LOCUS       sequence-436006-        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) T2C, C90A, T198C
FEATURES             Location/Qualifiers
     source          1..6682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             925..1116
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1110..1132)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1218..1357
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1273..1292
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1372..1389)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1543..2131)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1623..1642)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2302..3093,3094..3162))
                     /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     2925..2944
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3163..3267)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3293..3748)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3430..3451)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3642..3661
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(3785..3832)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3828..3846
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3899..3916)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4997..5017)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(5042..5203)
                     /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             complement(5219..5236)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             5249..5271
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    5286..5310
                     /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)."
     primer_bind     complement(5310..5329)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5311..5329)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5413..5432)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5638..5715
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5716..6682,1..116)
                     /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     complement(5735..5754)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg
       61 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
      121 aacgcaatta atgtaagtta gctcactcat taggcaccgg gatctcgacc gatgcccttg
      181 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca
      241 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc
      301 attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta
      361 ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc
      421 ggcgagaagc aggccattat cgccggcatg gcggccccac gggtgcgcat gattgtgctc
      481 ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca
      541 ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca
      601 acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc
      661 tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct
      721 acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc
      781 atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca
      841 gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga
      901 aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc
      961 cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat
     1021 gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc
     1081 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     1141 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     1201 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
     1261 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccattgc ggtgtgaaat
     1321 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
     1381 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1441 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1501 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1561 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1621 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1681 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1741 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1801 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1861 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1921 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1981 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2041 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2101 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2161 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2221 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2281 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2341 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2401 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     2461 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     2521 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     2581 gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc
     2641 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     2701 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     2761 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     2821 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     2881 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     2941 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3001 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3061 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3121 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3181 ttattgaagc atttatcagg gttattgtct cattagcgga tacatatttg aatgtattta
     3241 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta
     3301 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3361 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3421 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3481 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3541 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3601 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3661 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3721 gcgcttaatg cgccgctaca gggcgcgtcc cattcgccaa tccggatata gttcctcctt
     3781 tcagcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct agttattgct
     3841 cagcggtggc agcagccaac tcagcttcct ttcgggcttt gttagcagcc ggatctcagt
     3901 ggtggtggtg gtggtgctcg agtgcggccg caagcttgtc gacggagctc gaattcggat
     3961 cctattaatc tgacttgtgg gagtaaaggt ccttaagtgc ttttagctcc tcaatcaatg
     4021 ttttgttttg gttttcaagc actgccactc tgttctctaa agatttcaca tattctttct
     4081 tctttctacg agattctctt gctgcttccc tgttcttcat tagacgaacc tctctctttc
     4141 gtgcctatta ttcagcaggc tgtgtaggaa gtgctgggga ggacgccata acaactccag
     4201 gggcaatggt gctagtgggt gctgtgcgaa tctggtatgt ttgtacatca ccagaggcag
     4261 cttgaacaac aacttggttg ctgggcacta gaatctgctg tccatcagtg gtctgtgcat
     4321 attgtagaat ggtagtaccc ggctgagtgg cagctgcatt ggtcatggtt aatgtctgca
     4381 ggccctgtac cccatcgcaa ccattgttag ccagctgtat tgctcctccc tgggtaatgg
     4441 caatatactg cccactgcta gtttggtaaa tcggggttgg cactgttaca gtggtgatgg
     4501 caggggctga agtctcttct tctgattttt cttcttcaat ccttggcacc cctggtgcat
     4561 cagaagataa gtcattcaaa attttcctgt aggaaggcct ccttgaaagg atttcccttc
     4621 gtttttggga atcagttaca ctatccacag actcctgtga atcttcactt tctgcaatag
     4681 ttgaaatctg agttccggag aaaagtcttt ttaagtcctt agcggaagac tgaactgttt
     4741 ggacttgtgg agactgaata actgatggct gggccgcctg gatgacccca tggacctgga
     4801 ctgtctgccc attgggcagc tgcactaagg ttacagtggg agcagatgac gtcgcatgag
     4861 ctgctggcat ggatacctgg gctaatgtgg caatctgtgg ctgggcttga actgtcattt
     4921 gttgactttc agcttctgtt acagcagcat ctccactctg ctggttgtct gctccagagt
     4981 ccatgcacat atgggagccc tgaaaataca ggttttcgcc gctgctgccg ctgctgaatt
     5041 cttcagtaac ggtgaaggtc ttagtcgcat cgtcgtaagt ccattcaccg tcaacaccgt
     5101 tgtcgttagc gtactgtttg aaaacttttt ctgcagtcgc agcatcaaca gcttcggtag
     5161 tggtttcgcc tttcaaagtt ttaccgttaa ggatcagttt gtaagtcatg ccgctgctgt
     5221 gatgatgatg atgatggccc atggtatatc tccttcttaa agttaaacaa aattatttct
     5281 agaggggaat tgttatccgc tcacaattcc cctatagtga gtcgtattaa tttcgcggga
     5341 tcgagatctc gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg
     5401 cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg
     5461 ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt
     5521 tgggcgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc
     5581 tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga gatcccggac
     5641 accatcgaat ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa
     5701 ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc
     5761 tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg
     5821 gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa
     5881 ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg
     5941 ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg
     6001 gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc
     6061 gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct
     6121 gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc
     6181 atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc
     6241 gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt
     6301 ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa
     6361 cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag
     6421 ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc
     6481 gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat
     6541 accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg
     6601 ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc
     6661 aatcagctgt tgcccgtctc ac
//
