LOCUS       sequence-368931-        5178 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genomic targeting of SNAP tag at Hist1h3g C-terminal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5178)
  AUTHORS   Xu X, Duan S, Hua X, Li Z, He R, Zhaang Z
  TITLE     Stable inheritance of H3.3-containing nucleosomes during mitotic 
            cell divisions.
  JOURNAL   Nat Commun. 2022 May 6;13(1):2514. doi: 10.1038/s41467-022-30298-4.
  PUBMED    35523900
REFERENCE   2  (bases 1 to 5178)
  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: "Nat 
            Commun."; date: "2022-05-6"; volume: "13(1):2514. doi"; pages: " 
            10.1038/s41467-022-30298-4"
COMMENT     Sequence Label: pBlueScript II SK-SNAP donor for Hist1h3g
FEATURES             Location/Qualifiers
     source          1..5178
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(28..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     350..369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     583..605
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     597..614
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     598..614
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     624..643
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        624..642
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(1499..2044)
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="optimized for bacterial expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     primer_bind     complement(2942..2958)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2943..2959)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2988..3008)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2988..3006)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3027..3043)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3027..3043)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3040..3062)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3051..3067
                     /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(3075..3081,3082..3099,3100..3105))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3120..3141
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3258..3275)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3429..4017)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3509..3528)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4188..4979,4980..5048))
                     /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     4811..4830
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5049..5153)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
      541 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
      601 aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg gagctccacc
      661 gcggtggcgg ccgctctaga actagttttc agcatccaga gttcgaactg tcttccatta
      721 tagtcactag cacatctcct aaagaccagc cttcatattt ccacaccata caccactgtg
      781 ataatgacct aggacaaatc cagtgcccaa atggctgtaa ttttcgaggt gatgattatt
      841 aagccaagtg aaacaagtga cgaaagagcc ataaagattc aggatctcaa aaagcccaaa
      901 ggagggcaag acatatacac cttaatccca gcatacagta ggaaggagaa acagatctcc
      961 caaatacaga tcatcctgat ctacaaagaa agttgcagac cagcctaagc tatgagtcca
     1021 cttaaaaaga gggtggggga ggtctattag tgggtaggga ggagactaag cacgctataa
     1081 agtgaatggt aaatccctgt aaccctttca agactggaag atcagttaat ttttctgagt
     1141 tacaatatta gtggttctgt ccttcagaaa gttcacaatt ccaaagtaaa catctgcaac
     1201 taataatttt atttttaaat tcggtaaatg cccaaattac tgtaaccatg caaagttttc
     1261 caatctttat gctttgctaa ctaagagtac aatttccatg aaaaagggta aagatatcac
     1321 ttaccaaaat attgcttgct tataatagat cggtgtgatg aaaggagacc tagcactttc
     1381 acaaatgaaa ctgattaata cccacaagtt ggaaaaagcg ctacagttct ttttttggaa
     1441 gatgggtggc tctaagaaga gcctttaaag tggttgtgtg gattaacaga aacccttatc
     1501 ccagacccgg tttacccaga cgatgacctt catgggccag cagccactct ttcaccgcca
     1561 gaccaccttc ataaccgcca accgcaccgc tagagctaac cacacgatgg cacggaatca
     1621 gaatcggcac cgggttaccg ctcagcgcgg ttttaacggc ggcggtggcc gccggattac
     1681 ccgccagggc cgccagctgc tgatagctaa tcacttcgcc aaatttaacc actttcagca
     1741 gtttccacag cacctgacgg gtaaagctct cctgctgaaa caccggatga tgcagcgccg
     1801 gaaccggaaa ttcttcaatc gcttccggct gatgaaaata cgcgttcagc cacgcggtcg
     1861 cctgcatcag cggttccgga ccacccagca cggcggccgg ggccggaact tcaaccgcat
     1921 cggccgcgct ggtgccttta cccagcagtt taatttcatg caggccctgt tcgcagccgc
     1981 tcagttccag tttgcccagc gggctatcca gggtggtacg tttcatttcg caatctttgt
     2041 ccatgctgcc acctccggat cctcctccag ccctctcccc gcggatgcga cgggccagct
     2101 ggatgtcctt gggcatgatg gtgacacgct tggcgtggat ggcgcacagg ttggtgtcct
     2161 caaacagacc cacgaggtag gcctcacagg cctcctgcag agccatgacg gccgagctct
     2221 ggaagcgcag gtcggtcttg aagtcctgcg cgatctcgcg caccaggcgc tggaacggca
     2281 gcttgcggat cagcagctcg gtcgacttct ggtagcgccg gatctcgcgc agcgccacag
     2341 tgccgggacg gtagcggtga ggcttcttca cgccgccggt ggccggggcg ctcttgcggg
     2401 cggccttggt ggccagctgc ttgcgcgggg ccttgccgcc ggtagacttg cgagcggtct
     2461 gcttagtacg agccatggca agtaaccgta gggaaaagtg agatacctaa gtagcgcact
     2521 ttggccttta agtacgcttg cgttcgcaca cgcgcaacac acgtacccgc gctcgtaccc
     2581 ctccttccca ccgagaagca tcctgatagg cgtgtctgct tttcaagtgt cccgcgcagt
     2641 agtactattg gctgaggaat tcaagtggtg attggttaac tcctgcacac ccgattggtt
     2701 aactcctgca cacagtctcc tgctagtaca gactgctaca ctcatgtttt agtctggcac
     2761 acccgagctt tcttaaattt atgtttccat ccttgtatcg agtcgcttaa agaatttgag
     2821 gcaaagaaat cttgaatttg aaatctcatg accctagata tccacaaacc actctggaaa
     2881 atatgttctc tacccgcctt tgtggatccc ccgggctgca ggaattcgat atcaagctta
     2941 tcgataccgt cgacctcgag ggggggcccg gtacccagct tttgttccct ttagtgaggg
     3001 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     3061 ctcacaattc cacacaacat acgagccggg agcataaagt gtaaagcctg gggtgcctaa
     3121 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3181 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     3241 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3301 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3361 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3421 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3481 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3541 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3601 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3661 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3721 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3781 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3841 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3901 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3961 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4021 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4081 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4141 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4201 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4261 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4321 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4381 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4441 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4501 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4561 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4621 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4681 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4741 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4801 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4861 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4921 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4981 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5041 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5101 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5161 ccccgaaaag tgccacct
//
