LOCUS       sequence-436000-        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)
FEATURES             Location/Qualifiers
     source          1..6475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     857..876
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(896..973)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1179..1198
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1282..1301
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1282..1300
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1301..1325
                     /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             1340..1362
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1375..1392
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1408..1569
                     /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             1594..1614
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2488..2505
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2558..2576)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2572..2619
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2656..3111
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2743..2762)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2953..2974
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3137..3241
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3242..3310,3311..4102)
                     /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(3460..3479)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4273..4861
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4762..4781
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5015..5032
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5047..5186
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5112..5131)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5272..5294
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5288..5479)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6288..6475,1..895))
                     /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"
ORIGIN
        1 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
       61 tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact
      121 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
      181 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
      241 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
      301 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
      361 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
      421 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
      481 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
      541 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
      601 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
      661 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
      721 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
      781 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
      841 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
      901 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
      961 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag
     1021 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     1081 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     1141 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     1201 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
     1261 gagatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca
     1321 attcccctct agaaataatt ttgtttaact ttaagaagga gatataccat gggccatcat
     1381 catcatcatc acagcagcgg catgacttac aaactgatcc ttaacggtaa aactttgaaa
     1441 ggcgaaacca ctaccgaagc tgttgatgct gcgactgcag aaaaagtttt caaacagtac
     1501 gctaacgaca acggtgttga cggtgaatgg acttacgacg atgcgactaa gaccttcacc
     1561 gttactgaag aattcagcag cggcagcagc ggcgaaaacc tgtattttca gggctcccat
     1621 atgaccatgg actctggagc agacaaccag cagagtggag atgctgctgt aacagaagct
     1681 gaaagtcaac aaatgacagt tcaagcccag ccacagattg ccacattagc ccaggtatcc
     1741 atgccagcag ctcatgcgac gtcatctgct cccactgtaa ccttagtgca gctgcccaat
     1801 gggcagacag tccaggtcca tggggtcatc caggcggccc agccatcagt tattcagtct
     1861 ccacaagtcc aaacagttca gtcttcctgt aaggacttaa aaagactttt ctccggaact
     1921 cagatttcaa ctattgcaga aagtgaagat tcacaggagt ctgtggatag tgtaactgat
     1981 tcccaaaaac gaagggaaat cctttcaagg aggccttcct acaggaaaat tttgaatgac
     2041 ttatcttctg atgcaccagg ggtgccaagg attgaagaag aaaaatcaga agaagagact
     2101 tcagcccctg ccatcaccac tgtaacagtg ccaaccccga tttaccaaac tagcagtggg
     2161 cagtatattg ccattaccca gggaggagca atacagctgg ctaacaatgg taccgatggg
     2221 gtacagggcc tgcagacatt aaccatgacc aatgcagctg ccactcagcc gggtactacc
     2281 attctacaat atgcacagac cactgatgga cagcagattc tagtgcccag caaccaagtt
     2341 gttgttcaag ctgcctctgg tgatgtacaa acataccaga ttcgcacagc acccactagc
     2401 accattgccc ctggagttgt tatggcgtcc tccccagcac ttcctacaca gcctgctgaa
     2461 taatagaagc ttgcggccgc actcgagcac caccaccacc accactgaga tccggctgct
     2521 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     2581 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     2641 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2701 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2761 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2821 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     2881 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2941 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3001 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3061 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     3121 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3181 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3241 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     3301 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     3361 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     3421 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     3481 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     3541 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     3601 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     3661 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     3721 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3781 gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3841 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3901 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3961 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4021 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     4081 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     4141 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     4201 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4261 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4321 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4381 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     4441 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4501 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4561 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4621 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4681 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4741 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4801 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4861 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4921 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     4981 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     5041 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg
     5101 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     5161 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     5221 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     5281 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     5341 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     5401 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     5461 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     5521 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     5581 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     5641 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     5701 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     5761 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     5821 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     5881 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     5941 cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     6001 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     6061 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     6121 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     6181 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
     6241 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     6301 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6361 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     6421 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctgg
//
