LOCUS       sequence-66239-e        6206 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GAL4 fusion to CREB-S133A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6206)
  AUTHORS   Sun P, Enslen H, Myung PS, Maurer RA
  TITLE     Differential activation of CREB by Ca2+/calmodulin-dependent protein
            kinases type II and type IV involves phosphorylation of a site that 
            negatively regulates activity.
  JOURNAL   Genes Dev. 1994 Nov 1;8(21):2527-39.
  PUBMED    7958915
REFERENCE   2  (bases 1 to 6206)
  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: "Genes 
            Dev."; date: "1994-11-1"; volume: "8(21)"; pages: "2527-39"
COMMENT     Sequence Label: pcDNAI-GAL4-CREB-S133A
FEATURES             Location/Qualifiers
     source          1..6206
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             11..451
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     primer_bind     complement(1489..1505)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1489..1505)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(1515..1533)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1516..1533)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     intron          1617..1682
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             1812..1832
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2104..2238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2141..2160)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2195..2214
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3084..3219
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3714..3733
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      complement(3846..4304)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3983..4004)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4198..4217
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4429..4447)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4515..4619
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4620..4688,4689..5480)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4838..4857)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     enhancer        5553..5932
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5933..6136
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6087..6107
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6181..6200
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6181..6199
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aagcttnnnn atgaagctac tgtcttctat cgaacaagca tgcgatattt gccgacttaa
       61 aaagctcaag tgctccaaag aaaaaccgaa gtgcgccaag tgtctgaaga acaactggga
      121 gtgtcgctac tctcccaaaa ccaaaaggtc tccgctgact agggcacatc tgacagaagt
      181 ggaatcaagg ctagaaagac tggaacagct atttctactg atttttcctc gagaagacct
      241 tgacatgatt ttgaaaatgg attctttaca ggatataaaa gcattgttaa caggattatt
      301 tgtacaagat aatgtgaata aagatgccgt cacagataga ttggcttcag tggagactga
      361 tatgcctcta acattgagac agcatagaat aagtgcgaca tcatcatcgg aagagagtag
      421 taacaaaggt caaagacagt tgactgtatc gccggaattc ggatccgact ctggagcaga
      481 caaccagcag agtggagatg ctgctgtaac agaagctgaa agtcaacaaa tgacagttca
      541 agcccagcca cagattgcca cattagccca ggtatccatg ccagcagctc atgcgacgtc
      601 atctgctccc actgtaacct tagtgcagct gcccaatggg cagacagtcc aggtccatgg
      661 ggtcatccag gcggcccagc catcagttat tcagtctcca caagtccaaa cagttcagtc
      721 ttcctgtaag gacttaaaaa gacttttctc cggaactcag atttcaacta ttgcagaaag
      781 tgaagattca caggagtctg tggatagtgt aactgattcc caaaaacgaa gggaaatcct
      841 ttcaaggagg cctgcctaca ggaaaatttt gaatgactta tcttctgatg caccaggggt
      901 gccaaggatt gaagaagaaa aatcagaaga agagacttca gcccctgcca tcaccactgt
      961 aacagtgcca accccgattt accaaactag cagtgggcag tatattgcca ttacccaggg
     1021 aggagcaata cagctggcta acaatggtac cgatggggta cagggcctgc agacattaac
     1081 catgaccaat gcagctgcca ctcagccggg tactaccatt ctacaatatg cacagaccac
     1141 tgatggacag cagattctag tgcccagcaa ccaagttgtt gttcaagctg cctctggtga
     1201 tgtacaaaca taccagattc gcacagcacc cactagcacc attgcccctg gagttgttat
     1261 ggcgtcctcc ccagcacttc ctacacagcc tgctgaagaa gcagcacgaa agagagaggt
     1321 tcgtctaatg aagaacaggg aagcagcaag agaatgtcgt agaaagaaga aagaatatgt
     1381 gaaatgttta gagaacagag tggcagtgct tgaaaaccaa aacaaaacat tgattgagga
     1441 gctaaaagca cttaaggacc tttactgcca caagtcagat taatttggga tccactagtt
     1501 ctagagggcc ctattctata gtgtcaccta aatgctagag gatctttgtg aaggaacctt
     1561 acttctgtgg tgtgacataa ttggacaaac tacctacaga gatttaaagc tctaaggtaa
     1621 atataaaatt tttaagtgta taatgtgtta aactactgat tctaattgtt tgtgtatttt
     1681 agattccaac ctatggaact gatgaatggg agcagtggtg gaatgccttt aatgaggaaa
     1741 acctgttttg ctcagaagaa atgccatcta gtgatgatga ggctactgct gactctcaac
     1801 attctactcc tccaaaaaag aagagaaagg tagaagaccc caaggacttt ccttcagaat
     1861 tgctaagttt tttgagtcat gctgtgttta gtaatagaac tcttgcttgc tttgctattt
     1921 acaccacaaa ggaaaaagct gcactgctat acaagaaaat tatggaaaaa tatttgatgt
     1981 atagtgcctt gactagagat cataatcagc cataccacat ttgtagaggt tttacttgct
     2041 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     2101 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2161 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2221 tcttatcatg tctggatcat cccgccatgg tatcaacgcc atatttctat ttacagtagg
     2281 gacctcttcg ttgtgtaggt accgctgtat tcctagggaa atagtagagg caccttgaac
     2341 tgtctgcatc agccatatag cccccgctgt tcgacttaca aacacaggca cagtactgac
     2401 aaacccatac acctcctctg aaatacccat agttgctagg gctgtctccg aactcattac
     2461 accctccaaa gtcagagctg taatttcgcc atcaagggca gcgagggctt ctccagataa
     2521 aatagcttct gccgagagtc ccgtaagggt agacacttca gctaatccct cgatgaggtc
     2581 tactagaata gtcagtgcgg ctcccatttt gaaaattcac ttacttgatc agcttcagaa
     2641 gatggcggag ggcctccaac acagtaattt tcctcccgac tcttaaaata gaaaatgtca
     2701 agtcagttaa gcaggaagtg gactaactga cgcagctggc cgtgcgacat cctcttttaa
     2761 ttagttgcta ggcaacgccc tccagagggc gtgtggtttt gcaagaggaa gcaaaagcct
     2821 ctccacccag gcctagaatg tttccaccca atcattacta tgacaacagc tgtttttttt
     2881 agtattaagc agaggccggg gacccctggg cccgcttact ctggagaaaa agaagagagg
     2941 cattgtagag gcttccagag gcaacttgtc aaaacaggac tgcttctatt tctgtcacac
     3001 tgtctggccc tgtcacaagg tccagcacct ccataccccc tttaataagc agtttgggaa
     3061 cgggtgcggg tcttactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3121 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     3181 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa
     3241 ttcggcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3301 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3361 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3421 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3481 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3541 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3601 tacctacagc gtgagcattg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3661 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3721 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3781 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcaag ctagcttcta
     3841 gctagaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3901 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagcc
     3961 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     4021 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggccgcccac tacgtgaacc
     4081 atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     4141 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     4201 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     4261 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg gttgctttga
     4321 cgagaccgta taacgtgctt tcctcgttgg aatcagagcg ggagctaaac aggaggccga
     4381 ttaaagggat tttagacagg aacggtacgc cagctggatt accaaagggc ctcgtgatac
     4441 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4501 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4561 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4621 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4681 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4741 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4801 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4861 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4921 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4981 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5041 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5101 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5161 ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5221 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5281 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5341 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5401 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5461 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5521 taaaacttca tttttaattt ctctagcgcg ttgacattga ttattgacta gttattaata
     5581 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     5641 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     5701 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggacta
     5761 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     5821 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     5881 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     5941 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     6001 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     6061 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     6121 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
     6181 taatacgact cactataggg agaccc
//
