LOCUS Exported 6043 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses cAMPr in mammalian cells. ACCESSION . VERSION . KEYWORDS p2lox-cAMPr 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6043 /organism="synthetic DNA construct" /mol_type="other DNA" 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 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind 576..609 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 612..645 /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). 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 4359..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 //