LOCUS       sequence-154439-        6682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6682)
  AUTHORS   Du K, Asahara H, Jhala US, Wagner BL, Montminy M
  TITLE     Characterization of a CREB gain-of-function mutant with constitutive
            transcriptional activity in vivo.
  JOURNAL   Mol Cell Biol. 2000 Jun . 20(12):4320-7.
  PUBMED    10825195
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: "Mol Cell 
            Biol. 2000 Jun . 20(12):4320-7."
COMMENT     Sequence Label: pCF CREB
FEATURES             Location/Qualifiers
     source          1..6682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        156..535
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        536..739
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     690..710
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     784..803
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        784..802
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1011..1034
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2156..2174)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2157..2174)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2194..2211)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2200..2424
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2470..2898
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2557..2576)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2767..2788
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2907..2927)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2912..3241
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3092..3227
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3154..3173
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3308..4102
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3362..3381)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3972..3991
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4276..4397
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4313..4332)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4367..4386
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4446..4462)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4446..4462)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4459..4481)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4470..4486
                     /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)."
     promoter        complement(join(4494..4500,4501..4518,4519..4524))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4539..4560
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4677..4694)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4848..5436)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4928..4947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5607..6398,6399..6467))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6230..6249
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6468..6572)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6647..6666)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
       61 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
      121 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
      181 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      241 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      301 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      361 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      421 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      481 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      541 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      601 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      661 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      721 ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga
      781 aattaatacg actcactata gggagaccca agctccacaa ttcgggcggc ggctgctgct
      841 gcctgtggcc cgggcggctg ggagaagccg agtgttggtg agtgacgcgg cggaggtgta
      901 gtttgacgcg gtgtgttacg tgggggagag aataaaactc cagcgagatc cgggccgcga
      961 acgaaagcag tgacggagga gcttgtacca ccggtgacta gatgaccatg gactacaagg
     1021 acgacgatga caagctgcag gaattcatga ccatggactc tggagcagac aaccagcaga
     1081 gtggagatgc tgctgtaaca gaagctgaaa gtcaacaaat gacagttcaa gcccagccac
     1141 agattgccac attagcccag gtatccatgc cagcagctca tgcgacgtca tctgctccca
     1201 ctgtaacctt agtgcagctg cccaatgggc agacagtcca ggtccatggg gtcatccagg
     1261 cggcccagcc atcagttatt cagtctccac aagtccaaac agttcagtct tcctgtaagg
     1321 acttaaaaag acttttctcc ggaactcaga tttcaactat tgcagaaagt gaagattcac
     1381 aggagtctgt ggatagtgta actgattccc aaaaacgaag ggaaatcctt tcaaggaggc
     1441 cttcctacag gaaaattttg aatgacttat cttctgatgc accaggggtg ccaaggattg
     1501 aagaagaaaa atcagaagaa gagacttcag cccctgccat caccactgta acagtgccaa
     1561 ccccgattta ccaaactagc agtgggcagt atattgccat tacccaggga ggagcaatac
     1621 agctggctaa caatggtacc gatggggtac agggcctgca gacattaacc atgaccaatg
     1681 cagctgccac tcagccgggt actaccattc tacaatatgc acagaccact gatggacagc
     1741 agattctagt gcccagcaac caagttgttg ttcaagctgc ctctggtgat gtacaaacat
     1801 accagattcg cacagcaccc actagcacca ttgcccctgg agttgttatg gcgtcctccc
     1861 cagcacttcc tacacagcct gctgaagaag cagcacgaaa gagagaggtt cgtctaatga
     1921 agaacaggga agcagcaaga gaatgtcgta gaaagaagaa agaatatgtg aaatgtttag
     1981 agaacagagt ggcagtgctt gaaaaccaaa acaaaacatt gattgaggag ctaaaagcac
     2041 ttaaggacct ttactgccac aagtcagatt aatttgggat ttaaattttc acctattgtg
     2101 gtggaaaatg gactggattg ggggtaccgt cgagcatgca tctagagggc cctattctat
     2161 agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
     2221 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     2281 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     2341 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     2401 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     2461 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2521 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     2581 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     2641 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     2701 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     2761 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     2821 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     2881 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2941 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3001 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3061 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3121 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3181 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3241 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     3301 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3361 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3421 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3481 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3541 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3601 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3661 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3721 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     3781 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     3841 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     3901 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     3961 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4021 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4081 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4141 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4201 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4261 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4321 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4381 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     4441 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4501 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4561 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4621 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4681 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4741 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4801 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4861 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4921 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4981 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5041 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5101 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5161 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5221 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5281 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5341 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5401 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5461 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5521 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5581 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5641 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5701 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5761 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5821 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5881 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5941 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6001 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6061 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6121 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6181 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6241 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6301 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6361 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6421 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6481 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6541 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6601 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     6661 atgccgcata gttaagccag ta
//
