LOCUS       sequence-436008-        5881 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 5881)
  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 5881)
  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(85-160)
FEATURES             Location/Qualifiers
     source          1..5881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     113..134
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        297..401
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(402..470,471..1262)
                     /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(620..639)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1433..2021
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1922..1941
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2175..2192
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2207..2346
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2272..2291)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2432..2454
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2448..2639)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3448..4530)
                     /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     4492..4511
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4531..4608)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4814..4833
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4917..4936
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4917..4935
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4936..4960
                     /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             4975..4997
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5010..5027
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5043..5204
                     /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             5229..5249
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             5529..5546
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5599..5617)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5613..5660
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      join(5697..5881,1..271)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5784..5803)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
       61 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
      121 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
      181 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
      241 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
      301 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
      361 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
      421 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
      481 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
      541 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
      601 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
      661 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
      721 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
      781 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
      841 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
      901 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
      961 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     1021 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     1081 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     1141 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     1201 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     1261 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     1321 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     1381 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     1441 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     1501 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     1561 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     1621 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     1681 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     1741 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     1801 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     1861 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     1921 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     1981 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2041 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     2101 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     2161 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     2221 ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg
     2281 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     2341 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     2401 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     2461 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
     2521 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
     2581 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     2641 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
     2701 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag
     2761 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
     2821 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
     2881 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
     2941 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
     3001 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
     3061 cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg
     3121 gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga
     3181 gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag
     3241 cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc
     3301 atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag
     3361 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     3421 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3481 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3541 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     3601 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     3661 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     3721 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     3781 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     3841 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     3901 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     3961 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     4021 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     4081 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     4141 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     4201 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     4261 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     4321 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     4381 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     4441 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     4501 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     4561 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     4621 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     4681 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     4741 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     4801 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     4861 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat
     4921 acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt
     4981 taactttaag aaggagatat accatgggcc atcatcatca tcatcacagc agcggcatga
     5041 cttacaaact gatccttaac ggtaaaactt tgaaaggcga aaccactacc gaagctgttg
     5101 atgctgcgac tgcagaaaaa gttttcaaac agtacgctaa cgacaacggt gttgacggtg
     5161 aatggactta cgacgatgcg actaagacct tcaccgttac tgaagaattc agcagcggca
     5221 gcagcggcga aaacctgtat tttcagggca cagttcagtc ttcctgcaag gacttaaaaa
     5281 gacttttctc cggaactcag atttcaacta ttgcagaaag tgaagattca caggagtctg
     5341 tggatagtgt aactgattcc caaaaacgaa gggaaatcct ttcaaggagg ccttcctaca
     5401 ggaaaatttt gaatgactta tcttctgatg caccaggggt gccaaggatt gaagaagaaa
     5461 aatcagaaga agagacttaa taggatccga attcgagctc cgtcgacaag cttgcggccg
     5521 cactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag
     5581 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     5641 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc
     5701 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5761 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5821 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5881 t
//
