LOCUS       sequence-443307-        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) C90A, T172C
FEATURES             Location/Qualifiers
     source          1..6475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(362..1444)
                     /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     1406..1425
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1445..1522)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1728..1747
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1831..1850
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1831..1849
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1850..1874
                     /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             1889..1911
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1924..1941
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1957..2118
                     /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             2143..2163
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3037..3054
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3107..3125)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3121..3168
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3205..3660
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3292..3311)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3502..3523
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3686..3790
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3791..3859,3860..4651)
                     /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(4009..4028)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4822..5410
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5311..5330
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5564..5581
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5596..5735
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5661..5680)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5821..5843
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5837..6028)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cacgatcatg cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa
       61 acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac
      121 cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac
      181 ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc
      241 ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
      301 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
      361 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
      421 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
      481 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
      541 ccacgctggt ttgtcccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
      601 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca
      661 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
      721 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
      781 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
      841 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
      901 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
      961 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     1021 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     1081 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     1141 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     1201 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     1261 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     1321 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     1381 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     1441 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     1501 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
     1561 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
     1621 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     1681 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
     1741 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     1801 tagaggatcg agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag
     1861 cggataacaa ttcccctcta gaaataattt tgtttaactt taagaaggag atataccatg
     1921 ggccatcatc atcatcatca cagcagcggc atgacttaca aactgatcct taacggtaaa
     1981 actttgaaag gcgaaaccac taccgaagct gttgatgctg cgactgcaga aaaagttttc
     2041 aaacagtacg ctaacgacaa cggtgttgac ggtgaatgga cttacgacga tgcgactaag
     2101 accttcaccg ttactgaaga attcagcagc ggcagcagcg gcgaaaacct gtattttcag
     2161 ggctcccata tgaccatgga ctctggagca gacaaccagc agagtggaga tgctgctgta
     2221 acagaagctg aaagtcaaca aatgacagtt caagcccagc cacagattgc cacattagcc
     2281 caggtatcca tgccagcagc tcatgcgacg tcatctgctc ccactgtaac cttagtgcag
     2341 ctgcccaatg ggcagacagt ccaggtccat ggggtcatcc aggcggccca gccatcagtt
     2401 attcagtctc cacaagtcca aacagttcag tcttccgcga aggacttaaa aagacttttc
     2461 tccggaactc agatttcaac tattgcagaa agtgaagatt cacaggagtc tgtggatagt
     2521 gtaactgatt cccaaaaacg aagggaaatc ctttcaagga ggccttccta caggaaaatt
     2581 ttgaatgact tatcttctga tgcaccaggg gtgccaagga ttgaagaaga aaaatcagaa
     2641 gaagagactt cagcccctgc catcaccact gtaacagtgc catgcccgat ttaccaaact
     2701 agcagtgggc agtatattgc cattacccag ggaggagcaa tacagctggc taacaatggt
     2761 accgatgggg tacagggcct gcagacatta accatgacca atgcagctgc cactcagccg
     2821 ggtactacca ttctacaata tgcacagacc actgatggac agcagattct agtgcccagc
     2881 aaccaagttg ttgttcaagc tgcctctggt gatgtacaaa cataccagat tcgcacagca
     2941 cccactagca ccattgcccc tggagttgtt atggcgtcct ccccagcact tcctacacag
     3001 cctgctgaat aatagaagct tgcggccgca ctcgagcacc accaccacca ccactgagat
     3061 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     3121 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     3181 actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3241 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3301 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3361 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3421 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3481 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3541 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3601 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     3661 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3721 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3781 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3841 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3901 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3961 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4021 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4081 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4141 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4201 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4261 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4321 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4381 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4441 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4501 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4561 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4621 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4681 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4741 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4801 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4861 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4921 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     4981 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5041 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5101 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5161 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5221 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5281 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5341 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5401 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5461 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     5521 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     5581 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     5641 ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     5701 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     5761 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     5821 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     5881 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     5941 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     6001 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
     6061 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg
     6121 aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg
     6181 accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc
     6241 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg
     6301 acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc
     6361 aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat
     6421 tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggag
//
