctcccccc cccccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga
     3061 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     3121 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     3181 tctcgccaaa gtaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     3241 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     3301 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     3361 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     3421 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     3481 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     3541 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataagctt gccacaaccc
     3601 ggtataattc ctgcagccaa tatggccaag ttgaccagtg ccgttccggt gctcaccgcg
     3661 cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc
     3721 gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc
     3781 caggaccaga tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag
     3841 ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc
     3901 atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc
     3961 aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctctagat aactgatcat
     4021 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     4081 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     4141 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4201 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtcg agtgcattct
     4261 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     4321 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4381 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4441 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4501 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4561 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4621 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4681 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4741 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4801 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4861 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4921 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4981 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5041 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5101 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5161 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5221 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5281 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5341 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5401 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5461 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5521 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5581 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5641 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5701 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5761 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5821 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5881 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5941 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6001 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6061 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6121 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6181 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6241 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6301 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6361 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6421 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6481 cgaaaagtgc cacctgacgt c
//
