LOCUS       sequence-197917-        6061 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cAMPr with a directly fused N-terminal 6x His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6061)
  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 6061)
  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-N-(His-6X)-cAMPr
FEATURES             Location/Qualifiers
     source          1..6061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..507
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(43..66)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     440..459
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      542..551
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(557..590)
                     /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(593..626)
                     /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     628..644
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     629..645
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             673..690
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     1953..1974
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2977..2993)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2977..2993)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    complement(3017..3041)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3067..3108
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3118..3135
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3162..3283
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3173..3192)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3227..3246
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3466..3921
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3553..3572)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3763..3784
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3988..4007)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4107..4129
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4167..4185)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4253..4357
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4358..4426,4427..5218)
                     /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(4576..4595)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5389..5977
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5878..5897
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc
       61 ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc
      121 ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc
      181 ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa
      241 gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta
      301 ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg
      361 ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga
      421 aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc
      481 tcctcttcct catctccggg cctttcgacc tgcatccatc tagatctcga aggatctgga
      541 ggccaccatg gtgtcgataa cttcgtatag catacattat acgaagttat cgataacttc
      601 gtataggata ctttatacga agttatctcg aggtcgacgg tatcgataag cttgatatcg
      661 aattccacca tgcatcatca ccaccaccat ggcaacgccg ccgccgccaa gaagggcagc
      721 gagcaggaga gcgtgaaaga gttcctagcc aaagccaagg aagatttcct gaaaaaatgg
      781 gagacccctt ctcagaatac agcccagttg gatcagtttg atagaatcaa gacccttggc
      841 accggctcct ttgggcgagt gatgctggtg aagcacaagg agagtgggaa ccactacgcc
      901 atgaagatct tagacaagca gaaggtggtg aagctaaagc agatcgagca cactctgaat
      961 gagaagcgca tcctgcaggc cgtcaacttc ccgttcctgg tcaaacttga attctccttc
     1021 aaggacaact caaacctgta catggtcatg gagtatgtag ctggtggcga gatgttctcc
     1081 cacctacggc ggattggaag gttcagcgag ccccatgccc gtttctacgc ggcgcagatc
     1141 gtcctgacct ttgagtatct gcactccctg gacctcatct accgggacct gaagcccgag
     1201 aatcttctca tcgaccagca gggctatatt caggtgacag acttcggttt tgccaagcgt
     1261 gtgaaaggcc gtacttggac cttgtgtggg acccctgagt acttggcccc cgagattatc
     1321 ctgagcaaag gctacaacaa ggctgtggac tggtgggctc tcggagtcct catctacgag
     1381 atggctgctg gttacccacc cttcttcgct gaccagccta tccagatcta tgagaaaatc
     1441 gtctctggga aggtgcggtt cccatcccac ttcagctctg acttgaagga cctgctgcgg
     1501 aaccttctgc aagtggatct caccaagcgc tttgggaacc tcaagaacgg ggtcaatgac
     1561 atcaagaacc acaagtggtt tgccacgact gactggattg ccatctatca gagaaaggtg
     1621 gaagctccct tcataccaaa gtttaaaggc cctggggaca cgagtaactt tgacgactat
     1681 gaggaggaag agatccgggt ctccatcaat gagaagtgtg gcaaggagtt tactgagttt
     1741 cccgggaacg tctatatcaa ggccgacgag cagaagaacg gcatcaaggc gtacttcaag
     1801 atccgccaca acatcgaggg cggcggcgtg cagctcgcct accactacca gcagaacacc
     1861 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcgt gcagtccatg
     1921 ctttcgaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1981 gccgggatca ctctcggcat ggacgagctg tacaagggcg gtaccggagg gagcgaatcc
     2041 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatccaggt cgagctggac
     2101 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gtgagggcga tgccacctac
     2161 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     2221 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     2281 cagcacgact tcttcaagtc cgccatgccc gaaggctaca tccaggagcg caccatcttc
     2341 ttcaagggcg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     2401 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     2461 aagctggagt acaacaccgc cccccccgca tgcaagggcc ggcggcgacg cggggccatc
     2521 agcgctgagg tctacaccga ggaggatgcc gcgtcctatg ttcggaaggt tataccgaaa
     2581 gattataaga caatggctgc tttagctaaa gccattgaaa agaacgtact gttttcacat
     2641 cttgatgata acgagagaag tgacattttt gacgccatgt tcccggtttc ctttattgct
     2701 ggagagactg ttattcagca gggtgacgaa ggggataact tctacgtgat tgaccaagga
     2761 gagatggacg tctatgtcaa caatgaatgg gcaaccagtg ttggggaagg agggagcttc
     2821 ggggagcttg ctctgattta cgggactcct cgagcggcca ctgtcaaggc caagacgaac
     2881 gtgaaactgt ggggcattga ccgggacagc tacagaagga tcctcatggg aagcacgctg
     2941 agaaagcgga agatgtaaaa ttcctgcagc ccgggggatc cactagttct agagcggccg
     3001 cagggtgggc gcgccgaccc agctttcttg tacaaagtgg ttgatctaga gggcccgcgg
     3061 ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg taccggtcat
     3121 catcaccatc accattgagc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     3181 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3241 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3301 tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct
     3361 ctacaaatgt ggtaaaatcg cgataaggat ccgggctggc gtaatagcga agaggcccgc
     3421 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg
     3481 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3541 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3601 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3661 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     3721 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3781 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     3841 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     3901 aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3961 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4021 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4081 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4141 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4201 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4261 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4321 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4381 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4441 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4501 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4561 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4621 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4681 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4741 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4801 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4861 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4921 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4981 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5041 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5101 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5161 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5221 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5281 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5341 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5401 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5461 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5521 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5581 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5641 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5701 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5761 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5821 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5881 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5941 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6001 acggttcctg gccttttgct ggccttttgc tcacatggct cgacagatct cgacctcgaa
     6061 a
//
