LOCUS Exported 7408 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses aa1-396 of Flp recombinase fused to ABI CID. ACCESSION . VERSION . KEYWORDS pBW2286_pCAG-PV1-FlpO-N396-L1-ABI-NLS-BGHpA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7408) 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 7408) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7408 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(22..38) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(22..38) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(35..57) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 46..62 /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(70..100) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 115..136 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin 221..355 /label=SV40 ori /note="SV40 origin of replication" primer_bind 282..301 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 375..509 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(412..431) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 466..485 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(577..594) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(748..1336) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(828..847) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1507..2367) /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 2130..2149 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2368..2472) /gene="bla" /label=AmpR promoter enhancer 2503..2882 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2884..3159 /label=chicken beta-actin promoter intron 3160..4175 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 4099..4121 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 4183..4202 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 6431..6454 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 6455..6475 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 6485..6709 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(6949..6968) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 7014..7069 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(7068..7087) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" ORIGIN 1 cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 61 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 121 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 181 gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc 241 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 301 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc 361 ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 421 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 481 tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg 541 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 601 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 661 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 721 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 781 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 841 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 901 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 961 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 1021 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 1081 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 1141 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg 1201 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 1261 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 1321 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 1381 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 1441 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 1501 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 1561 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 1621 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 1681 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 1741 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 1801 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 1861 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 1921 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 1981 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 2041 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 2101 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 2161 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 2221 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 2281 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 2341 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 2401 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 2461 aggggttccg cgcacatttc cccgaaaagt gccacctggg tcgacattga ttattgacta 2521 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 2581 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 2641 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 2701 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 2761 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 2821 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 2881 tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc 2941 caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg 3001 gggcgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggaaaggt 3061 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg 3121 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gttgccttcg 3181 ccccgtgccc cgctccgcgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta 3241 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt 3301 taatgacggc tcgtttcttt tctgtggctg cgtgaaagcc ttaaagggct ccgggagggc 3361 cctttgtgcg ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc 3421 cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg 3481 tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg 3541 gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg 3601 tgtgggcgcg gcggtcgggc tgtaaccccc ccctgcaccc ccctccccga gttgctgagc 3661 acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg 3721 gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg 3781 gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc 3841 gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa 3901 tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga 3961 agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc 4021 gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg 4081 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg 4141 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg 4201 tgctgtctca tcattttggc aaagaattga ttaattcgag cgaaggcgcg cccattactc 4261 gcatccattc tcaggctgtc tcgtctcgtc tcgcacgcgt gccgccacca tgagccagtt 4321 cgacatcctg tgcaagaccc cccccaaggt gctggtgcgg cagttcgtgg agagattcga 4381 gaggcccagc ggcgagaaga tcgccagctg tgccgccgag ctgacctacc tgtgctggat 4441 gatcacccac aacggcaccg ccatcaagag ggccaccttc atgagctaca acaccatcat 4501 cagcaacagc ctgagcttcg acatcgtgaa caagagcctg cagttcaagt acaagaccca 4561 gaaggccacc atcctggagg ccagcctgaa gaagctgatc cccgcctggg agttcaccat 4621 catcccttac aacggccaga agcaccagag cgacatcacc gacatcgtgt ccagcctgca 4681 gctgcagttc gagagcagcg aggaggccga caagggcaac agccacagca agaagatgct 4741 gaaggccctg ctgtccgagg gcgagagcat ctgggagatc accgagaaga tcctgaacag 4801 cttcgagtac accagcaggt tcaccaagac caagaccctg taccagttcc tgttcctggc 4861 cacattcatc aactgcggca ggttcagcga catcaagaac gtggacccca agagcttcaa 4921 gctggtgcag aacaagtacc tgggcgtgat cattcagtgc ctggtgaccg agaccaagac 4981 aagcgtgtcc aggcacatct actttttcag cgccagaggc aggatcgacc ccctggtgta 5041 cctggacgag ttcctgagga acagcgagcc cgtgctgaag agagtgaaca ggaccggcaa 5101 cagcagcagc aacaagcagg agtaccagct gctgaaggac aacctggtgc gcagctacaa 5161 caaggccctg aagaagaacg ccccctaccc catcttcgct atcaagaacg gccctaagag 5221 ccacatcggc aggcacctga tgaccagctt tctgagcatg aagggcctga ccgagctgac 5281 aaacgtggtg ggcaactgga gcgacaagag ggcctccgcc gtggccagga ccacctacac 5341 ccaccagatc accgccatcc ccgaccacta cttcgccctg gtgtccaggt actacgccta 5401 cgaccccatc agcaaggaga tgatcgccct gaaggacgag accaacccca tcgaggagtg 5461 gcagcacatc gagcagctga agggcagcgc cgagggctcc ggagggtctg gctccggatc 5521 aagtggtggc agcggtaccc ctttgtatgg ttttacttcg atttgtggaa gaagacctga 5581 gatggaagat gctgtttcga ctataccaag attccttcaa tcttcctctg gttcgatgtt 5641 agatggtcgg tttgatcctc aatccgccgc tcatttcttc ggtgtttacg acggccatgg 5701 cggttctcag gtagcgaact attgtagaga gaggatgcat ttggctttgg cggaggagat 5761 agctaaggag aaaccgatgc tctgcgatgg tgatacgtgg ctggagaagt ggaagaaagc 5821 tcttttcaac tcgttcctga gagttgactc ggagattggg tcagttgcgc cggagacggt 5881 tgggtcaacg tcggtggttg ccgttgtttt cccgtctcac atcttcgtcg ctaactgcgg 5941 tgactctaga gccgttcttt gccgcggcaa aactgcactt ccattatccg ttgaccataa 6001 accggataga gaagatgaag ctgcgaggat tgaagccgca ggagggaaag tgattcagtg 6061 gaatggagct cgtgttttcg gtgttctcgc catgtcgaga tccattggcg atagatactt 6121 gaaaccatcc atcattcctg atccggaagt gacggctgtg aagagagtaa aagaagatga 6181 ttgtctgatt ttggcgagtg acggggtttg ggatgtaatg acggatgaag aagcgtgtga 6241 gatggcaagg aagcggattc tcttgtggca caagaaaaac gcggtggctg gggatgcatc 6301 gttgctcgcg gatgagcgga gaaaggaagg gaaagatcct gcggcgatgt ccgcggctga 6361 gtatttgtca aagctggcga tacagagagg aagcaaagac aacataagtg tggtggtggt 6421 tgatttgaag gattacaagg acgatgacga taagcccaag aaaaagcgga aggtgtgaga 6481 attcctgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg 6541 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat 6601 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag 6661 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg 6721 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgt gtaccgctgg 6781 gagttcgtag acggaaacaa acgcagaatc caagcgcggc cgcccaggat acatagatta 6841 ccacaactcc gagcccttcc accgctagcg cactcctcag gtgcaggctg cctatcagaa 6901 ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc 6961 tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag 7021 gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac 7081 atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac 7141 atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat 7201 gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat 7261 tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca 7321 tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct 7381 tctcttatgg agatccctcg acctgcag //