LOCUS       sequence-197918-        6043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cAMPr with a point mutation R739A which theoretically increases 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 R739A
FEATURES             Location/Qualifiers
     source          1..6043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..509
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(45..68)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     442..461
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      544..553
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(559..592)
                     /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(595..628)
                     /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     630..646
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     631..647
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1937..1958
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2961..2977)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2961..2977)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    complement(3001..3025)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3051..3092
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3102..3119
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3146..3267
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3157..3176)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3211..3230
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3450..3905
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3537..3556)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3747..3768
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3972..3991)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4091..4113
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4151..4169)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4237..4341
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4342..4410,4411..5202)
                     /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(4560..4579)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5373..5961
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5862..5881
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aattctaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag
       61 ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca
      121 ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc
      181 ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg
      241 aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg
      301 taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc
      361 tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc
      421 gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt
      481 tctcctcttc ctcatctccg ggcctttcga cctgcatcca tctagatctc gaaggatctg
      541 gaggccacca tggtgtcgat aacttcgtat agcatacatt atacgaagtt atcgataact
      601 tcgtatagga tactttatac gaagttatct cgaggtcgac ggtatcgata agcttgatat
      661 cgaattccac catgggcaac gccgccgccg ccaagaaggg cagcgagcag gagagcgtga
      721 aagagttcct agccaaagcc aaggaagatt tcctgaaaaa atgggagacc ccttctcaga
      781 atacagccca gttggatcag tttgatagaa tcaagaccct tggcaccggc tcctttgggc
      841 gagtgatgct ggtgaagcac aaggagagtg ggaaccacta cgccatgaag atcttagaca
      901 agcagaaggt ggtgaagcta aagcagatcg agcacactct gaatgagaag cgcatcctgc
      961 aggccgtcaa cttcccgttc ctggtcaaac ttgaattctc cttcaaggac aactcaaacc
     1021 tgtacatggt catggagtat gtagctggtg gcgagatgtt ctcccaccta cggcggattg
     1081 gaaggttcag cgagccccat gcccgtttct acgcggcgca gatcgtcctg acctttgagt
     1141 atctgcactc cctggacctc atctaccggg acctgaagcc cgagaatctt ctcatcgacc
     1201 agcagggcta tattcaggtg acagacttcg gttttgccaa gcgtgtgaaa ggccgtactt
     1261 ggaccttgtg tgggacccct gagtacttgg cccccgagat tatcctgagc aaaggctaca
     1321 acaaggctgt ggactggtgg gctctcggag tcctcatcta cgagatggct gctggttacc
     1381 cacccttctt cgctgaccag cctatccaga tctatgagaa aatcgtctct gggaaggtgc
     1441 ggttcccatc ccacttcagc tctgacttga aggacctgct gcggaacctt ctgcaagtgg
     1501 atctcaccaa gcgctttggg aacctcaaga acggggtcaa tgacatcaag aaccacaagt
     1561 ggtttgccac gactgactgg attgccatct atcagagaaa ggtggaagct cccttcatac
     1621 caaagtttaa aggccctggg gacacgagta actttgacga ctatgaggag gaagagatcc
     1681 gggtctccat caatgagaag tgtggcaagg agtttactga gtttcccggg aacgtctata
     1741 tcaaggccga cgagcagaag aacggcatca aggcgtactt caagatccgc cacaacatcg
     1801 agggcggcgg cgtgcagctc gcctaccact accagcagaa cacccccatc ggcgacggcc
     1861 ccgtgctgct gcccgacaac cactacctga gcgtgcagtc catgctttcg aaagacccca
     1921 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1981 gcatggacga gctgtacaag ggcggtaccg gagggagcga atccatggtg agcaagggcg
     2041 aggagctgtt caccggggtg gtgcccatcc aggtcgagct ggacggcgac gtaaacggcc
     2101 acaagttcag cgtgtccggc gagggtgagg gcgatgccac ctacggcaag ctgaccctga
     2161 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2221 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     2281 agtccgccat gcccgaaggc tacatccagg agcgcaccat cttcttcaag ggcgacggca
     2341 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2401 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaaca
     2461 ccgccccccc cgcatgcaag ggccggcggc gacgcggggc catcagcgct gaggtctaca
     2521 ccgaggagga tgccgcgtcc tatgttcgga aggttatacc gaaagattat aagacaatgg
     2581 ctgctttagc taaagccatt gaaaagaacg tactgttttc acatcttgat gataacgaga
     2641 gaagtgacat ttttgacgcc atgttcccgg tttcctttat tgctggagag actgttattc
     2701 agcagggtga cgaaggggat aacttctacg tgattgacca aggagagatg gacgtctatg
     2761 tcaacaatga atgggcaacc agtgttgggg aaggagggag cttcggggag cttgctctga
     2821 tttacgggac tcctcgagcg gccactgtca aggccaagac gaacgtgaaa ctgtggggca
     2881 ttgacgctga cagctacaga aggatcctca tgggaagcac gctgagaaag cggaagatgt
     2941 aaaattcctg cagcccgggg gatccactag ttctagagcg gccgcagggt gggcgcgccg
     3001 acccagcttt cttgtacaaa gtggttgatc tagagggccc gcggttcgaa ggtaagccta
     3061 tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg tcatcatcac catcaccatt
     3121 gagcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     3181 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     3241 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     3301 gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa
     3361 atcgcgataa ggatccgggc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     3421 aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc
     3481 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3541 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3601 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3661 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3721 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3781 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3841 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     3901 aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     3961 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4021 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4081 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4141 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4201 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4261 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4321 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4381 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4441 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4501 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4561 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4621 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4681 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4741 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4801 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4861 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     4921 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4981 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5041 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5101 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5161 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5221 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5281 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5341 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5401 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5461 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5521 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5581 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5641 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5701 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5761 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5821 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5881 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5941 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6001 tgctggcctt ttgctcacat ggctcgacag atctcgacct cga
//
