LOCUS       sequence-353461-        6154 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial coexpression of human CITED2 CTAD with CBP TAZ1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6154)
  AUTHORS   Berlow RB, Dyson HJ, Wright PE
  TITLE     Hypersensitive termination of the hypoxic response by a disordered 
            protein switch.
  JOURNAL   Nature. 2017 Mar 16;543(7645):447-451. doi: 10.1038/nature21705. 
            Epub 2017 Mar 8.
  PUBMED    28273070
REFERENCE   2  (bases 1 to 6154)
  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/nature21705"; journalName: "Nature"; date: "2017-03-16- 
            16"; volume: "543"; issue: "7645"; pages: "447-451"
COMMENT     Sequence Label: HisGB1-CITED2(216-269)+CBPTAZ1(340-439)
FEATURES             Location/Qualifiers
     source          1..6154
                     /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             494..516
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             complement(713..730)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(755..916)
                     /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(932..949)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             962..984
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    999..1023
                     /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(1023..1042)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1024..1042)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1126..1145)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1351..1428
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1429..2511
                     /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(1448..1467)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3320..3511
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3505..3527)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3613..3752
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3668..3687
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3767..3784)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3938..4526)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4018..4037)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4697..5488,5489..5557))
                     /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     5320..5339
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5558..5662)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5688..6143)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5825..5846)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6037..6056
                     /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 gcaagcttct
      181 attatcgctt gtcactggca tttttcaaag ggaggcaaac aggacagtca tgtcgtgtgc
      241 agttcttcca atgagagatg atttgtcgtg aagatgcaca atgggcaact tggcaggctt
      301 tcccagcctg acaatgtgtc atgtgattca aaacgttttt catggttcga cagtgtggga
      361 gagaacaggc tcgaacctct ccatttgctt gctctcgtct ctgacatttg tgggcatgaa
      421 gcagtagaac cagctgctgc tgtatcagtt tgcgtttttc agggtctgct gtggggcctg
      481 ttgccatggt atatctcctt cttaaagtta aacaaaatta tttctagagg ggatcctatt
      541 agctcactct gctgggctgc tgtttgcaca cgaagtccgt cataaaatca aactcgtttt
      601 gccccagcca gagttcgggc agctccttga tgcggtccaa acccatttct atcaccaagg
      661 acataagaac ttcctcgtcg atgaaatcag tgtctatgac attggacata tgggaaccac
      721 gcggaaccag gccgctgctg ccgctgctga attcttcagt aacggtgaag gtcttagtcg
      781 catcgtcgta agtccattca ccgtcaacac cgttgtcgtt agcgtactgt ttgaaaactt
      841 tttctgcagt cgcagcatca acagcttcgg tagtggtttc gcctttcaaa gttttaccgt
      901 taaggatcag tttgtaagtc atgccgctgc tgtgatgatg atgatgatgg cccatggtat
      961 atctccttct taaagttaaa caaaattatt tctagagggg aattgttatc cgctcacaat
     1021 tcccctatag tgagtcgtat taatttcgcg ggatcgagat ctcgatcctc tacgccggac
     1081 gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca
     1141 tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg
     1201 gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg catgcaccat
     1261 tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc ctaatgcagg
     1321 agtcgcataa gggagagcgt cgagatcccg gacaccatcg aatggcgcaa aacctttcgc
     1381 ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta
     1441 acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg
     1501 aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag
     1561 ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt
     1621 ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa
     1681 tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc
     1741 gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt
     1801 aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg
     1861 gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa
     1921 gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg
     1981 ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat
     2041 ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc
     2101 ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt
     2161 gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt
     2221 ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg
     2281 ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg
     2341 ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg
     2401 aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     2461 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     2521 aattaatgta agttagctca ctcattaggc accgggatct cgaccgatgc ccttgagagc
     2581 cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat
     2641 gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt
     2701 cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg
     2761 aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga
     2821 gaagcaggcc attatcgccg gcatggcggc cccacgggtg cgcatgatcg tgctcctgtc
     2881 gttgaggacc cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat
     2941 acgcgagcga acgtgaagcg actgctgctg caaaacgtct gcgacctgag caacaacatg
     3001 aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag cgccctgcac
     3061 cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc
     3121 tgtattaacg aagcgctggc attgaccctg agtgattttt ctctggtccc gccgcatcca
     3181 taccgccagt tgtttaccct cacaacgttc cagtaaccgg gcatgttcat catcagtaac
     3241 ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt acccccatga acagaaatcc
     3301 cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt
     3361 tatcagaagc cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca
     3421 ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca gctgcctcgc
     3481 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     3541 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     3601 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt
     3661 aactatgcgg catcagagca gattgtactg agagtgcacc attgcggtgt gaaataccgc
     3721 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
     3781 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3841 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3901 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3961 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4021 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4081 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4141 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4201 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4261 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4321 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     4381 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4441 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4501 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4561 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4621 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4681 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4741 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4801 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4861 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4921 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4981 ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt
     5041 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5101 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5161 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5221 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5281 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5341 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5401 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5461 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5521 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5581 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5641 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt
     5701 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     5761 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     5821 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     5881 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     5941 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     6001 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     6061 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     6121 taatgcgccg ctacagggcg cgtcccattc gcca
//
