LOCUS Exported 6998 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Bacterial expression plasmid encoding X. laevis H2B residues 1-114(K114A) as a chimeric fusion to GyrA intein with chitin binding domain at the C-terminus. H2B fragment can be derivatized using MESNa.. ACCESSION . VERSION . KEYWORDS pTXB1-xlH2B(1-114)_K114A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6998) AUTHORS Morgan MT, Haj-Yahya M, Ringel AE, Bandi P, Brik A, Wolberger C TITLE Structural basis for histone H2B deubiquitination by the SAGA DUB module. JOURNAL Science. 2016 Feb 12;351(6274):725-8. doi: 10.1126/science.aac5681. PUBMED 26912860 REFERENCE 2 (bases 1 to 6998) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6998 /organism="synthetic DNA construct" /mol_type="other DNA" RBS 1..23 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 374..967 /codon_start=1 /gene="Mycobacterium xenopi gyrA" /product="modified GyrA intein (Southworth et al., 1999)" /label=Mxe GyrA intein /note="N-terminal cleavage of this intein can be induced by thiols." /translation="CITGDALVALPEGESVRIADIVPGARPNSDNAIDLKVLDRHGNPV LADRLFHSGEHPVYTVRTVEGLRVTGTANHPLLCLVDVAGVPTLLWKLIDEIKPGDYAV IQRSAFSVDCAGFARGKPEFAPTTYTVGVPGLVRFLEAHHRDPDAQAIADELTDGRFYY AKVASVTDAGVQPVYSLRVDTADHAFITNGFVSHA" CDS 998..1153 /codon_start=1 /gene="Bacillus circulans chiA" /product="chitin binding domain from chitinase A1 (Watanabe et al., 1994)" /label=CBD /translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV PALWQLQ" primer_bind complement(1218..1236) /label=T7 Term /note="T7 terminator, reverse primer" terminator 1232..1279 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" promoter 1339..1443 /gene="bla" /label=AmpR promoter CDS 1444..2304 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(1662..1681) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin complement(2346..2859) /direction=LEFT /label=M13 ori /note="M13 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2541..2562) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2753..2772 /label=F1ori-R /note="F1 origin, reverse primer" rep_origin 2970..3558 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3459..3478 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3712..3729 /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(3751..3770) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3911..3933 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3927..4118) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4675..5757) /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 5719..5738 /label=LacI-R /note="LacI, reverse primer" promoter complement(5758..5835) /gene="lacI" /label=lacI promoter terminator 5988..6074 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 6171..6257 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 6354..6440 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 6537..6623 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 6720..6806 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 6941..6960 /label=T7 /note="T7 promoter, forward primer" promoter 6941..6959 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 6960..6984 /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)." ORIGIN 1 tttgtttaac tttaagaagg agatatacat aatggccaag tccgctccag ccccgaagaa 61 aggctccaag aaagcggtga ccaagactca gaagaaagac gggaaaaagc gcaggaagac 121 aaggaaggag agttatgcca tttacgtgta caaggtgctg aagcaggtgc accccgatac 181 cggcatctcg tccaaggcca tgagcatcat gaactccttt gtcaacgatg tgtttgagcg 241 catcgcaggg gaagcctccc gcctggctca ttacaacaag cgctccacca tcacctcccg 301 ggagatccag accgcggtcc gactgctgct gcctggggag ttggccaaac acgccgtgtc 361 cgagggcacc gcgtgcatca cgggagatgc actagttgcc ctacccgagg gcgagtcggt 421 acgcatcgcc gacatcgtgc cgggtgcgcg gcccaacagt gacaacgcca tcgacctgaa 481 agtccttgac cggcatggca atcccgtgct cgccgaccgg ctgttccact ccggcgagca 541 tccggtgtac acggtgcgta cggtcgaagg tctgcgtgtg acgggcaccg cgaaccaccc 601 gttgttgtgt ttggtcgacg tcgccggggt gccgaccctg ctgtggaagc tgatcgacga 661 aatcaagccg ggcgattacg cggtgattca acgcagcgca ttcagcgtcg actgtgcagg 721 ttttgcccgc gggaaacccg aatttgcgcc cacaacctac acagtcggcg tccctggact 781 ggtgcgtttc ttggaagcac accaccgaga cccggacgcc caagctatcg ccgacgagct 841 gaccgacggg cggttctact acgcgaaagt cgccagtgtc accgacgccg gcgtgcagcc 901 ggtgtatagc cttcgtgtcg acacggcaga ccacgcgttt atcacgaacg ggttcgtcag 961 ccacgctact ggcctcaccg gtctgaactc aggcctcacg acaaatcctg gtgtatccgc 1021 ttggcaggtc aacacagctt atactgcggg acaattggtc acatataacg gcaagacgta 1081 taaatgtttg cagccccaca cctccttggc aggatgggaa ccatccaacg ttcctgcctt 1141 gtggcagctt caatgactgc aggaagggga tccggctgct aacaaagccc gaaaggaagc 1201 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg 1261 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggataactac gtcaggtggc 1321 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat 1381 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag 1441 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt 1501 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt 1561 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc 1621 cccgaagaac gtttcccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta 1681 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac 1741 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa 1801 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg 1861 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc 1921 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg 1981 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta 2041 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg 2101 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg 2161 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc 2221 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt 2281 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt 2341 gatttacccc ggttgataat cagaaaagcc ccaaaaacag gaagattgta taagcaaata 2401 tttaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct 2461 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagcccg 2521 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact 2581 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac 2641 ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga 2701 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga 2761 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca 2821 ccacacccgc cgcgcttaat gcgccgctac agggcgcgta aaaggatcta ggtgaagatc 2881 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca 2941 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc 3001 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta 3061 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt 3121 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc 3181 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg 3241 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg 3301 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag 3361 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc 3421 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat 3481 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg 3541 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc 3601 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt 3661 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca 3721 gtgagcgagg aagctatggt gcactctcag tacaatctgc tctgatgccg catagttaag 3781 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca 3841 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct 3901 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg 3961 aggcagctgc ggtaaagctc atcagcgtgg tcgtgcagcg attcacagat gtctgcctgt 4021 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag 4081 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga 4141 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt 4201 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg 4261 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agccgaacgc cagcaagacg 4321 tagcccagcg cgtcggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg 4381 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc 4441 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc 4501 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg 4561 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc 4621 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg 4681 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 4741 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg 4801 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct 4861 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga 4921 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga 4981 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt 5041 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca 5101 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc 5161 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg 5221 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat 5281 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca 5341 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact 5401 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc 5461 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac 5521 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg 5581 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc 5641 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac 5701 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt 5761 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg 5821 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc 5881 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgccggc 5941 atgccgccct ttcgtcttca agaattaatt cccaattccc caggcatcaa ataaaacgaa 6001 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc 6061 tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt 6121 ggcgggcagg acgcccgcca taaactgcca ggaattaatt ccccaggcat caaataaaac 6181 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc 6241 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag 6301 ggtggcgggc aggacgcccg ccataaactg ccaggaatta attccccagg catcaaataa 6361 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg 6421 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg 6481 gagggtggcg ggcaggacgc ccgccataaa ctgccaggaa ttaattcccc aggcatcaaa 6541 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga 6601 acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc 6661 ccggagggtg gcgggcagga cgcccgccat aaactgccag gaattaattc cccaggcatc 6721 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg 6781 tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac 6841 ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggaattgg ggatcggaat 6901 taattcccgg tttaaaccgg ggatctcgat cccgcgaaat taatacgact cactataggg 6961 gaattgtgag cggataacaa ttcccctcta gaaataat //