LOCUS       sequence-197911-        6043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cAMPr with a point mutation R626A which theoretically decreases its 
            affinity for cAMP.
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 R626A
FEATURES             Location/Qualifiers
     source          1..6043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..502
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(38..61)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     435..454
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      537..546
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(552..585)
                     /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(588..621)
                     /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     623..639
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     624..640
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1930..1951
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2954..2970)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2954..2970)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    complement(2994..3018)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3044..3085
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3095..3112
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3139..3260
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3150..3169)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3204..3223
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3443..3898
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3530..3549)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3740..3761
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3965..3984)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4084..4106
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4144..4162)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4230..4334
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4335..4403,4404..5195)
                     /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(4553..4572)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5366..5954
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5855..5874
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct
       61 gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag
      121 gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt
      181 caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc
      241 acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct
      301 ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag
      361 gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc
      421 ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc
      481 ttcctcatct ccgggccttt cgacctgcat ccatctagat ctcgaaggat ctggaggcca
      541 ccatggtgtc gataacttcg tatagcatac attatacgaa gttatcgata acttcgtata
      601 ggatacttta tacgaagtta tctcgaggtc gacggtatcg ataagcttga tatcgaattc
      661 caccatgggc aacgccgccg ccgccaagaa gggcagcgag caggagagcg tgaaagagtt
      721 cctagccaaa gccaaggaag atttcctgaa aaaatgggag accccttctc agaatacagc
      781 ccagttggat cagtttgata gaatcaagac ccttggcacc ggctcctttg ggcgagtgat
      841 gctggtgaag cacaaggaga gtgggaacca ctacgccatg aagatcttag acaagcagaa
      901 ggtggtgaag ctaaagcaga tcgagcacac tctgaatgag aagcgcatcc tgcaggccgt
      961 caacttcccg ttcctggtca aacttgaatt ctccttcaag gacaactcaa acctgtacat
     1021 ggtcatggag tatgtagctg gtggcgagat gttctcccac ctacggcgga ttggaaggtt
     1081 cagcgagccc catgcccgtt tctacgcggc gcagatcgtc ctgacctttg agtatctgca
     1141 ctccctggac ctcatctacc gggacctgaa gcccgagaat cttctcatcg accagcaggg
     1201 ctatattcag gtgacagact tcggttttgc caagcgtgtg aaaggccgta cttggacctt
     1261 gtgtgggacc cctgagtact tggcccccga gattatcctg agcaaaggct acaacaaggc
     1321 tgtggactgg tgggctctcg gagtcctcat ctacgagatg gctgctggtt acccaccctt
     1381 cttcgctgac cagcctatcc agatctatga gaaaatcgtc tctgggaagg tgcggttccc
     1441 atcccacttc agctctgact tgaaggacct gctgcggaac cttctgcaag tggatctcac
     1501 caagcgcttt gggaacctca agaacggggt caatgacatc aagaaccaca agtggtttgc
     1561 cacgactgac tggattgcca tctatcagag aaaggtggaa gctcccttca taccaaagtt
     1621 taaaggccct ggggacacga gtaactttga cgactatgag gaggaagaga tccgggtctc
     1681 catcaatgag aagtgtggca aggagtttac tgagtttccc gggaacgtct atatcaaggc
     1741 cgacgagcag aagaacggca tcaaggcgta cttcaagatc cgccacaaca tcgagggcgg
     1801 cggcgtgcag ctcgcctacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1861 gctgcccgac aaccactacc tgagcgtgca gtccatgctt tcgaaagacc ccaacgagaa
     1921 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     1981 cgagctgtac aagggcggta ccggagggag cgaatccatg gtgagcaagg gcgaggagct
     2041 gttcaccggg gtggtgccca tccaggtcga gctggacggc gacgtaaacg gccacaagtt
     2101 cagcgtgtcc ggcgagggtg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     2161 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     2221 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     2281 catgcccgaa ggctacatcc aggagcgcac catcttcttc aagggcgacg gcaactacaa
     2341 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2401 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca acaccgcccc
     2461 ccccgcatgc aagggccggc ggcgacgcgg ggccatcagc gctgaggtct acaccgagga
     2521 ggatgccgcg tcctatgttg ctaaggttat accgaaagat tataagacaa tggctgcttt
     2581 agctaaagcc attgaaaaga acgtactgtt ttcacatctt gatgataacg agagaagtga
     2641 catttttgac gccatgttcc cggtttcctt tattgctgga gagactgtta ttcagcaggg
     2701 tgacgaaggg gataacttct acgtgattga ccaaggagag atggacgtct atgtcaacaa
     2761 tgaatgggca accagtgttg gggaaggagg gagcttcggg gagcttgctc tgatttacgg
     2821 gactcctcga gcggccactg tcaaggccaa gacgaacgtg aaactgtggg gcattgaccg
     2881 ggacagctac agaaggatcc tcatgggaag cacgctgaga aagcggaaga tgtaaaattc
     2941 ctgcagcccg ggggatccac tagttctaga gcggccgcag ggtgggcgcg ccgacccagc
     3001 tttcttgtac aaagtggttg atctagaggg cccgcggttc gaaggtaagc ctatccctaa
     3061 ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat caccatcacc attgagcggc
     3121 cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     3181 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     3241 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     3301 gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgcga
     3361 taaggatccg ggctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     3421 gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3481 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3541 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3601 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3661 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3721 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3781 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3841 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc
     3901 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     3961 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     4021 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     4081 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     4141 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     4201 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4261 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4321 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4381 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4441 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4501 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4561 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4621 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4681 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4741 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4801 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4861 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4921 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4981 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5041 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5101 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5161 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5221 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5281 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5341 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5401 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5461 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5521 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5581 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5641 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5701 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5761 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5821 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5881 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5941 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6001 cttttgctca catggctcga cagatctcga cctcgaaatt cta
//
