LOCUS       sequence-378016-        6073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Coexpression of CBP TAZ1 and CITED in E coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6073)
  AUTHORS   Berlow RB, Dyson HJ, Wright PE
  TITLE     Multivalency enables unidirectional switch-like competition between 
            intrinsically disordered proteins.
  JOURNAL   Proc Natl Acad Sci U S A. 2022 Jan 18;119(3). pii: 2117338119. doi: 
            10.1073/pnas.2117338119.
  PUBMED    35012986
REFERENCE   2  (bases 1 to 6073)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2022-01-18"; volume: "119(3). pii: 
            2117338119. doi"; pages: " 10.1073/pnas.2117338119"
COMMENT     Sequence Label: HisGB1-CITED(216-242)+CBPTAZ1(340-439)
FEATURES             Location/Qualifiers
     source          1..6073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..140
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(66..85)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     226..248
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(242..433)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1242..2324)
                     /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     2286..2305
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2325..2402)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2608..2627
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2711..2730
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2711..2729
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2730..2754
                     /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             2769..2791
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2804..2821
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2837..2998
                     /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             3023..3040
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     RBS             3156..3178
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3515..3532
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3585..3603)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3599..3646
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3683..4138
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3770..3789)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3980..4001
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4164..4268
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4269..4337,4338..5129)
                     /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(4487..4506)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5300..5888
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5789..5808
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6042..6059
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
       61 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
      121 gtgactgggt cattgctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
      181 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
      241 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
      301 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
      361 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
      421 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
      481 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
      541 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
      601 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
      661 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
      721 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
      781 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
      841 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
      901 gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
      961 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     1021 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     1081 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     1141 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     1201 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     1261 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     1321 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     1381 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     1441 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     1501 tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg
     1561 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     1621 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     1681 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     1741 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     1801 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     1861 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     1921 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     1981 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     2041 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     2101 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     2161 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     2221 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     2281 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     2341 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     2401 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
     2461 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa
     2521 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     2581 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     2641 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat
     2701 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta
     2761 gaaataattt tgtttaactt taagaaggag atataccatg ggccatcatc atcatcatca
     2821 cagcagcggc atgacttaca aactgatcct taacggtaaa actttgaaag gcgaaaccac
     2881 taccgaagct gttgatgctg cgactgcaga aaaagttttc aaacagtacg ctaacgacaa
     2941 cggtgttgac ggtgaatgga cttacgacga tgcgactaag accttcaccg ttactgaaga
     3001 attcagcagc ggcagcagcg gcctggttcc gcgtggttcc catatgtcca atgtcataga
     3061 cactgatttc atcgacgagg aagttcttat gtccttggtg atagaaatgg gtttggaccg
     3121 catcaaggag taataggatc ccctctagaa ataattttgt ttaactttaa gaaggagata
     3181 taccatggca acaggcccca cagcagaccc tgaaaaacgc aaactgatac agcagcagct
     3241 ggttctactg cttcatgccc acaaatgtca gagacgagag caagcaaatg gagaggttcg
     3301 agcctgttct ctcccacact gtcgaaccat gaaaaacgtt ttgaatcaca tgacacattg
     3361 tcaggctggg aaagcctgcc aagttgccca ttgtgcatct tcacgacaaa tcatctctca
     3421 ttggaagaac tgcacacgac atgactgtcc tgtttgcctc cctttgaaaa atgccagtga
     3481 caagcgataa tagaagcttg cggccgcact cgagcaccac caccaccacc actgagatcc
     3541 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     3601 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     3661 tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3721 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3781 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3841 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3901 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3961 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4021 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4081 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     4141 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4201 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4261 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4321 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4381 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4441 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4501 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4561 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4621 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4681 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4741 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4801 ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4861 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4921 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4981 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5041 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5101 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     5161 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5221 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5281 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5341 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5401 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     5461 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5521 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5581 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5641 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5701 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5761 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5821 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5881 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5941 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     6001 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     6061 aagcggaaga gcg
//
