LOCUS       sequence-436007-        5935 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of CREB1 domains.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5935)
  AUTHORS   Martinez-Yamout MA, Nasir I, Shnitkind S, Ellis JP, Berlow RB, Kroon
            G, Deniz AA, Dyson HJ, Wright PE
  TITLE     Glutamine-rich regions of the disordered CREB transactivation domain
            mediate dynamic intra- and intermolecular interactions.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Nov 21;120(47):e2313835120. doi: 
            10.1073/pnas.2313835120. Epub 2023 Nov 14.
  PUBMED    37971402
REFERENCE   2  (bases 1 to 5935)
  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: "2023-11-21"; volume: "120(47):e2313835120. 
            doi"; pages: " 10.1073/pnas.2313835120. Epub 2023 Nov 14"
COMMENT     Sequence Label: H6Gb1-CREB(1-100)
FEATURES             Location/Qualifiers
     source          1..5935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(55..72)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(406..426)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(451..612)
                     /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(628..645)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             658..680
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    695..719
                     /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(719..738)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(720..738)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(822..841)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1047..1124
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1125..2207
                     /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(1144..1163)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3016..3207
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3201..3223)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3309..3448
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3364..3383
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3463..3480)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3634..4222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3714..3733)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4393..5184,5185..5253))
                     /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     5016..5035
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5254..5358)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5384..5839)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5521..5542)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5733..5752
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(5876..5923)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     join(5919..5935,1..2)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 ggtggcagca gccaactcag cttcctttcg ggctttgtta gcagccggat ctcagtggtg
       61 gtggtggtgg tgctcgagtg cggccgcaag cttctattaa gttccggaga aaagtctttt
      121 taagtcctta caggaagact gaactgtttg gacttgtgga gactgaataa ctgatggctg
      181 ggccgcctgg atgaccccat ggacctggac tgtctgccca ttgggcagct gcactaaggt
      241 tacagtggga gcagatgacg tcgcatgagc tgctggcatg gatacctggg ctaatgtggc
      301 aatctgtggc tgggcttgaa ctgtcatttg ttgactttca gcttctgtta cagcagcatc
      361 tccactctgc tggttgtctg ctccagagtc catggtcata tgggagccct gaaaatacag
      421 gttttcgccg ctgctgccgc tgctgaattc ttcagtaacg gtgaaggtct tagtcgcatc
      481 gtcgtaagtc cattcaccgt caacaccgtt gtcgttagcg tactgtttga aaactttttc
      541 tgcagtcgca gcatcaacag cttcggtagt ggtttcgcct ttcaaagttt taccgttaag
      601 gatcagtttg taagtcatgc cgctgctgtg atgatgatga tgatggccca tggtatatct
      661 ccttcttaaa gttaaacaaa attatttcta gaggggaatt gttatccgct cacaattccc
      721 ctatagtgag tcgtattaat ttcgcgggat cgagatctcg atcctctacg ccggacgcat
      781 cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac
      841 cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat
      901 ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct
      961 tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc
     1021 gcataaggga gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc tttcgcggta
     1081 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
     1141 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     1201 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     1261 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     1321 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     1381 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     1441 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     1501 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
     1561 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
     1621 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
     1681 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
     1741 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
     1801 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
     1861 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
     1921 cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
     1981 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     2041 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
     2101 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     2161 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     2221 aatgtaagtt agctcactca ttaggcaccg ggatctcgac cgatgccctt gagagccttc
     2281 aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact
     2341 gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt cattttcggc
     2401 gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt attcggaatc
     2461 ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag
     2521 caggccatta tcgccggcat ggcggcccca cgggtgcgca tgatcgtgct cctgtcgttg
     2581 aggacccggc taggctggcg gggttgcctt actggttagc agaatgaatc accgatacgc
     2641 gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg
     2701 gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc ctgcaccatt
     2761 atgttccgga tctgcatcgc aggatgctgc tggctaccct gtggaacacc tacatctgta
     2821 ttaacgaagc gctggcattg accctgagtg atttttctct ggtcccgccg catccatacc
     2881 gccagttgtt taccctcaca acgttccagt aaccgggcat gttcatcatc agtaacccgt
     2941 atcgtgagca tcctctctcg tttcatcggt atcattaccc ccatgaacag aaatccccct
     3001 tacacggagg catcagtgac caaacaggaa aaaaccgccc ttaacatggc ccgctttatc
     3061 agaagccaga cattaacgct tctggagaaa ctcaacgagc tggacgcgga tgaacaggca
     3121 gacatctgtg aatcgcttca cgaccacgct gatgagcttt accgcagctg cctcgcgcgt
     3181 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     3241 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     3301 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact
     3361 atgcggcatc agagcagatt gtactgagag tgcaccattg cggtgtgaaa taccgcacag
     3421 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct
     3481 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3541 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3601 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3661 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3721 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3781 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3841 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3901 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3961 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4021 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     4081 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4141 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4201 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4261 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4321 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4381 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4441 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4501 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4561 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4621 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4681 tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg
     4741 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4801 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4861 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4921 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4981 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5041 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5101 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5161 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5221 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5281 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5341 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
     5401 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     5461 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     5521 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     5581 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     5641 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     5701 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     5761 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     5821 gcgccgctac agggcgcgtc ccattcgcca atccggatat agttcctcct ttcagcaaaa
     5881 aacccctcaa gacccgttta gaggccccaa ggggttatgc tagttattgc tcagc
//
