LOCUS       sequence-226718-        6998 bp    DNA     circular SYN 15-DEC-2025
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    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2016-02-12- 12"; volume: "351"; 
            issue: "6274"; pages: "725-8"
COMMENT     Sequence Label: pTXB1-xlH2B(1-114)_K114A
FEATURES             Location/Qualifiers
     source          1..6998
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             join(1444..1512,1513..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
//
