LOCUS       sequence-189838-        6043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses cAMPr in mammalian cells.
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
FEATURES             Location/Qualifiers
     source          1..6043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..526
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(62..85)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     459..478
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      561..570
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(576..609)
                     /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(612..645)
                     /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     647..663
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     648..664
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1954..1975
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2978..2994)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2978..2994)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    complement(3018..3042)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3068..3109
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3119..3136
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3163..3284
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3174..3193)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3228..3247
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3467..3922
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3554..3573)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3764..3785
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3989..4008)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4108..4130
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4168..4186)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4254..4358
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4359..4427,4428..5219)
                     /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(4577..4596)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5390..5978
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5879..5898
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acagatctcg acctcgaaat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg
       61 agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg
      121 cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc
      181 gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg
      241 caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc
      301 tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc
      361 tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag
      421 gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg
      481 cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct gcatccatct
      541 agatctcgaa ggatctggag gccaccatgg tgtcgataac ttcgtatagc atacattata
      601 cgaagttatc gataacttcg tataggatac tttatacgaa gttatctcga ggtcgacggt
      661 atcgataagc ttgatatcga attccaccat gggcaacgcc gccgccgcca agaagggcag
      721 cgagcaggag agcgtgaaag agttcctagc caaagccaag gaagatttcc tgaaaaaatg
      781 ggagacccct tctcagaata cagcccagtt ggatcagttt gatagaatca agacccttgg
      841 caccggctcc tttgggcgag tgatgctggt gaagcacaag gagagtggga accactacgc
      901 catgaagatc ttagacaagc agaaggtggt gaagctaaag cagatcgagc acactctgaa
      961 tgagaagcgc atcctgcagg ccgtcaactt cccgttcctg gtcaaacttg aattctcctt
     1021 caaggacaac tcaaacctgt acatggtcat ggagtatgta gctggtggcg agatgttctc
     1081 ccacctacgg cggattggaa ggttcagcga gccccatgcc cgtttctacg cggcgcagat
     1141 cgtcctgacc tttgagtatc tgcactccct ggacctcatc taccgggacc tgaagcccga
     1201 gaatcttctc atcgaccagc agggctatat tcaggtgaca gacttcggtt ttgccaagcg
     1261 tgtgaaaggc cgtacttgga ccttgtgtgg gacccctgag tacttggccc ccgagattat
     1321 cctgagcaaa ggctacaaca aggctgtgga ctggtgggct ctcggagtcc tcatctacga
     1381 gatggctgct ggttacccac ccttcttcgc tgaccagcct atccagatct atgagaaaat
     1441 cgtctctggg aaggtgcggt tcccatccca cttcagctct gacttgaagg acctgctgcg
     1501 gaaccttctg caagtggatc tcaccaagcg ctttgggaac ctcaagaacg gggtcaatga
     1561 catcaagaac cacaagtggt ttgccacgac tgactggatt gccatctatc agagaaaggt
     1621 ggaagctccc ttcataccaa agtttaaagg ccctggggac acgagtaact ttgacgacta
     1681 tgaggaggaa gagatccggg tctccatcaa tgagaagtgt ggcaaggagt ttactgagtt
     1741 tcccgggaac gtctatatca aggccgacga gcagaagaac ggcatcaagg cgtacttcaa
     1801 gatccgccac aacatcgagg gcggcggcgt gcagctcgcc taccactacc agcagaacac
     1861 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagcg tgcagtccat
     1921 gctttcgaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1981 cgccgggatc actctcggca tggacgagct gtacaagggc ggtaccggag ggagcgaatc
     2041 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatccagg tcgagctgga
     2101 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggtgagggcg atgccaccta
     2161 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     2221 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     2281 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac atccaggagc gcaccatctt
     2341 cttcaagggc gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2401 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2461 caagctggag tacaacaccg ccccccccgc atgcaagggc cggcggcgac gcggggccat
     2521 cagcgctgag gtctacaccg aggaggatgc cgcgtcctat gttcggaagg ttataccgaa
     2581 agattataag acaatggctg ctttagctaa agccattgaa aagaacgtac tgttttcaca
     2641 tcttgatgat aacgagagaa gtgacatttt tgacgccatg ttcccggttt cctttattgc
     2701 tggagagact gttattcagc agggtgacga aggggataac ttctacgtga ttgaccaagg
     2761 agagatggac gtctatgtca acaatgaatg ggcaaccagt gttggggaag gagggagctt
     2821 cggggagctt gctctgattt acgggactcc tcgagcggcc actgtcaagg ccaagacgaa
     2881 cgtgaaactg tggggcattg accgggacag ctacagaagg atcctcatgg gaagcacgct
     2941 gagaaagcgg aagatgtaaa attcctgcag cccgggggat ccactagttc tagagcggcc
     3001 gcagggtggg cgcgccgacc cagctttctt gtacaaagtg gttgatctag agggcccgcg
     3061 gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca
     3121 tcatcaccat caccattgag cggccgcttc gagcagacat gataagatac attgatgagt
     3181 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     3241 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     3301 ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc
     3361 tctacaaatg tggtaaaatc gcgataagga tccgggctgg cgtaatagcg aagaggcccg
     3421 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggacgc gccctgtagc
     3481 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3541 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3601 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3661 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3721 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3781 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3841 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     3901 aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3961 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4021 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4081 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4141 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     4201 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4261 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4321 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4381 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4441 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4501 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4561 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4621 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4681 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4741 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4801 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4861 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4921 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4981 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5041 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5101 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5161 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5221 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5281 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5341 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5401 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5461 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5521 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5581 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5641 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5701 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5761 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5821 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5881 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5941 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6001 tacggttcct ggccttttgc tggccttttg ctcacatggc tcg
//
