LOCUS       sequence-228486-        6679 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 6679)
  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 6679)
  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 M1
FEATURES             Location/Qualifiers
     source          1..6679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             856..879
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(1998..2016)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1999..2016)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2036..2053)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2042..2266
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2312..2740
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2399..2418)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2609..2630
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2749..2769)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2754..3083
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2934..3069
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2996..3015
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3150..3944
                     /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(3204..3223)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3814..3833
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4118..4239
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4155..4174)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4209..4228
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4288..4304)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4288..4304)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4301..4323)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4312..4328
                     /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(4336..4342,4343..4360,4361..4366))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4381..4402
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4519..4536)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4690..5278)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4770..4789)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5449..6240,6241..6309))
                     /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     6072..6091
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6310..6414)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6489..6508)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctc
      661 cacaattcgg gcggcggctg ctgctgcctg tggcccgggc ggctgggaga agccgagtgt
      721 tggtgagtga cgcggcggag gtgtagtttg acgcggtgtg ttacgtgggg gagagaataa
      781 aactccagcg agatccgggc cgcgaacgaa agcagtgacg gaggagcttg taccaccggt
      841 gactagatga ccatggacta caaggacgac gatgacaagc tgcaggaatt cggatccgac
      901 tctggagcag acaaccagca gagtggagat gctgctgtaa cagaagctga aagtcaacaa
      961 atgacagttc aagcccagcc acagattgcc acattagccc aggtatccat gccagcagct
     1021 catgcgacgt catctgctcc cactgtaacc ttagtgcagc tgcccaatgg gcagacagtc
     1081 caggtccatg gggtcatcca ggcggcccag ccatcagtta ttcagtctcc acaagtccaa
     1141 acagttcagt cttcctgtaa ggacttaaaa agacttttct ccggaactca gatttcaact
     1201 attgcagaaa gtgaagattc acaggagtct gtggatagtg taactgattc ccaaaaacga
     1261 agggaaatcc tttcaaggag gcctgcctac aggaaaattt tgaatgactt atcttctgat
     1321 gcaccagggg tgccaaggat tgaagaagaa aaatcagaag aagagacttc agcccctgcc
     1381 atcaccactg taacagtgcc aaccccgatt taccaaacta gcagtgggca gtatattgcc
     1441 attacccagg gaggagcaat acagctggct aacaatggta ccgatggggt acagggcctg
     1501 cagacattaa ccatgaccaa tgcagctgcc actcagccgg gtactaccat tctacaatat
     1561 gcacagacca ctgatggaca gcagattcta gtgcccagca accaagttgt tgttcaagct
     1621 gcctctggtg atgtacaaac ataccagatt cgcacagcac ccactagcac cattgcccct
     1681 ggagttgtta tggcgtcctc cccagcactt cctacacagc ctgctgaaga agcagcacga
     1741 aagagagagg ttcgtctaat gaagaacagg gaagcagcaa gagaatgtcg tagaaagaag
     1801 aaagaatatg tgaaatgttt agagaacaga gtggcagtgc ttgaaaacca aaacaaaaca
     1861 ttgattgagg agctaaaagc acttaaggac ctttactgcc acaagtcaga ttaatttggg
     1921 atttaaattt tcacctattg tggtggaaaa tggactggat tgggggtacc gtcgagcatg
     1981 catctagagg gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg
     2041 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2101 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2161 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2221 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     2281 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     2341 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     2401 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2461 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2521 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     2581 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     2641 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     2701 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     2761 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2821 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2881 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     2941 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3001 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     3061 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     3121 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3181 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3241 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3301 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     3361 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3421 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3481 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3541 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     3601 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     3661 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     3721 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     3781 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     3841 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     3901 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     3961 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4021 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4081 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4141 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4201 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     4261 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4321 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4381 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4441 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4501 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4561 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4621 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4681 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4741 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4801 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4861 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4921 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4981 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5041 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5101 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5161 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5221 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5281 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5341 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5401 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5461 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5521 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5581 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5641 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5701 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5761 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5821 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5881 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5941 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6001 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6061 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6121 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6181 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6241 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6301 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6361 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6421 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     6481 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     6541 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     6601 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     6661 cgggccagat atacgcgtt
//
