LOCUS       sequence-192994-        5670 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of mouse CBP KIX domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5670)
  AUTHORS   Radhakrishnan I, Perez-Alvarado GC, Parker D, Dyson HJ, Montminy MR,
            Wright PE
  TITLE     Structural analyses of CREB-CBP transcriptional 
            activator-coactivator complexes by NMR spectroscopy: implications 
            for mapping the boundaries of structural domains.
  JOURNAL   J Mol Biol. 1999 Apr 16;287(5):859-65.
  PUBMED    10222196
REFERENCE   2  (bases 1 to 5670)
  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: "J Mol 
            Biol."; date: "1999-04-16"; volume: "287(5)"; pages: "859-65"
COMMENT     Sequence Label: pET-KIX (586-672)
FEATURES             Location/Qualifiers
     source          1..5670
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      30..618
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     519..538
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     772..789
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    804..943
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(869..888)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1029..1051
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1045..1236)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2045..3127)
                     /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     3089..3108
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3128..3205)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3411..3430
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3514..3533
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3514..3532
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3533..3557
                     /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             3572..3594
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3915..3932
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3985..4003)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3999..4046
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4083..4538
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4170..4189)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4380..4401
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4564..4668
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4669..4737,4738..5529)
                     /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(4887..4906)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
       61 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
      121 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
      181 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
      241 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
      301 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
      361 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
      421 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
      481 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
      541 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
      601 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
      661 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
      721 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
      781 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
      841 atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
      901 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
      961 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     1021 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     1081 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     1141 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     1201 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     1261 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     1321 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     1381 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     1441 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     1501 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
     1561 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     1621 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     1681 gagcacgatc atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc
     1741 gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa
     1801 taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat
     1861 gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag
     1921 tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct
     1981 caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt
     2041 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     2101 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca
     2161 gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc
     2221 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga
     2281 tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc
     2341 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca
     2401 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca
     2461 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt
     2521 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg
     2581 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat
     2641 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa
     2701 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga
     2761 tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc
     2821 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa
     2881 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca
     2941 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg
     3001 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca
     3061 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg
     3121 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa
     3181 aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc
     3241 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
     3301 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     3361 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     3421 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     3481 gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt
     3541 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatataca
     3601 tatgggtgtt cgaaaaggct ggcatgaaca tgtgactcag gacctacgga gtcatctagt
     3661 ccataaactc gttcaagcca tcttcccaac tccagaccct gcagctctga aagatcgccg
     3721 catggagaac ctggttgcct atgctaagaa agtggaggga gacatgtatg agtctgctaa
     3781 tagcagggat gaatactatc atttattagc agagaaaatc tataaaatac aaaaagaact
     3841 agaagaaaag cggaggtcac gtttatgagg atccgaattc gagctccgtc gacaagcttg
     3901 cggccgcact cgagcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa
     3961 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     4021 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg
     4081 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4141 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4201 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4261 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4321 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4381 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4441 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4501 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     4561 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4621 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4681 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4741 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4801 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4861 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4921 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4981 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5041 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5101 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5161 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     5221 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5281 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5341 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5401 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5461 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5521 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5581 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5641 taacgtgagt tttcgttcca ctgagcgtca
//
