LOCUS       sequence-233689-        8024 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This vector encode a His6-tagged proteinA-Tnp(hyperactive form) 
            fusion protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8024)
  AUTHORS   Carter B, Ku WL, Kang JY, Hu G, Perrie J, Tang Q, Zhao K
  TITLE     Mapping histone modifications in low cell number and single cells 
            using antibody-guided chromatin tagmentation (ACT-seq).
  JOURNAL   Nat Commun. 2019 Aug 20;10(1):3747. doi: 10.1038/s41467-019-11559-1.
  PUBMED    31431618
REFERENCE   2  (bases 1 to 8024)
  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: "2019-08-20"; volume: "10(1):3747. doi"; pages: " 
            10.1038/s41467-019-11559-1"
COMMENT     Sequence Label: pET15bpATnp
FEATURES             Location/Qualifiers
     source          1..8024
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     68..87
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     171..190
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        171..189
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    190..214
                     /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)."
     RBS             229..251
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             270..287
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             297..314
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             771..1196
                     /codon_start=1
                     /product="immunoglobulin binding domains of Staphylocuccus 
                     aureus protein A"
                     /label=protein A
                     /translation="SLKDDPSQSANLLSEAKKLNESQAPKADNKFNKEQQNAFYEILHL
                     PNLNEEQRNGFIQSLKDDPSQSANLLAEAKKLNDAQAPKADNKFNKEQQNAFYEILHLP
                     NLTEEQRNGFIQSLKDDPSVSKEILAEAKKLNDAQAPK"
     primer_bind     complement(2696..2714)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2710..2757
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2932..2937,2938..2954,2955..2960))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2987..3005)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3073..3177
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3178..3246,3247..4038)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3396..3415)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4209..4797
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4698..4717
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4951..4968
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4983..5122
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5048..5067)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5208..5230
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5224..5415)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     7770..7789
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7809..7886)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
       61 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
      121 cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat taatacgact
      181 cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt
      241 taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc agcggcctgg
      301 tgccgcgcgg cagccatatg catgcgcaac acgatgaagc tcaacaaaat gctttttatc
      361 aagtcttaaa tatgcctaac ttaaatgctg atcaacgcaa tggttttatc caaagcctta
      421 aagatgatcc aagccaaagt gctaacgttt taggtgaagc taaaaaactt aatgactctc
      481 aagctccaaa agctgatgcg caacaaaata acttcaacaa agatcaacaa agcgccttct
      541 atgaaatttt gaacatgcct aacttaaacg aagcgcaacg caatggtttc attcaaagtc
      601 ttaaagacga tccaagccaa agcactaacg ttttaggtga agctaaaaaa ttaaacgaat
      661 ctcaagcacc gaaagctgac aacaatttca acaaagaaca acaaaatgct ttctatgaaa
      721 tcttgaacat gcctaacttg aacgaagaac aacgcaatgg tttcatccaa agcttaaaag
      781 atgacccaag tcaaagtgct aacctattgt cagaagctaa aaagttaaat gaatctcaag
      841 caccgaaagc ggataacaaa ttcaacaaag aacaacaaaa tgctttctat gaaatcttac
      901 atttacctaa cttaaacgaa gaacaacgca atggtttcat ccaaagctta aaagatgacc
      961 caagccaaag cgctaacctt ttagcagaag ctaaaaagct aaatgatgca caagcaccaa
     1021 aagctgacaa caaattcaac aaagaacaac aaaatgcttt ctatgaaatt ttacatttac
     1081 ctaacttaac tgaagaacaa cgtaacggct tcatccaaag ccttaaagac gatccttcag
     1141 tgagcaaaga aattttagca gaagctaaaa agctaaacga tgctcaagca ccaaaaggtg
     1201 gcggtgaatt catgataact tctgctcttc atcgtgcggc cgactgggct aaatctgtgt
     1261 tctcttcggc ggcgctgggt gatcctcgcc gtactgcccg cttggttaac gtcgccgccc
     1321 aattggcaaa atattctggt aaatcaataa ccatctcatc agagggtagt aaagccgcgc
     1381 aggaaggcgc ttaccgattt atccgcaatc ccaacgtttc tgccgaggcg atcagaaagg
     1441 ctggcgccat gcaaacagtc aagttggctc aggagtttcc cgaactgctg gccattgagg
     1501 acaccacctc tttgagttat cgccaccagg tcgccgaaga gcttggcaag ctgggctcta
     1561 ttcaggataa atcccgcgga tggtgggttc actccgttct cttgctcgag gccaccacat
     1621 tccgcaccgt aggattactg catcaggagt ggtggatgcg cccggatgac cctgccgatg
     1681 cggatgaaaa ggagagtggc aaatggctgg cagcggccgc aactagccgg ttacgcatgg
     1741 gcagcatgat gagcaacgtg attgcggtct gtgaccgcga agccgatatt catgcttatc
     1801 tgcaggacaa actggcgcat aacgagcgct tcgtggtgcg ctccaagcac ccacgcaagg
     1861 acgtagagtc tgggttgtat ctgtacgacc atctgaagaa ccaaccggag ttgggtggct
     1921 atcagatcag cattccgcaa aagggcgtgg tggataaacg cggtaaacgt aaaaatcgac
     1981 cagcccgcaa ggcgagcttg agcctgcgca gtgggcgcat cacgctaaaa caggggaata
     2041 tcacgctcaa cgcggtgctg gccgaggaga ttaacccgcc caagggtgag accccgttga
     2101 aatggttgtt gctgaccagc gaaccggtcg agtcgctagc ccaagccttg cgcgtcatcg
     2161 acatttatac ccatcgctgg cggatcgagg agttccataa ggcatggaaa accggagcag
     2221 gagccgagag gcaacgcatg gaggagccgg ataatctgga gcggatggtc tcgatcctct
     2281 cgtttgttgc ggtcaggctg ttacagctca gagaaagctt cacgccgccg caagcactca
     2341 gggcgcaagg gctgctaaag gaagcggaac acgtagaaag ccagtccgca gaaacggtgc
     2401 tgaccccgga tgaatgtcag ctactgggct atctggacaa gggaaaacgc aagcgcaaag
     2461 agaaagcagg tagcttgcag tgggcttaca tggcgatagc tagactgggc ggttttatgg
     2521 acagcaagcg aaccggaatt gccagctggg gcgccctctg ggaaggttgg gaagccctgc
     2581 aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagatcc
     2641 tttaaggatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     2701 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     2761 aaaggaggaa ctatatccgg atatcccgca agaggcccgg cagtaccggc ataaccaagc
     2821 ctatgcctac agcatccagg gtgacggtgc cgaggatgac gatgagcgca ttgttagatt
     2881 tcatacacgg tgcctgactg cgttagcaat ttaactgtga taaactaccg cattaaagct
     2941 tatcgatgat aagctgtcaa acatgagaat tcttgaagac gaaagggcct cgtgatacgc
     3001 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     3061 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3121 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3181 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3241 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3301 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3361 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3421 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3481 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3541 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     3601 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     3661 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     3721 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3781 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3841 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3901 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3961 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4021 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4081 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4141 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4201 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4261 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4321 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     4381 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4441 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4501 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4561 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4621 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4681 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4741 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4801 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4861 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     4921 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     4981 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
     5041 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     5101 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     5161 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     5221 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     5281 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     5341 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     5401 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
     5461 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     5521 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
     5581 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
     5641 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
     5701 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
     5761 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
     5821 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
     5881 tggccaggac ccaacgctgc ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc
     5941 gatggatatg ttctgccaag ggttggtttg cgcattcaca gttctccgca agaattgatt
     6001 ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc
     6061 gaggtggccc ggctccatgc accgcgacgc aacgcgggga ggcagacaag gtatagggcg
     6121 gcgcctacaa tccatgccaa cccgttccat gtgctcgccg aggcggcata aatcgccgtg
     6181 acgatcagcg gtccaatgat cgaagttagg ctggtaagag ccgcgagcga tccttgaagc
     6241 tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc gggcatcccg
     6301 atgccgccgg aagcgagaag aatcataatg gggaaggcca tccagcctcg cgtcgcgaac
     6361 gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg
     6421 ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg
     6481 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
     6541 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
     6601 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
     6661 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
     6721 ttgcgctcac tgcccgcttt ccagtcggga aactgtcgtg ccagctgcat taatgaatcg
     6781 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     6841 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     6901 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     6961 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg
     7021 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     7081 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     7141 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     7201 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     7261 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     7321 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     7381 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     7441 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     7501 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     7561 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     7621 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     7681 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     7741 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     7801 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     7861 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     7921 cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg
     7981 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggc
//
