LOCUS       sequence-193357-        6091 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of mouse CBP TAZ2 domain by coexpression with 
            HisGB1 tagged STAT1 TAD .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6091)
  AUTHORS   Wojciak JM, Martinez-Yamout MA, Dyson HJ, Wright PE
  TITLE     Structural basis for recruitment of CBP/p300 coactivators by STAT1 
            and STAT2 transactivation domains.
  JOURNAL   EMBO J. 2009 Apr 8;28(7):948-58. doi: 10.1038/emboj.2009.30. Epub 
            2009 Feb 12.
  PUBMED    19214187
REFERENCE   2  (bases 1 to 6091)
  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: 
            "10.1038/emboj.2009.30"; journalName: "EMBO J"; date: "2009-04-8- 
            8"; volume: "28"; issue: "7"; pages: "948-58"
COMMENT     Sequence Label: HisGb1-STAT1(710-750) + CBP TAZ2 (1764-1855)
FEATURES             Location/Qualifiers
     source          1..6091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      30..618
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     519..538
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     772..789
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    804..943
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(869..888)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1029..1051
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1045..1236)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2045..3127)
                     /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     3089..3108
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3128..3205)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3411..3430
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3514..3533
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3514..3532
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3533..3557
                     /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             3572..3594
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3607..3624
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3640..3801
                     /codon_start=1
                     /product="B1 domain of Streptococcal protein G   "
                     /label=GB1
                     /note="effective as a solubilizing fusion partner (Cheng 
                     and Patel, 2004)"
                     /translation="YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDD
                     ATKTFTVTE"
     CDS             3826..3843
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     RBS             3998..4020
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4336..4353
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4406..4424)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4420..4467
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4504..4959
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4591..4610)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4801..4822
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4985..5089
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5090..5158,5159..5950)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5308..5327)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
       61 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
      121 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
      181 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
      241 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
      301 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
      361 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
      421 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
      481 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
      541 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
      601 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
      661 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
      721 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
      781 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
      841 atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
      901 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
      961 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     1021 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     1081 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     1141 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     1201 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     1261 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     1321 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     1381 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     1441 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     1501 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
     1561 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     1621 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     1681 gagcacgatc atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc
     1741 gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa
     1801 taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat
     1861 gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag
     1921 tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct
     1981 caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt
     2041 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     2101 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca
     2161 gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc
     2221 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga
     2281 tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc
     2341 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca
     2401 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca
     2461 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt
     2521 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg
     2581 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat
     2641 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa
     2701 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga
     2761 tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc
     2821 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa
     2881 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca
     2941 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg
     3001 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca
     3061 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg
     3121 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa
     3181 aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc
     3241 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
     3301 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     3361 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     3421 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     3481 gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt
     3541 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc
     3601 atgggccatc atcatcatca tcacagcagc ggcatgactt acaaactgat ccttaacggt
     3661 aaaactttga aaggcgaaac cactaccgaa gctgttgatg ctgcgactgc agaaaaagtt
     3721 ttcaaacagt acgctaacga caacggtgtt gacggtgaat ggacttacga cgatgcgact
     3781 aagaccttca ccgttactga agaattcagc agcggcagca gcggcctggt tccgcgtggt
     3841 tcccatatgt ctgaagttca cccttctaga cttcagacca cagacaacct gctccccatg
     3901 tctcctgagg agtttgacga ggtgtctcgg atagtgggct ctgtagaatt cgacagtatg
     3961 atgaacacag tataatagga tcccctctag aaataatttt gtttaacttt aagaaggaga
     4021 tataccatgg gaagcccaca ggaatcccgg cgtctcagca tccagcgctg catccagtcc
     4081 ctggtgcatg cctgccagtg tcgcaatgcc aactgctcac tgccgtcttg ccagaagatg
     4141 aagcgagtcg tgcagcacac caagggctgc aagcgcaaga ctaatggagg atgcccagtg
     4201 tgcaagcagc tcattgctct ttgctgctac cacgccaaac actgccaaga aaataaatgc
     4261 cctgtgccct tctgcctcaa catcaaacat aagctccgcc agcagcagta atagaagctt
     4321 gcggccgcac tcgagcacca ccaccaccac cactgagatc cggctgctaa caaagcccga
     4381 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     4441 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat
     4501 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4561 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4621 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4681 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4741 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4801 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4861 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4921 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     4981 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5041 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     5101 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5161 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     5221 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     5281 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5341 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5401 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5461 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5521 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5581 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat
     5641 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5701 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5761 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5821 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5881 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5941 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     6001 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6061 ttaacgtgag ttttcgttcc actgagcgtc a
//
