LOCUS       sequence-193967-        6042 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cAMPr with a point mutation Y205A which should render PKA-C 
            catalytically dead.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6042)
  AUTHORS   Hackley CR, Mazzoni EO, Blau J
  TITLE     cAMPr: A single-wavelength fluorescent sensor for cyclic AMP.
  JOURNAL   Sci Signal. 2018 Mar 6;11(520). pii: 11/520/eaah3738. doi: 
            10.1126/scisignal.aah3738.
  PUBMED    29511120
REFERENCE   2  (bases 1 to 6042)
  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: "Sci 
            Signal."; date: "2018-03-6"; volume: "11(520). pii: 11/520/eaah3738.
            doi"; pages: " 10.1126/scisignal.aah3738"
COMMENT     Sequence Label: p2lox-cAMPr Y205A
FEATURES             Location/Qualifiers
     source          1..6042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        19..518
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(54..77)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     451..470
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      553..562
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(568..601)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(604..637)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     639..655
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     640..656
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1942..1963
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2967..2983)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2967..2983)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    complement(3007..3031)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3057..3098
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3108..3125
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3152..3273
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3163..3182)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3217..3236
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3456..3911
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3543..3562)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3753..3774
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3980..3999)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4099..4121
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4159..4177)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4245..4349
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4350..4418,4419..5210)
                     /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     complement(4568..4587)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5381..5969
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5870..5889
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgacctcgaa attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg
       61 ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt
      121 ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt
      181 ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt
      241 gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat
      301 ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg
      361 ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg
      421 cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg
      481 ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcatccat ctagatctcg
      541 aaggatctgg aggccaccat ggtgtcgata acttcgtata gcatacatta tacgaagtta
      601 tcgataactt cgtataggat actttatacg aagttatctc gaggtcgacg gtatcgataa
      661 gcttgatatc gaattcatgg gcaacgccgc cgccgccaag aagggcagcg agcaggagag
      721 cgtgaaagag ttcctagcca aagccaagga agatttcctg aaaaaatggg agaccccttc
      781 tcagaataca gcccagttgg atcagtttga tagaatcaag acccttggca ccggctcctt
      841 tgggcgagtg atgctggtga agcacaagga gagtgggaac cactacgcca tgaagatctt
      901 agacaagcag aaggtggtga agctaaagca gatcgagcac actctgaatg agaagcgcat
      961 cctgcaggcc gtcaacttcc cgttcctggt caaacttgaa ttctccttca aggacaactc
     1021 aaacctgtac atggtcatgg agtatgtagc tggtggcgag atgttctccc acctacggcg
     1081 gattggaagg ttcagcgagc cccatgcccg tttctacgcg gcgcagatcg tcctgacctt
     1141 tgagtatctg cactccctgg acctcatcta ccgggacctg aagcccgaga atcttctcat
     1201 cgaccagcag ggctatattc aggtgacaga cttcggtttt gccaagcgtg tgaaaggccg
     1261 tacttggacc ttgtgtggga cccctgaggc cttggccccc gagattatcc tgagcaaagg
     1321 ctacaacaag gctgtggact ggtgggctct cggagtcctc atctacgaga tggctgctgg
     1381 ttacccaccc ttcttcgctg accagcctat ccagatctat gagaaaatcg tctctgggaa
     1441 ggtgcggttc ccatcccact tcagctctga cttgaaggac ctgctgcgga accttctgca
     1501 agtggatctc accaagcgct ttgggaacct caagaacggg gtcaatgaca tcaagaacca
     1561 caagtggttt gccacgactg actggattgc catctatcag agaaaggtgg aagctccctt
     1621 cataccaaag tttaaaggcc ctggggacac gagtaacttt gacgactatg aggaggaaga
     1681 gatccgggtc tccatcaatg agaagtgtgg caaggagttt actgagtttc ccgggaacgt
     1741 ctatatcaag gccgacgagc agaagaacgg catcaaggcg tacttcaaga tccgccacaa
     1801 catcgagggc ggcggcgtgc agctcgccta ccactaccag cagaacaccc ccatcggcga
     1861 cggccccgtg ctgctgcccg acaaccacta cctgagcgtg cagtccatgc tttcgaaaga
     1921 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1981 tctcggcatg gacgagctgt acaagggcgg taccggaggg agcgaatcca tggtgagcaa
     2041 gggcgaggag ctgttcaccg gggtggtgcc catccaggtc gagctggacg gcgacgtaaa
     2101 cggccacaag ttcagcgtgt ccggcgaggg tgagggcgat gccacctacg gcaagctgac
     2161 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     2221 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     2281 cttcaagtcc gccatgcccg aaggctacat ccaggagcgc accatcttct tcaagggcga
     2341 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     2401 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     2461 caacaccgcc ccccccgcat gcaagggccg gcggcgacgc ggggccatca gcgctgaggt
     2521 ctacaccgag gaggatgccg cgtcctatgt tcggaaggtt ataccgaaag attataagac
     2581 aatggctgct ttagctaaag ccattgaaaa gaacgtactg ttttcacatc ttgatgataa
     2641 cgagagaagt gacatttttg acgccatgtt cccggtttcc tttattgctg gagagactgt
     2701 tattcagcag ggtgacgaag gggataactt ctacgtgatt gaccaaggag agatggacgt
     2761 ctatgtcaac aatgaatggg caaccagtgt tggggaagga gggagcttcg gggagcttgc
     2821 tctgatttac gggactcctc gagcggccac tgtcaaggcc aagacgaacg tgaaactgtg
     2881 gggcattgac cgggacagct acagaaggat cctcatggga agcacgctga gaaagcggaa
     2941 gatgtaagaa ttcctgcagc ccgggggatc cactagttct agagcggccg cagggtgggc
     3001 gcgccgaccc agctttcttg tacaaagtgg ttgatctaga gggcccgcgg ttcgaaggta
     3061 agcctatccc taaccctctc ctcggtctcg attctacgcg taccggtcat catcaccatc
     3121 accattgagc ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     3181 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     3241 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     3301 tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     3361 ggtaaaatcg cgataaggat ccgggctggc gtaatagcga agaggcccgc accgatcgcc
     3421 cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag
     3481 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3541 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3601 tctaaatcgg gggctccctt tagggttccg atttagagct ttacggcacc tcgaccgcaa
     3661 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3721 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3781 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3841 ttggttaaaa aatgagctga tttaacaaat atttaacgcg aattttaaca aaatattaac
     3901 gtttacaatt tcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3961 gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     4021 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     4081 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     4141 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     4201 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4261 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4321 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4381 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4441 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4501 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4561 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4621 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4681 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4741 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4801 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4861 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     4921 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4981 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5041 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5101 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5161 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5221 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5281 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5341 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5401 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5461 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5521 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5581 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5641 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5701 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5761 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5821 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5881 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5941 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6001 tggccttttg ctggcctttt gctcacatgg ctcgacagat ct
//
