LOCUS       sequence-193349-        5833 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial coexpression of CBP NCBD and NCoA3 binding domains.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5833)
  AUTHORS   Demarest SJ, Martinez-Yamout M, Chung J, Chen H, Xu W, Dyson HJ, 
            Evans RM, Wright PE
  TITLE     Mutual synergistic folding in recruitment of CBP/p300 by p160 
            nuclear receptor coactivators.
  JOURNAL   Nature. 2002 Jan 31;415(6871):549-53.
  PUBMED    11823864
REFERENCE   2  (bases 1 to 5833)
  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: "Nature."; 
            date: "2002-01-31"; volume: "415(6871)"; pages: "549-53"
COMMENT     Sequence Label: pET-NCBD + ACTR
FEATURES             Location/Qualifiers
     source          1..5833
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      14..602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     503..522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     756..773
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    788..927
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(853..872)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1013..1035
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1029..1220)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2029..3111)
                     /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     3073..3092
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3112..3189)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3395..3414
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3498..3517
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3498..3516
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3517..3541
                     /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             3556..3578
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             3792..3814
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4062..4079
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4132..4150)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4146..4193
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4230..4685
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4317..4336)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4527..4548
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4711..4815
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4816..4884,4885..5676)
                     /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(5034..5053)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
       61 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      121 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
      181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      421 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      601 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      661 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      721 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      781 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg
      841 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
      901 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
      961 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     1021 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc
     1081 tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg
     1141 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg
     1201 gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta
     1261 atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc
     1321 cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga
     1381 aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt
     1441 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
     1501 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
     1561 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa
     1621 ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc
     1681 acccgtgggg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg
     1741 ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg
     1801 ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc
     1861 ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc
     1921 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     1981 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     2041 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     2101 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     2161 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     2221 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     2281 ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt
     2341 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     2401 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     2461 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     2521 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     2581 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     2641 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     2701 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     2761 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     2821 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     2881 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     2941 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     3001 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     3061 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     3121 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     3181 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     3241 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     3301 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     3361 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     3421 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     3481 tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc
     3541 ccctctagaa ataattttgt ttaactttaa gaaggagata tacatatgcc aaacaggagc
     3601 atctcgccaa gtgccctgca agacctgcta cggaccctaa agtcacccag ctctcctcag
     3661 cagcagcagc aggtgctgaa catccttaaa tcaaacccac agctaatggc agctttcatc
     3721 aaacagcgca cagccaagta tgtggccaat cagcctggca tgcagtgata aggatcccct
     3781 ctagaaataa ttttgtttaa ctttaagaag gagatatacc atgggcactc aaaataggcc
     3841 tcttcttagg aattccctgg atgatcttgt tgggccacct tccaacctgg aaggccagag
     3901 tgacgaaaga gcattattgg accagctgca cactcttctc agcaacacag atgccacagg
     3961 cctggaagaa attgacagag ctttgggcat tcctgaactt gtcaatcagg gacaggcatt
     4021 agagcccaaa caggattgat aagcttgcgg ccgcactcga gcaccaccac caccaccact
     4081 gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     4141 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     4201 gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     4261 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4321 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4381 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4441 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4501 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4561 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4621 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     4681 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4741 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4801 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4861 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4921 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4981 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5041 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5101 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5161 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5221 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5281 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5341 cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5401 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5461 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5521 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5581 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5641 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5701 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5761 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5821 cccgtagaaa aga
//
