LOCUS       sequence-197915-        6043 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 6043)
  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 6043)
  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..6043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        14..513
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(49..72)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     446..465
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      548..557
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(563..596)
                     /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(599..632)
                     /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     634..650
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     635..651
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1941..1962
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2965..2981)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2965..2981)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    complement(3005..3029)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3055..3096
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3106..3123
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3150..3271
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3161..3180)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3215..3234
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3454..3909
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3541..3560)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3751..3772
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3976..3995)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4095..4117
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4155..4173)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4241..4345
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4346..4414,4415..5206)
                     /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(4564..4583)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5377..5965
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5866..5885
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcgaaattct accgggtagg ggaggcgctt ttcccaaggc agtctggagc atgcgcttta
       61 gcagccccgc tgggcacttg gcgctacaca agtggcctct ggcctcgcac acattccaca
      121 tccaccggta ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact
      181 cctcccctag tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa
      241 atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag
      301 cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag
      361 aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc
      421 gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg
      481 ctgttctcct cttcctcatc tccgggcctt tcgacctgca tccatctaga tctcgaagga
      541 tctggaggcc accatggtgt cgataacttc gtatagcata cattatacga agttatcgat
      601 aacttcgtat aggatacttt atacgaagtt atctcgaggt cgacggtatc gataagcttg
      661 atatcgaatt ccaccatggg caacgccgcc gccgccaaga agggcagcga gcaggagagc
      721 gtgaaagagt tcctagccaa agccaaggaa gatttcctga aaaaatggga gaccccttct
      781 cagaatacag cccagttgga tcagtttgat agaatcaaga cccttggcac cggctccttt
      841 gggcgagtga tgctggtgaa gcacaaggag agtgggaacc actacgccat gaagatctta
      901 gacaagcaga aggtggtgaa gctaaagcag atcgagcaca ctctgaatga gaagcgcatc
      961 ctgcaggccg tcaacttccc gttcctggtc aaacttgaat tctccttcaa ggacaactca
     1021 aacctgtaca tggtcatgga gtatgtagct ggtggcgaga tgttctccca cctacggcgg
     1081 attggaaggt tcagcgagcc ccatgcccgt ttctacgcgg cgcagatcgt cctgaccttt
     1141 gagtatctgc actccctgga cctcatctac cgggacctga agcccgagaa tcttctcatc
     1201 gaccagcagg gctatattca ggtgacagac ttcggttttg ccaagcgtgt gaaaggccgt
     1261 acttggacct tgtgtgggac ccctgaggcc ttggcccccg agattatcct gagcaaaggc
     1321 tacaacaagg ctgtggactg gtgggctctc ggagtcctca tctacgagat ggctgctggt
     1381 tacccaccct tcttcgctga ccagcctatc cagatctatg agaaaatcgt ctctgggaag
     1441 gtgcggttcc catcccactt cagctctgac ttgaaggacc tgctgcggaa ccttctgcaa
     1501 gtggatctca ccaagcgctt tgggaacctc aagaacgggg tcaatgacat caagaaccac
     1561 aagtggtttg ccacgactga ctggattgcc atctatcaga gaaaggtgga agctcccttc
     1621 ataccaaagt ttaaaggccc tggggacacg agtaactttg acgactatga ggaggaagag
     1681 atccgggtct ccatcaatga gaagtgtggc aaggagttta ctgagtttcc cgggaacgtc
     1741 tatatcaagg ccgacgagca gaagaacggc atcaaggcgt acttcaagat ccgccacaac
     1801 atcgagggcg gcggcgtgca gctcgcctac cactaccagc agaacacccc catcggcgac
     1861 ggccccgtgc tgctgcccga caaccactac ctgagcgtgc agtccatgct ttcgaaagac
     1921 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     1981 ctcggcatgg acgagctgta caagggcggt accggaggga gcgaatccat ggtgagcaag
     2041 ggcgaggagc tgttcaccgg ggtggtgccc atccaggtcg agctggacgg cgacgtaaac
     2101 ggccacaagt tcagcgtgtc cggcgagggt gagggcgatg ccacctacgg caagctgacc
     2161 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2221 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     2281 ttcaagtccg ccatgcccga aggctacatc caggagcgca ccatcttctt caagggcgac
     2341 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2401 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2461 aacaccgccc cccccgcatg caagggccgg cggcgacgcg gggccatcag cgctgaggtc
     2521 tacaccgagg aggatgccgc gtcctatgtt cggaaggtta taccgaaaga ttataagaca
     2581 atggctgctt tagctaaagc cattgaaaag aacgtactgt tttcacatct tgatgataac
     2641 gagagaagtg acatttttga cgccatgttc ccggtttcct ttattgctgg agagactgtt
     2701 attcagcagg gtgacgaagg ggataacttc tacgtgattg accaaggaga gatggacgtc
     2761 tatgtcaaca atgaatgggc aaccagtgtt ggggaaggag ggagcttcgg ggagcttgct
     2821 ctgatttacg ggactcctcg agcggccact gtcaaggcca agacgaacgt gaaactgtgg
     2881 ggcattgacc gggacagcta cagaaggatc ctcatgggaa gcacgctgag aaagcggaag
     2941 atgtaaaatt cctgcagccc gggggatcca ctagttctag agcggccgca gggtgggcgc
     3001 gccgacccag ctttcttgta caaagtggtt gatctagagg gcccgcggtt cgaaggtaag
     3061 cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca tcaccatcac
     3121 cattgagcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     3181 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     3241 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     3301 tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     3361 taaaatcgcg ataaggatcc gggctggcgt aatagcgaag aggcccgcac cgatcgccct
     3421 tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg
     3481 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3541 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3601 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3661 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3721 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3781 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     3841 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     3901 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3961 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4021 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4081 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4141 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4201 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4261 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4321 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4381 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4441 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4501 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4561 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4621 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4681 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4741 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4801 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4861 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4921 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4981 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5041 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5101 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5161 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5221 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5281 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5341 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5401 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5461 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5521 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5581 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5641 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5701 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5761 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5821 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5881 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5941 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6001 cttttgctgg ccttttgctc acatggctcg acagatctcg acc
//
