LOCUS       sequence-436005-        6682 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 6682)
  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 6682)
  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) C90A, T172C, T259C
FEATURES             Location/Qualifiers
     source          1..6682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(166..1248)
                     /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     1210..1229
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1249..1326)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1532..1551
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1635..1654
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1635..1653
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1654..1678
                     /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             1693..1715
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1728..1745
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1761..1922
                     /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             1947..1967
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3048..3065
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3118..3136)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3132..3179
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3216..3671
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3303..3322)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3513..3534
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3697..3801
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3802..3870,3871..4662)
                     /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(4020..4039)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4833..5421
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5322..5341
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5575..5592
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5607..5746
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5672..5691)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5832..5854
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5848..6039)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
       61 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
      121 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
      181 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
      241 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
      301 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
      361 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
      421 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
      481 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
      541 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
      601 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
      661 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
      721 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
      781 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
      841 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
      901 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
      961 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     1021 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     1081 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     1141 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     1201 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     1261 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     1321 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     1381 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     1441 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     1501 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     1561 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct
     1621 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
     1681 tctagaaata attttgttta actttaagaa ggagatatac catgggccat catcatcatc
     1741 atcacagcag cggcatgact tacaaactga tccttaacgg taaaactttg aaaggcgaaa
     1801 ccactaccga agctgttgat gctgcgactg cagaaaaagt tttcaaacag tacgctaacg
     1861 acaacggtgt tgacggtgaa tggacttacg acgatgcgac taagaccttc accgttactg
     1921 aagaattcag cagcggcagc agcggcgaaa acctgtattt tcagggctcc catatgacca
     1981 tggactctgg agcagacaac cagcagagtg gagatgctgc tgtaacagaa gctgaaagtc
     2041 aacaaatgac agttcaagcc cagccacaga ttgccacatt agcccaggta tccatgccag
     2101 cagctcatgc gacgtcatct gctcccactg taaccttagt gcagctgccc aatgggcaga
     2161 cagtccaggt ccatggggtc atccaggcgg cccagccatc agttattcag tctccacaag
     2221 tccaaacagt tcagtcttcc gcgaaggact taaaaagact tttctccgga actcagattt
     2281 caactattgc agaaagtgaa gattcacagg agtctgtgga tagtgtaact gattcccaaa
     2341 aacgaaggga aatcctttca aggaggcctt cctacaggaa aattttgaat gacttatctt
     2401 ctgatgcacc aggggtgcca aggattgaag aagaaaaatc agaagaagag acttcagccc
     2461 ctgccatcac cactgtaaca gtgccatgcc cgatttacca aactagcagt gggcagtata
     2521 ttgccattac ccagggagga gcaatacagc tggctaacaa tggtaccgat ggggtacagg
     2581 gcctgcagac attaaccatg accaatgcag ctgccactca gccgggtact accattctac
     2641 aatatgcaca gaccactgat ggacagcaga ttctagtgcc cagcaaccaa gttgttgttc
     2701 aagctgcctc tggtgatgta caaacatacc agattcgcac agcaccctgc agcaccattg
     2761 cccctggagt tgttatggcg tcctccccag cacttcctac acagcctgct gaataatagg
     2821 cacgaaagag agaggttcgt ctaatgaaga acagggaagc agcaagagaa tctcgtagaa
     2881 agaagaaaga atatgtgaaa tctttagaga acagagtggc agtgcttgaa aaccaaaaca
     2941 aaacattgat tgaggagcta aaagcactta aggaccttta ctcccacaag tcagattaat
     3001 aggatccgaa ttcgagctcc gtcgacaagc ttgcggccgc actcgagcac caccaccacc
     3061 accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     3121 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     3181 tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag
     3241 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3301 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3361 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3421 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3481 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3541 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3601 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3661 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3721 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3781 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3841 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3901 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3961 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4021 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4081 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4141 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4201 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4261 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4321 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     4381 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4441 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4501 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4561 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4621 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4681 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4741 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4801 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4861 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4921 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     4981 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5041 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5101 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5161 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5221 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5281 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5341 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5401 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5461 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5521 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     5581 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     5641 ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5701 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     5761 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     5821 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     5881 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     5941 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     6001 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     6061 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     6121 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     6181 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     6241 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     6301 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     6361 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     6421 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     6481 caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc
     6541 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     6601 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     6661 aatgacccag agcgctgccg gc
//
