LOCUS       sequence-368779-        5229 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genomic targeting of SNAP tag at H3f3b C-terminal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5229)
  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 5229)
  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 H3f3b
FEATURES             Location/Qualifiers
     source          1..5229
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(1543..2088)
                     /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(2995..3011)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2996..3012)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3041..3061)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3041..3059)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3080..3096)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3080..3096)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3093..3115)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3104..3120
                     /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(3128..3134,3135..3152,3153..3158))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3173..3194
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3311..3328)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3482..4070)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3562..3581)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4241..5032,5033..5101))
                     /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     4864..4883
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5102..5206)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggtggc ggccgctcta gaactaggaa ctcaatctag gcctaagacc aaacacaatg
      721 aaaagctgca ctaactagat tagtaataga tgatgctggt gtgaatagct tgttatttta
      781 ctctagagcc cttataaata aaatccccgt tagtgttgtg ttaccagcca gagggtcagg
      841 ggttagtgaa catgtggtag aatgaggact tatgcaaggt ttaatacgca tagcattttt
      901 ctactgttaa aaacaaatgc agtctagtca ggaaatacca actggactaa attacacaca
      961 ccttaatgac aagactcccc accacttgtg aatgtaaaac atttaattta aaaatgttga
     1021 acactacaat atataaaata gctattataa atgcacatag tgtattctat agctgccagg
     1081 tttacttttt tttttttaaa ggaaactgtt acactgtggc taaaacttgt atcttcaacc
     1141 tttgaaaaag cccacattct atcacagtga tgtatggtta aacacttgga tcaagtcata
     1201 accagtttta ttgcaaaagg accctgtaca catttatcaa ttctagtacc ttaatagcta
     1261 cccaacaagt cattaacata cagaaacatg catcatgaaa agcaagaagt atcacccatc
     1321 ccttctgcgt attagcaact tgtcactcct gagccacagt gctcacatca ctgaggtctg
     1381 tgaacagtca gtcactcttc ccattcatcc tgagtgaaag atggaatgac ttaagtacaa
     1441 atgcaacata ttataaacaa tttcttacaa aaaaagtcac aaattaaacc aaagtatttt
     1501 acagaattta ctacaaaatg ccataaaaac cgcttcaact tatcccagac ccggtttacc
     1561 cagacgatga ccttcatggg ccagcagcca ctctttcacc gccagaccac cttcataacc
     1621 gccaaccgca ccgctagagc taaccacacg atggcacgga atcagaatcg gcaccgggtt
     1681 accgctcagc gcggttttaa cggcggcggt ggccgccgga ttacccgcca gggccgccag
     1741 ctgctgatag ctaatcactt cgccaaattt aaccactttc agcagtttcc acagcacctg
     1801 acgggtaaag ctctcctgct gaaacaccgg atgatgcagc gccggaaccg gaaattcttc
     1861 aatcgcttcc ggctgatgaa aatacgcgtt cagccacgcg gtcgcctgca tcagcggttc
     1921 cggaccaccc agcacggcgg ccggggccgg aacttcaacc gcatcggccg cgctggtgcc
     1981 tttacccagc agtttaattt catgcaggcc ctgttcgcag ccgctcagtt ccagtttgcc
     2041 cagcgggcta tccagggtgg tacgtttcat ttcgcaatct ttgtccatgc tgccacctcc
     2101 ggatcctcct ccagctctct ccccccgtat ccggcgagcc aactggatgt ctttgggcat
     2161 gatggtgact ctcttggcgt ggatggcaca cagattggta tcttcaaaca accccaccag
     2221 gtatgcttcg ctagcctcct gttgaggaca agagcaacag cattaacact cctactggag
     2281 caatcaagcc gccgccccga gtgccaccaa gccttttgtc cctttacctg aagggcaccg
     2341 atggctgcac tttgaaacct caagtcggtt ttgaaatcct gggcgatctc cctcaccaat
     2401 ctctggaatg gcagcttccg gatgagcagc tcagtcgatt tctggtaacg acggatctct
     2461 ctcagagcca cggtccctgg cctgcagtca ccagaagaag gaaggcgtca acggtcaggg
     2521 agaagggaga gagcgcccgc ccgcaagccg gcggccaagt cattgttccc agccctctgc
     2581 ctacctgtag cggtgaggct tcttcacccc gccggtagag ggcgcgcttt tccgagccgc
     2641 cttggtggcc agctgtttgc ggggggcttt cccaccggtg gacttcctag cggtctgctt
     2701 ggttcgggcc attttttttc acctgcgaga caagcgacgt tcaaggccag gtcaccgggc
     2761 tccggctgcc aggcgcccct cctcgccgca gcagatggcg gccgccgcct cctccccctc
     2821 ccctaatgac tcaggctgcg ccgggctgcg gcctcccccg ggagggggag cgcggggagg
     2881 agcaacttcc cgctccacgg gcggccggaa acaccgcggc gggacctgtg agtcaccacc
     2941 gggaagcaga caggatggat cccccgggct gcaggaattc gatatcaagc ttatcgatac
     3001 cgtcgacctc gagggggggc ccggtaccca gcttttgttc cctttagtga gggttaattg
     3061 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     3121 ttccacacaa catacgagcc gggagcataa agtgtaaagc ctggggtgcc taatgagtga
     3181 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3241 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3301 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3361 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3421 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3481 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3541 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3601 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3661 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3721 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3781 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3841 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3901 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3961 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4021 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4081 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4141 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4201 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4261 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4321 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4381 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4441 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4501 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4561 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4621 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4681 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4741 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4801 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4861 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4921 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4981 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5041 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5101 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5161 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5221 aagtgccac
//
