LOCUS       sequence-193966-        6060 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 6060)
  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 6060)
  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..6060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        19..518
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(54..77)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     451..470
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      553..562
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(568..601)
                     /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(604..637)
                     /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     639..655
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     640..656
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             680..697
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     1960..1981
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2985..3001)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2985..3001)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    complement(3025..3049)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3075..3116
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3126..3143
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3170..3291
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3181..3200)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3235..3254
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3474..3929
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3561..3580)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3771..3792
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3998..4017)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4117..4139
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4177..4195)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4263..4367
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4368..4436,4437..5228)
                     /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(4586..4605)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5399..5987
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5888..5907
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgacctcgaa attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg
       61 ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt
      121 ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt
      181 ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt
      241 gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat
      301 ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg
      361 ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg
      421 cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg
      481 ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcatccat ctagatctcg
      541 aaggatctgg aggccaccat ggtgtcgata acttcgtata gcatacatta tacgaagtta
      601 tcgataactt cgtataggat actttatacg aagttatctc gaggtcgacg gtatcgataa
      661 gcttgatatc gaattcatgc atcatcacca ccaccatggc aacgccgccg ccgccaagaa
      721 gggcagcgag caggagagcg tgaaagagtt cctagccaaa gccaaggaag atttcctgaa
      781 aaaatgggag accccttctc agaatacagc ccagttggat cagtttgata gaatcaagac
      841 ccttggcacc ggctcctttg ggcgagtgat gctggtgaag cacaaggaga gtgggaacca
      901 ctacgccatg aagatcttag acaagcagaa ggtggtgaag ctaaagcaga tcgagcacac
      961 tctgaatgag aagcgcatcc tgcaggccgt caacttcccg ttcctggtca aacttgaatt
     1021 ctccttcaag gacaactcaa acctgtacat ggtcatggag tatgtagctg gtggcgagat
     1081 gttctcccac ctacggcgga ttggaaggtt cagcgagccc catgcccgtt tctacgcggc
     1141 gcagatcgtc ctgacctttg agtatctgca ctccctggac ctcatctacc gggacctgaa
     1201 gcccgagaat cttctcatcg accagcaggg ctatattcag gtgacagact tcggttttgc
     1261 caagcgtgtg aaaggccgta cttggacctt gtgtgggacc cctgagtact tggcccccga
     1321 gattatcctg agcaaaggct acaacaaggc tgtggactgg tgggctctcg gagtcctcat
     1381 ctacgagatg gctgctggtt acccaccctt cttcgctgac cagcctatcc agatctatga
     1441 gaaaatcgtc tctgggaagg tgcggttccc atcccacttc agctctgact tgaaggacct
     1501 gctgcggaac cttctgcaag tggatctcac caagcgcttt gggaacctca agaacggggt
     1561 caatgacatc aagaaccaca agtggtttgc cacgactgac tggattgcca tctatcagag
     1621 aaaggtggaa gctcccttca taccaaagtt taaaggccct ggggacacga gtaactttga
     1681 cgactatgag gaggaagaga tccgggtctc catcaatgag aagtgtggca aggagtttac
     1741 tgagtttccc gggaacgtct atatcaaggc cgacgagcag aagaacggca tcaaggcgta
     1801 cttcaagatc cgccacaaca tcgagggcgg cggcgtgcag ctcgcctacc actaccagca
     1861 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcgtgca
     1921 gtccatgctt tcgaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1981 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggcggta ccggagggag
     2041 cgaatccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tccaggtcga
     2101 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggtg agggcgatgc
     2161 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     2221 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     2281 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacatcc aggagcgcac
     2341 catcttcttc aagggcgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     2401 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     2461 ggggcacaag ctggagtaca acaccgcccc ccccgcatgc aagggccggc ggcgacgcgg
     2521 ggccatcagc gctgaggtct acaccgagga ggatgccgcg tcctatgttc ggaaggttat
     2581 accgaaagat tataagacaa tggctgcttt agctaaagcc attgaaaaga acgtactgtt
     2641 ttcacatctt gatgataacg agagaagtga catttttgac gccatgttcc cggtttcctt
     2701 tattgctgga gagactgtta ttcagcaggg tgacgaaggg gataacttct acgtgattga
     2761 ccaaggagag atggacgtct atgtcaacaa tgaatgggca accagtgttg gggaaggagg
     2821 gagcttcggg gagcttgctc tgatttacgg gactcctcga gcggccactg tcaaggccaa
     2881 gacgaacgtg aaactgtggg gcattgaccg ggacagctac agaaggatcc tcatgggaag
     2941 cacgctgaga aagcggaaga tgtaagaatt cctgcagccc gggggatcca ctagttctag
     3001 agcggccgca gggtgggcgc gccgacccag ctttcttgta caaagtggtt gatctagagg
     3061 gcccgcggtt cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta
     3121 ccggtcatca tcaccatcac cattgagcgg ccgcttcgag cagacatgat aagatacatt
     3181 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     3241 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     3301 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag
     3361 taaaacctct acaaatgtgg taaaatcgcg ataaggatcc gggctggcgt aatagcgaag
     3421 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc
     3481 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3541 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3601 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagagcttt
     3661 acggcacctc gaccgcaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     3721 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3781 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3841 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaatat ttaacgcgaa
     3901 ttttaacaaa atattaacgt ttacaatttc gcctgatgcg gtattttctc cttacgcatc
     3961 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     4021 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     4081 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     4141 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     4201 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     4261 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4321 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4381 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4441 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4501 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4561 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4621 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4681 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4741 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4801 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4861 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4921 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4981 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5041 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5101 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5161 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5221 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5281 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5341 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5401 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5461 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5521 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     5581 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5641 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5701 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5761 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5821 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5881 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5941 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6001 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatggct cgacagatct
//
