LOCUS       sequence-378015-        6154 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Coexpression of CBP TAZ1 and CITED2 in E coli.
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     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 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"; 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: H6GB1-CITED2(216-269APAA) + CBP TAZ1(340-439)
FEATURES             Location/Qualifiers
     source          1..6154
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(69..1151)
                     /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     1113..1132
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1152..1229)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1435..1454
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1538..1557
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1538..1556
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1557..1581
                     /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             1596..1618
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1631..1648
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1664..1825
                     /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             1850..1867
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     RBS             2064..2086
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2423..2440
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2493..2511)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2507..2554
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2591..3046
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2678..2697)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2888..2909
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3072..3176
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3177..3245,3246..4037)
                     /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(3395..3414)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4208..4796
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4697..4716
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4950..4967
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4982..5121
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5047..5066)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5207..5229
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5223..5414)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg
       61 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
      121 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
      181 accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc
      241 aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc
      301 gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca
      361 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
      421 accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg
      481 gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga
      541 tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac
      601 agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct
      661 tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc
      721 ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta
      781 atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg
      841 acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat
      901 ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca
      961 atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc
     1021 tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc
     1081 accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt
     1141 actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg
     1201 cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc
     1261 ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa
     1321 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     1381 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     1441 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     1501 tccggcgtag aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa
     1561 ttgtgagcgg ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata
     1621 taccatgggc catcatcatc atcatcacag cagcggcatg acttacaaac tgatccttaa
     1681 cggtaaaact ttgaaaggcg aaaccactac cgaagctgtt gatgctgcga ctgcagaaaa
     1741 agttttcaaa cagtacgcta acgacaacgg tgttgacggt gaatggactt acgacgatgc
     1801 gactaagacc ttcaccgtta ctgaagaatt cagcagcggc agcagcggcc tggttccgcg
     1861 tggttcccat atgtccaatg tcatagacac tgatttcatc gacgaggaag ttcttatgtc
     1921 cttggtgata gaaatgggtt tggaccgcat caaggaggcg cccgcggcgt ggctggggca
     1981 aaacgagttt gattttatga cggacttcgt gtgcaaacag cagcccagca gagtgagcta
     2041 ataggatccc ctctagaaat aattttgttt aactttaaga aggagatata ccatggcaac
     2101 aggccccaca gcagaccctg aaaaacgcaa actgatacag cagcagctgg ttctactgct
     2161 tcatgcccac aaatgtcaga gacgagagca agcaaatgga gaggttcgag cctgttctct
     2221 cccacactgt cgaaccatga aaaacgtttt gaatcacatg acacattgtc aggctgggaa
     2281 agcctgccaa gttgcccatt gtgcatcttc acgacaaatc atctctcatt ggaagaactg
     2341 cacacgacat gactgtcctg tttgcctccc tttgaaaaat gccagtgaca agcgataata
     2401 gaagcttgcg gccgcactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa
     2461 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     2521 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt
     2581 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2641 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2701 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2761 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2821 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2881 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2941 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3001 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     3061 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3121 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3181 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3241 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3301 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3361 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3421 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3481 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3541 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3601 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3661 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3721 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3781 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3841 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3901 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3961 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4021 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4081 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4141 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4201 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4261 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4321 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     4381 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4441 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4501 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4561 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4621 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4681 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4741 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4801 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4861 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4921 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4981 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac
     5041 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
     5101 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     5161 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     5221 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
     5281 gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
     5341 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
     5401 tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata
     5461 ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta
     5521 ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc
     5581 actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag
     5641 cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc
     5701 agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt
     5761 ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta
     5821 aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt
     5881 ggggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca
     5941 gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc
     6001 atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc
     6061 tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc
     6121 cgcgcccacc ggaaggagct gactgggttg aagg
//
