LOCUS       sequence-193363-        6141 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of mouse CBP TAZ1 domain as coexpression with 
            Hif1a TAD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6141)
  AUTHORS   Sugase K, Landes MA, Wright PE, Martinez-Yamout M
  TITLE     Overexpression of post-translationally modified peptides in 
            Escherichia coli by co-expression with modifying enzymes.
  JOURNAL   Protein Expr Purif. 2008 Feb;57(2):108-15. Epub 2007 Nov 1.
  PUBMED    18054500
REFERENCE   2  (bases 1 to 6141)
  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: "Protein 
            Expr Purif."; date: "2008-02"; volume: "57(2)"; pages: "108-15. Epub
            2007 Nov 1"
COMMENT     Sequence Label: HisGB1-Hif1a(776-826) + CBP TAZ1(340-439)
FEATURES             Location/Qualifiers
     source          1..6141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      21..609
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     510..529
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     763..780
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    795..934
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(860..879)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1020..1042
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1036..1227)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2036..3118)
                     /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     3080..3099
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3119..3196)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3402..3421
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3505..3524
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3505..3523
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3524..3548
                     /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             3563..3585
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3598..3615
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3631..3792
                     /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             3817..3834
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     RBS             4020..4042
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4377..4394
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4447..4465)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4461..4508
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4545..5000
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4632..4651)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4842..4863
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5026..5130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5131..5199,5200..5991)
                     /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(5349..5368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
       61 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
      121 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
      181 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
      241 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
      301 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
      361 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
      421 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
      481 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
      541 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
      601 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
      661 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
      721 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
      781 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
      841 cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
      901 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
      961 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     1021 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     1081 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     1141 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     1201 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
     1261 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     1321 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     1381 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
     1441 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
     1501 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
     1561 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
     1621 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
     1681 catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt
     1741 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag
     1801 cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag
     1861 cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac
     1921 gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     1981 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     2041 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2101 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     2161 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     2221 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     2281 agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg
     2341 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     2401 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     2461 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     2521 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     2581 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     2641 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     2701 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     2761 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     2821 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     2881 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     2941 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     3001 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     3061 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     3121 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     3181 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag
     3241 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
     3301 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     3361 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     3421 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     3481 atcgagatct cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata
     3541 acaattcccc tctagaaata attttgttta actttaagaa ggagatatac gatgggccat
     3601 catcatcatc atcacagcag cggcatgact tacaaactga tccttaacgg taaaactttg
     3661 aaaggcgaaa ccactaccga agctgttgat gctgcgactg cagaaaaagt tttcaaacag
     3721 tacgctaacg acaacggtgt tgacggtgaa tggacttacg acgatgcgac taagaccttc
     3781 accgttactg aagaattcag cagcggcagc agcggcctgg ttccgcgtgg ttcccatatg
     3841 tctgatttag catgtagact gctggggcaa tcaatggatg aaagtggatt accacagctg
     3901 accagttatg attgtgaagt taatgctcct atacaaggca gcagaaacct actgcagggt
     3961 gaagaattac tcagagcttt ggatcaagtt aactgatagg gatcccctct agaaataatt
     4021 ttgtttaact ttaagaagga gatataccat ggcaacaggc cccacagcag accctgaaaa
     4081 acgcaaactg atacagcagc agctggttct actgcttcat gcccacaaat gtcagagacg
     4141 agagcaagca aatggagagg ttcgagcctg ttctctccca cactgtcgaa ccatgaaaaa
     4201 cgttttgaat cacatgacac attgtcaggc tgggaaagcc tgccaagttg cccattgtgc
     4261 atcttcacga caaatcatct ctcattggaa gaactgcaca cgacatgact gtcctgtttg
     4321 cctccctttg aaaaatgcca gtgacaagcg atagtaagct tgcggccgca ctcgagcacc
     4381 accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     4441 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     4501 gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg
     4561 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4621 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4681 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4741 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4801 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4861 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4921 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4981 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5041 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5101 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5161 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5221 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5281 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5341 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5401 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5461 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5521 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5581 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5641 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     5701 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5761 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5821 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5881 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5941 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6001 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6061 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6121 cactgagcgt cagaccccgt a
//
