LOCUS Exported 8028 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION This vector encode a His6-tagged proteinA-Tnp(hyperactive form) fusion protein.. ACCESSION . VERSION . KEYWORDS pET15bpATnp SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8028) AUTHORS Benjamin Carter, Wai Lim Ku, Qingsong Tang, Keji Zhao TITLE Mapping Histone Modifications in Low Cell Number and Single Cells Using Antibody-guided Chromatin Tagmentation (ACT-seq) JOURNAL BioRxiv, 2019 REFERENCE 2 (bases 1 to 8028) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8028 /organism="synthetic DNA construct" /mol_type="other DNA" 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(4860..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 5106..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 //