LOCUS       sequence-353458-        6177 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial coexpression of CBP TAZ1 with CITED2/Hif1a activation 
            domain constructs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6177)
  AUTHORS   Appling FD, Berlow RB, Stanfield RL, Dyson HJ, Wright PE
  TITLE     The molecular basis of allostery in a facilitated dissociation 
            process.
  JOURNAL   Structure. 2021 Sep 11. pii: S0969-2126(21)00261-6. doi: 
            10.1016/j.str.2021.07.011.
  PUBMED    34520739
REFERENCE   2  (bases 1 to 6177)
  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: "Structure. 
            2021 Sep 11. pii: S0969-2126(21)00261-6. doi: 
            10.1016/j.str.2021.07.011."
COMMENT     Sequence Label: HIsGB1-FusionL63A + CBP TAZ1(340-439)
FEATURES             Location/Qualifiers
     source          1..6177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             492..514
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             complement(736..753)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(778..939)
                     /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             complement(955..972)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             985..1007
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1022..1046
                     /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)."
     primer_bind     complement(1046..1065)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1047..1065)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1149..1168)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1374..1451
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1452..2534
                     /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     complement(1471..1490)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3343..3534
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3528..3550)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3636..3775
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3691..3710
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3790..3807)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3961..4549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4041..4060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4720..5511,5512..5580))
                     /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     5343..5362
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5581..5685)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5711..6166)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5848..5869)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6060..6079
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttac
      181 tatcgcttgt cactggcatt tttcaaaggg aggcaaacag gacagtcatg tcgtgtgcag
      241 ttcttccaat gagagatgat ttgtcgtgaa gatgcacaat gggcaacttg gcaggctttc
      301 ccagcctgac aatgtgtcat gtgattcaaa acgtttttca tggttcgaca gtgtgggaga
      361 gaacaggctc gaacctctcc atttgcttgc tctcgtctct gacatttgtg ggcatgaagc
      421 agtagaacca gctgctgctg tatcagtttg cgtttttcag ggtctgctgt ggggcctgtt
      481 gccatggtat atctccttct taaagttaaa caaaattatt tctagagggg atccctatca
      541 gttaacttga tccgcagctc tgagtaattc ttcaccctgc agtaggtttc tgctgccttg
      601 tataggagca ttaacttcac aatcataact ggtcagttct ggtaactcct tgatgcggtc
      661 caaacccatt tctatcacca aggacataag aacttcctcg tcgatgaaat cagtgtctat
      721 gacattggac atatgggaac cacgcggaac caggccgctg ctgccgctgc tgaattcttc
      781 agtaacggtg aaggtcttag tcgcatcgtc gtaagtccat tcaccgtcaa caccgttgtc
      841 gttagcgtac tgtttgaaaa ctttttctgc agtcgcagca tcaacagctt cggtagtggt
      901 ttcgcctttc aaagttttac cgttaaggat cagtttgtaa gtcatgccgc tgctgtgatg
      961 atgatgatga tggcccatcg tatatctcct tcttaaagtt aaacaaaatt atttctagag
     1021 gggaattgtt atccgctcac aattccccta tagtgagtcg tattaatttc gcgggatcga
     1081 gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt
     1141 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct
     1201 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg
     1261 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
     1321 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca
     1381 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
     1441 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
     1501 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
     1561 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
     1621 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
     1681 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
     1741 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
     1801 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
     1861 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
     1921 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
     1981 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
     2041 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
     2101 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
     2161 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
     2221 ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg
     2281 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
     2341 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
     2401 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
     2461 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     2521 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga
     2581 tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
     2641 atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg
     2701 ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc
     2761 ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt
     2821 cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccccacgg
     2881 gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact
     2941 ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg
     3001 tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa
     3061 acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg
     3121 ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt
     3181 tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac
     3241 cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc
     3301 attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa
     3361 accgccctta acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc
     3421 aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat
     3481 gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     3541 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     3601 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
     3661 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     3721 accattgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc
     3781 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3841 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3901 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3961 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4021 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4081 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4141 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4201 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4261 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4321 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4381 taactacggc tacactagaa ggacagtatt tggtatctgt gctctgctga agccagttac
     4441 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4501 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4561 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4621 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4681 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4741 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4801 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4861 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4921 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4981 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg
     5041 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5101 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5161 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5221 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5281 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5341 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5401 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5461 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5521 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5581 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5641 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5701 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     5761 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     5821 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     5881 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     5941 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     6001 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     6061 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     6121 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgcca
//
