LOCUS       sequence-436969-        6475 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 6475)
  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 6475)
  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(1-280) T160C
FEATURES             Location/Qualifiers
     source          1..6475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             678..695
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(748..766)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      762..809
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      846..1301
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(933..952)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1143..1164
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1327..1431
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1432..1500,1501..2292)
                     /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(1650..1669)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2463..3051
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2952..2971
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3205..3222
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3237..3376
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3302..3321)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3462..3484
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3478..3669)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4478..5560)
                     /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     5522..5541
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5561..5638)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5844..5863
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5947..5966
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5947..5965
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5966..5990
                     /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             6005..6027
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             6040..6057
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6073..6234
                     /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             6259..6279
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 tccaggtcca tggggtcatc caggcggccc agccatcagt tattcagtct ccacaagtcc
       61 aaacagttca gtcttcctgt aaggacttaa aaagactttt ctccggaact cagatttcaa
      121 ctattgcaga aagtgaagat tcacaggagt ctgtggatag tgtaactgat tcccaaaaac
      181 gaagggaaat cctttcaagg aggccttcct acaggaaaat tttgaatgac ttatcttctg
      241 atgcaccagg ggtgccaagg attgaagaag aaaaatcaga agaagagtgt tcagcccctg
      301 ccatcaccac tgtaacagtg ccaaccccga tttaccaaac tagcagtggg cagtatattg
      361 ccattaccca gggaggagca atacagctgg ctaacaatgg taccgatggg gtacagggcc
      421 tgcagacatt aaccatgacc aatgcagctg ccactcagcc gggtactacc attctacaat
      481 atgcacagac cactgatgga cagcagattc tagtgcccag caaccaagtt gttgttcaag
      541 ctgcctctgg tgatgtacaa acataccaga ttcgcacagc acccactagc accattgccc
      601 ctggagttgt tatggcgtcc tccccagcac ttcctacaca gcctgctgaa taatagaagc
      661 ttgcggccgc actcgagcac caccaccacc accactgaga tccggctgct aacaaagccc
      721 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
      781 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga
      841 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
      901 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
      961 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     1021 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     1081 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     1141 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     1201 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     1261 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     1321 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1381 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     1441 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     1501 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     1561 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     1621 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     1681 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     1741 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     1801 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     1861 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     1921 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     1981 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2041 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2101 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2161 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2221 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     2281 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     2341 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     2401 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2461 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2521 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2581 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     2641 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2701 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     2761 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     2821 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     2881 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     2941 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3001 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3061 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3121 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3181 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     3241 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca
     3301 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
     3361 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3421 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3481 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     3541 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     3601 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     3661 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat
     3721 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
     3781 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
     3841 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca
     3901 tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact
     3961 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca
     4021 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca
     4081 accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc
     4141 cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac
     4201 gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt
     4261 cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc
     4321 tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc
     4381 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc
     4441 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4501 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4561 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct
     4621 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgt cccagcaggc
     4681 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt
     4741 cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca
     4801 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat
     4861 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg
     4921 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg
     4981 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca
     5041 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg
     5101 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa
     5161 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa
     5221 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca
     5281 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt
     5341 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt
     5401 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg
     5461 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac
     5521 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac
     5581 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg
     5641 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg
     5701 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
     5761 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     5821 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     5881 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg
     5941 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa
     6001 taattttgtt taactttaag aaggagatat accatgggcc atcatcatca tcatcacagc
     6061 agcggcatga cttacaaact gatccttaac ggtaaaactt tgaaaggcga aaccactacc
     6121 gaagctgttg atgctgcgac tgcagaaaaa gttttcaaac agtacgctaa cgacaacggt
     6181 gttgacggtg aatggactta cgacgatgcg actaagacct tcaccgttac tgaagaattc
     6241 agcagcggca gcagcggcga aaacctgtat tttcagggct cccatatgac catggactct
     6301 ggagcagaca accagcagag tggagatgct gctgtaacag aagctgaaag tcaacaaatg
     6361 acagttcaag cccagccaca gattgccaca ttagcccagg tatccatgcc agcagctcat
     6421 gcgacgtcat ctgctcccac tgtaacctta gtgcagctgc ccaatgggca gacag
//
