LOCUS       sequence-230305-        8028 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 8028)
  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 8028)
  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..8028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(630..1712)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1674..1693
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1713..1790)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1996..2015
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2099..2118
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2099..2117
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2118..2142
                     /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             2157..2179
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2198..2215
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2225..2242
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2699..3124
                     /codon_start=1
                     /product="immunoglobulin binding domains of Staphylocuccus 
                     aureus protein A"
                     /label=protein A
                     /translation="SLKDDPSQSANLLSEAKKLNESQAPKADNKFNKEQQNAFYEILHL
                     PNLNEEQRNGFIQSLKDDPSQSANLLAEAKKLNDAQAPKADNKFNKEQQNAFYEILHLP
                     NLTEEQRNGFIQSLKDDPSVSKEILAEAKKLNDAQAPK"
     primer_bind     complement(4624..4642)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4638..4685
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(4860..4865,4866..4882,4883..4888))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(4915..4933)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5001..5105
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5106..5174,5175..5966)
                     /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(5324..5343)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6137..6725
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6626..6645
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6879..6896
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6911..7053
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6979..6998)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7139..7161
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7155..7346)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg
       61 ccgaggcggc ataaatcgcc gtgacgatca gcggtccagt gatcgaagtt aggctggtaa
      121 gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca
      181 tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg
      241 ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc
      301 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
      361 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
      421 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
      481 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
      541 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
      601 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
      661 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
      721 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
      781 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
      841 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
      901 actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
      961 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     1021 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     1081 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     1141 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     1201 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     1261 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     1321 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     1381 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     1441 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     1501 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     1561 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     1621 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     1681 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     1741 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     1801 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     1861 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     1921 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     1981 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     2041 ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaatta
     2101 atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga aataattttg
     2161 tttaacttta agaaggagat ataccatggg cagcagccat catcatcatc atcacagcag
     2221 cggcctggtg ccgcgcggca gccatatgca tgcgcaacac gatgaagctc aacaaaatgc
     2281 tttttatcaa gtcttaaata tgcctaactt aaatgctgat caacgcaatg gttttatcca
     2341 aagccttaaa gatgatccaa gccaaagtgc taacgtttta ggtgaagcta aaaaacttaa
     2401 tgactctcaa gctccaaaag ctgatgcgca acaaaataac ttcaacaaag atcaacaaag
     2461 cgccttctat gaaattttga acatgcctaa cttaaacgaa gcgcaacgca atggtttcat
     2521 tcaaagtctt aaagacgatc caagccaaag cactaacgtt ttaggtgaag ctaaaaaatt
     2581 aaacgaatct caagcaccga aagctgacaa caatttcaac aaagaacaac aaaatgcttt
     2641 ctatgaaatc ttgaacatgc ctaacttgaa cgaagaacaa cgcaatggtt tcatccaaag
     2701 cttaaaagat gacccaagtc aaagtgctaa cctattgtca gaagctaaaa agttaaatga
     2761 atctcaagca ccgaaagcgg ataacaaatt caacaaagaa caacaaaatg ctttctatga
     2821 aatcttacat ttacctaact taaacgaaga acaacgcaat ggtttcatcc aaagcttaaa
     2881 agatgaccca agccaaagcg ctaacctttt agcagaagct aaaaagctaa atgatgcaca
     2941 agcaccaaaa gctgacaaca aattcaacaa agaacaacaa aatgctttct atgaaatttt
     3001 acatttacct aacttaactg aagaacaacg taacggcttc atccaaagcc ttaaagacga
     3061 tccttcagtg agcaaagaaa ttttagcaga agctaaaaag ctaaacgatg ctcaagcacc
     3121 aaaaggtggc ggtgaattca tgataacttc tgctcttcat cgtgcggccg actgggctaa
     3181 atctgtgttc tcttcggcgg cgctgggtga tcctcgccgt actgcccgct tggttaacgt
     3241 cgccgcccaa ttggcaaaat attctggtaa atcaataacc atctcatcag agggtagtaa
     3301 agccgcgcag gaaggcgctt accgatttat ccgcaatccc aacgtttctg ccgaggcgat
     3361 cagaaaggct ggcgccatgc aaacagtcaa gttggctcag gagtttcccg aactgctggc
     3421 cattgaggac accacctctt tgagttatcg ccaccaggtc gccgaagagc ttggcaagct
     3481 gggctctatt caggataaat cccgcggatg gtgggttcac tccgttctct tgctcgaggc
     3541 caccacattc cgcaccgtag gattactgca tcaggagtgg tggatgcgcc cggatgaccc
     3601 tgccgatgcg gatgaaaagg agagtggcaa atggctggca gcggccgcaa ctagccggtt
     3661 acgcatgggc agcatgatga gcaacgtgat tgcggtctgt gaccgcgaag ccgatattca
     3721 tgcttatctg caggacaaac tggcgcataa cgagcgcttc gtggtgcgct ccaagcaccc
     3781 acgcaaggac gtagagtctg ggttgtatct gtacgaccat ctgaagaacc aaccggagtt
     3841 gggtggctat cagatcagca ttccgcaaaa gggcgtggtg gataaacgcg gtaaacgtaa
     3901 aaatcgacca gcccgcaagg cgagcttgag cctgcgcagt gggcgcatca cgctaaaaca
     3961 ggggaatatc acgctcaacg cggtgctggc cgaggagatt aacccgccca agggtgagac
     4021 cccgttgaaa tggttgttgc tgaccagcga accggtcgag tcgctagccc aagccttgcg
     4081 cgtcatcgac atttataccc atcgctggcg gatcgaggag ttccataagg catggaaaac
     4141 cggagcagga gccgagaggc aacgcatgga ggagccggat aatctggagc ggatggtctc
     4201 gatcctctcg tttgttgcgg tcaggctgtt acagctcaga gaaagcttca cgccgccgca
     4261 agcactcagg gcgcaagggc tgctaaagga agcggaacac gtagaaagcc agtccgcaga
     4321 aacggtgctg accccggatg aatgtcagct actgggctat ctggacaagg gaaaacgcaa
     4381 gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg gcgatagcta gactgggcgg
     4441 ttttatggac agcaagcgaa ccggaattgc cagctggggc gccctctggg aaggttggga
     4501 agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat
     4561 caagatcctt taaggatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
     4621 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     4681 ttttgctgaa aggaggaact atatccggat atcccgcaag aggcccggca gtaccggcat
     4741 aaccaagcct atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt
     4801 gttagatttc atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca
     4861 ttaaagctta tcgatgataa gctgtcaaac atgagaattc ttgaagacga aagggcctcg
     4921 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     4981 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5041 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5101 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5161 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5221 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5281 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5341 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5401 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5461 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5521 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5581 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5641 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5701 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5761 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5821 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5881 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5941 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6001 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6061 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6121 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6181 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6241 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     6301 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6361 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6421 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6481 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6541 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6601 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6661 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6721 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6781 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     6841 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     6901 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     6961 tatatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     7021 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     7081 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     7141 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     7201 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     7261 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     7321 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     7381 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     7441 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     7501 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     7561 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     7621 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     7681 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     7741 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     7801 atgcgcaccc gtggccagga cccaacgctg cccgagatgc gccgcgtgcg gctgctggag
     7861 atggcggacg cgatggatat gttctgccaa gggttggttt gcgcattcac agttctccgc
     7921 aagaattgat tggctccaat tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt
     7981 ccattcaggt cgaggtggcc cggctccatg caccgcgacg caacgcgg
//
