LOCUS Exported 5604 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of mouse codon-optimized PhiC31 recombinase with bovine growth hormone polyadenylation (bpA) sequence.. ACCESSION . VERSION . KEYWORDS pPGKPhiC31obpA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5604) AUTHORS Raymond CS, Soriano P TITLE High-Efficiency FLP and PhiC31 Site-Specific Recombination in Mammalian Cells. JOURNAL PLoS ONE. 2007 . 2():e162. PUBMED 17225864 REFERENCE 2 (bases 1 to 5604) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5604 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 24..128 /gene="bla" /label=AmpR promoter CDS 129..989 /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(347..366) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1160..1748 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1649..1668 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1902..1919 /label=L4440 /note="L4440 vector, forward primer" protein_bind 2036..2057 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2072..2102 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2110..2126 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 2115..2137 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2134..2150 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2134..2150 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 2169..2189 /label=T3 /note="T3 promoter, forward primer" promoter 2171..2189 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind 2217..2233 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 2218..2234 /label=KS primer /note="common sequencing primer, one of multiple similar variants" promoter 2259..2758 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(2294..2317) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 2691..2710 /label=mPGK-F /note="Mouse PGK promoter, forward primer" regulatory 2768..2777 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2774..4588 /codon_start=1 /product="integrase from phage phi-C31" /label=phage phi-C31 integrase /translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI EKRASITWAKPPTDDDEDDAQDGTEDVAA" CDS 4589..4609 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind complement(4636..4653) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4642..4866 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(4960..4979) /label=T7 /note="T7 promoter, forward primer" promoter complement(4961..4979) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4989..5006) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4989..5005) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4998..5020) /label=M13/pUC Forward /note="In lacZ gene" rep_origin complement(5146..5601) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5283..5304) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5495..5514 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt 61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa 121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt 181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt 241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt 301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg 361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga 421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa 481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga 541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa 601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca 661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta 721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac 781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc 841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag 901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga 961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt 1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata 1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag 1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt 1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc 1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa 1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa 1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc 1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa 1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa 1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg 1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc 1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg 1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg 1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg 1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat 1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg 2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt 2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg 2161 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggtaccgggc cccccctcga 2221 ggtcgacggt atcgataagc ttgatatcga attctaccgg gtaggggagg cgcttttccc 2281 aaggcagtct ggagcatgcg ctttagcagc cccgctgggc acttggcgct acacaagtgg 2341 cctctggcct cgcacacatt ccacatccac cggtaggcgc caaccggctc cgttctttgg 2401 tggccccttc gcgccacctt ctactcctcc cctagtcagg aagttccccc ccgccccgca 2461 gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt ctcactagtc tcgtgcagat 2521 ggacagcacc gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc aatagcagct 2581 ttgctccttc gctttctggg ctcagaggct gggaaggggt gggtccgggg gcgggctcag 2641 gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac 2701 gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac 2761 catcgccgcc accatggata cctacgccgg agcctacgac agacagagcc gggagagaga 2821 gaacagcagc gccgccagcc ccgccaccca gagaagcgcc aacgaggata aggccgccga 2881 tctgcagaga gaggtggaga gggacggcgg cagattcaga tttgtgggcc acttcagcga 2941 ggcccctggc accagcgcct tcggcaccgc cgagagaccc gagttcgaga gaatcctgaa 3001 cgagtgtagg gccggcaggc tgaacatgat catcgtgtac gacgtgtccc ggttcagcag 3061 gctgaaggtg atggacgcca tccctatcgt gtccgagctg ctggccctgg gcgtgaccat 3121 cgtgtccacc caggaaggcg tctttagaca gggcaacgtg atggacctga tccacctgat 3181 catgaggctg gacgccagcc acaaggagag cagcctgaag agcgccaaga tcctggacac 3241 caagaacctg cagagggagc tgggcggcta tgtgggcggc aaggccccct acggcttcga 3301 gctggtgtcc gagaccaagg agatcacccg gaacggcagg atggtgaacg tggtgatcaa 3361 caagctggcc cacagcacca cccccctgac cggccccttc gagtttgagc ccgacgtgat 3421 caggtggtgg tggcgggaga tcaagaccca caagcacctg cctttcaagc ccggcagcca 3481 ggccgccatc caccccggca gcatcaccgg cctgtgtaag agaatggacg ccgacgccgt 3541 gcccaccaga ggcgagacca tcggcaagaa aaccgccagc agcgcctggg accccgccac 3601 cgtgatgaga atcctgaggg accctaggat cgccggcttc gccgccgagg tgatctacaa 3661 gaagaagccc gacggcaccc ccaccaccaa gatcgagggc tacagaatcc agagagaccc 3721 catcaccctg agacctgtgg agctggactg tggccctatc atcgagcctg ccgagtggta 3781 cgagctgcag gcctggctgg acggcagagg cagaggcaag ggcctgagca gaggccaggc 3841 catcctgagc gccatggaca agctgtactg tgagtgtggc gccgtgatga ccagcaagag 3901 aggcgaggag agcatcaagg acagctaccg gtgccggaga agaaaggtgg tggaccccag 3961 cgcccctggc cagcacgagg gcacctgtaa tgtgagcatg gccgccctgg acaagttcgt 4021 ggccgagcgg atcttcaaca agatccggca cgccgagggc gacgaggaga ccctggccct 4081 gctgtgggag gccgccagaa gattcggcaa gctgaccgag gcccccgaga agagcggcga 4141 gagggccaac ctggtggccg agagagccga cgccctgaac gccctggagg agctgtacga 4201 ggacagagcc gccggagcct atgacggccc tgtgggcagg aagcacttca gaaagcagca 4261 ggccgccctg accctgagac agcagggcgc cgaggaaaga ctggccgagc tggaggccgc 4321 cgaggcccct aagctgcccc tggatcagtg gttccccgag gatgccgacg ccgaccccac 4381 cggccccaag tcctggtggg gcagagccag cgtggacgac aagagggtgt tcgtgggcct 4441 gttcgtggat aagatcgtgg tgaccaagag caccaccggc aggggccagg gcacccccat 4501 cgagaagaga gccagcatca cctgggccaa gcctcccacc gacgacgacg aggatgacgc 4561 ccaggacggc accgaggacg tggccgcccc taagaaaaag cggaaagtgt gatagatcta 4621 gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct 4681 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg 4741 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc 4801 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct 4861 ctatggcttc tgaggcggaa agaaccagct ggggctcgag atccactagt tctagcctcg 4921 aggctagagc ggccgccacc gcggtggagc tccaattcgc cctatagtga gtcgtattac 4981 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa 5041 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc 5101 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggaaatt gtaagcgtta 5161 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg 5221 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg 5281 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa 5341 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg 5401 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt 5461 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg 5521 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta 5581 atgcgccgct acagggcgcg tcag //