LOCUS Exported 5348 bp ds-DNA linear SYN 28-MAY-2021 DEFINITION Expresses FlpO under pCAG. ACCESSION . VERSION . KEYWORDS pCAG-FlpO SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5348) AUTHORS Weinberg BH, Pham NT, Caraballo LD, Lozanoski T, Engel A, Bhatia S, Wong WW TITLE Large-scale design of robust genetic circuits with multiple inputs and outputs for mammalian cells. JOURNAL Nat Biotechnol. 2017 Mar 27. doi: 10.1038/nbt.3805. PUBMED 28346402 REFERENCE 2 (bases 1 to 5348) AUTHORS . TITLE Direct Submission JOURNAL Exported May 28, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5348 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 346..368 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 430..449 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" regulatory 500..509 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 506..1804 /codon_start=1 /product="nuclear-targeted site-specific recombinase" /label=FLPo /note="FLPo is a mouse codon-optimized fusion between the SV40 nuclear localization signal and a thermostable version of the Saccharomyces cerevisiae site-specific recombinase FLP. Recombination occurs at FRT sequences." /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI" primer_bind complement(1890..1909) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 1955..2010 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(2009..2028) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(2371..2387) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2371..2387) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2384..2406) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2395..2411 /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)." promoter complement(2419..2449) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2464..2485 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin 2570..2704 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2631..2650 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 2724..2858 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2761..2780) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2815..2834 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(2926..2943) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3097..3685) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3177..3196) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3856..4716) /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 4479..4498 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4717..4821) /gene="bla" /label=AmpR promoter enhancer 4852..5231 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" ORIGIN 1 gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg 61 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga 121 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc 181 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg 241 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct 301 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc 361 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac 421 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattgatta 481 attcgagcga acgcgtgccg ccaccatggc tcctaagaag aagaggaagg tgatgagcca 541 gttcgacatc ctgtgcaaga ccccccccaa ggtgctggtg cggcagttcg tggagagatt 601 cgagaggccc agcggcgaga agatcgccag ctgtgccgcc gagctgacct acctgtgctg 661 gatgatcacc cacaacggca ccgccatcaa gagggccacc ttcatgagct acaacaccat 721 catcagcaac agcctgagct tcgacatcgt gaacaagagc ctgcagttca agtacaagac 781 ccagaaggcc accatcctgg aggccagcct gaagaagctg atccccgcct gggagttcac 841 catcatccct tacaacggcc agaagcacca gagcgacatc accgacatcg tgtccagcct 901 gcagctgcag ttcgagagca gcgaggaggc cgacaagggc aacagccaca gcaagaagat 961 gctgaaggcc ctgctgtccg agggcgagag catctgggag atcaccgaga agatcctgaa 1021 cagcttcgag tacaccagca ggttcaccaa gaccaagacc ctgtaccagt tcctgttcct 1081 ggccacattc atcaactgcg gcaggttcag cgacatcaag aacgtggacc ccaagagctt 1141 caagctggtg cagaacaagt acctgggcgt gatcattcag tgcctggtga ccgagaccaa 1201 gacaagcgtg tccaggcaca tctacttttt cagcgccaga ggcaggatcg accccctggt 1261 gtacctggac gagttcctga ggaacagcga gcccgtgctg aagagagtga acaggaccgg 1321 caacagcagc agcaacaagc aggagtacca gctgctgaag gacaacctgg tgcgcagcta 1381 caacaaggcc ctgaagaaga acgcccccta ccccatcttc gctatcaaga acggccctaa 1441 gagccacatc ggcaggcacc tgatgaccag ctttctgagc atgaagggcc tgaccgagct 1501 gacaaacgtg gtgggcaact ggagcgacaa gagggcctcc gccgtggcca ggaccaccta 1561 cacccaccag atcaccgcca tccccgacca ctacttcgcc ctggtgtcca ggtactacgc 1621 ctacgacccc atcagcaagg agatgatcgc cctgaaggac gagaccaacc ccatcgagga 1681 gtggcagcac atcgagcagc tgaagggcag cgccgagggc agcatcagat accccgcctg 1741 gaacggcatc atcagccagg aggtgctgga ctacctgagc agctacatca acaggcggat 1801 ctgagcggcc gcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc 1861 aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac 1921 atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca 1981 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat 2041 ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct 2101 gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc 2161 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt 2221 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt 2281 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc 2341 gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 2401 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 2461 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 2521 aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta 2581 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 2641 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga 2701 ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa 2761 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 2821 tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca 2881 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 2941 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 3001 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 3061 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 3121 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 3181 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 3241 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 3301 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 3361 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 3421 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 3481 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 3541 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 3601 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat 3661 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 3721 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt 3781 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta 3841 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 3901 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg 3961 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga 4021 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt 4081 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt 4141 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt 4201 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat 4261 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc 4321 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc 4381 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat 4441 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag 4501 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt 4561 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc 4621 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa 4681 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg 4741 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa 4801 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt cgacattgat 4861 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg 4921 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc 4981 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt 5041 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc 5101 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg 5161 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg 5221 ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct 5281 ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg 5341 gggggggg //