LOCUS       sequence-259980-        5453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing SNAP-tagged histone H3.1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5453)
  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 5453)
  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-H3 Zeo
FEATURES             Location/Qualifiers
     source          1..5453
                     /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             969..1514
                     /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"
     misc_feature    1965..2538
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2132..2149)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2359..2378
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2574..2948
                     /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    3073..3194
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3110..3129)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3296..3312)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3296..3312)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3309..3331)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3320..3336
                     /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(3344..3350,3351..3368,3369..3374))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3389..3410
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3527..3544)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3698..4286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3778..3797)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4457..5248,5249..5317))
                     /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     5080..5099
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5318..5422)
                     /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 cgtcgctagc gatatcggcg cgccagcatt taaatctgta cagaccggtg
      961 aattcaccat ggacaaagac tgcgaaatga agcgcaccac cctggatagc cctctgggca
     1021 agctggaact gtctgggtgc gaacagggcc tgcaccgtat catcttcctg ggcaaaggaa
     1081 catctgccgc cgacgccgtg gaagtgcctg ccccagccgc cgtgctgggc ggaccagagc
     1141 cactgatgca ggccaccgcc tggctcaacg cctactttca ccagcctgag gccatcgagg
     1201 agttccctgt gccagccctg caccacccag tgttccagca ggagagcttt acccgccagg
     1261 tgctgtggaa actgctgaaa gtggtgaagt tcggagaggt catcagctac agccacctgg
     1321 ccgccctggc cggcaatccc gccgccaccg ccgccgtgaa aaccgccctg agcggaaatc
     1381 ccgtgcccat tctgatcccc tgccaccggg tggtgcaggg cgacctggac gtggggggct
     1441 acgagggcgg gctcgccgtg aaagagtggc tgctggccca cgagggccac agactgggca
     1501 agcctgggct gggtcctgca ggcggatccg cgtttaaact cgaggctcgc actaagcaaa
     1561 ctgctcggaa gtctactggt ggcaaggcgc cacgcaaaca gttggccact aaggcagccc
     1621 gcaaaagcgc tccggccacc ggcggcgtga aaaagcccca ccgctaccgg ccgggcaccg
     1681 tggctctgcg cgagatccgc cgttatcaga agtccactga actgcttatt cgtaaactac
     1741 ctttccagcg cctggtgcgc gagattgcgc aggactttaa aacagacctg cgtttccaga
     1801 gctccgctgt gatggctctg caggaggcgt gcgaggccta cttggtaggg ctatttgagg
     1861 acactaacct gtgcgccatc cacgccaagc gcgtcactat catgcccaag gacatccagc
     1921 tcgcccgccg catccgcgga gagagggcgt gactctcccc cccccccttt tccctctccc
     1981 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
     2041 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
     2101 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg
     2161 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
     2221 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
     2281 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
     2341 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
     2401 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt
     2461 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
     2521 tgaaaaacac gatgataagc ttgccacaac ccggtataat tcctgcagcc aatatggcca
     2581 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
     2641 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc
     2701 gggacgacgt gaccctgttc atcagcgcgg tccaggacca gatggtgccg gacaacaccc
     2761 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt
     2821 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg
     2881 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc
     2941 aggactgaca cgtgctctag ataactgatc ataatcagcc ataccacatt tgtagaggtt
     3001 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     3061 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     3121 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     3181 atcaatgtat cttaacgcgt cgagtgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3241 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     3301 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3361 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3421 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3481 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3541 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3601 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3661 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3721 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3781 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3841 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3901 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3961 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4021 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4081 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4141 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4201 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4261 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4321 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4381 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4441 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4501 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4561 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4621 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4681 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4741 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4801 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4861 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4921 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4981 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5041 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5101 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5161 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5221 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5281 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5341 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5401 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
