LOCUS Exported 5941 bp ds-DNA circular SYN 26-SEP-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5941) AUTHORS Rui Lin TITLE Direct Submission JOURNAL Exported Sep 27, 2018 from SnapGene Viewer 4.2.5 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5941 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..141 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" promoter 159..473 /label=tight TRE promoter /note="Tet-responsive promoter PTight, consisting of seven tet operator sequences followed by the minimal CMV promoter" protein_bind 167..185 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 203..221 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 238..256 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 274..292 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 310..328 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 345..363 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 381..399 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 502..535 /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." protein_bind 586..619 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS complement(628..1926) /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" protein_bind complement(1939..1972) /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." protein_bind complement(2023..2056) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 2071..2679 /label=WPRE polyA_signal 2688..3164 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" repeat_region 3204..3344 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" rep_origin 3419..3874 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 4156..4260 /gene="bla" /label=AmpR promoter CDS 4261..5121 /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" rep_origin 5292..5880 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 121 actccatcac taggggttcc tgcggccgca cgcgtctcga gtttactccc tatcagtgat 181 agagaacgta tgtcgagttt actccctatc agtgatagag aacgatgtcg agtttactcc 241 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt 301 cgagtttact ccctatcagt gatagagaac gtatgtcgag tttatcccta tcagtgatag 361 agaacgtatg tcgagtttac tccctatcag tgatagagaa cgtatgtcga ggtaggcgtg 421 tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagaa 481 ttctctagag tcgactccgg aataacttcg tataggatac tttatacgaa gttatgcaga 541 atggtagctg gattgtagct gctattagca atatgaaacc tcttaataac ttcgtatagc 601 atacattata cgaagttatg gcgcgcctca gatccgcctg ttgatgtagc tgctcaggta 661 gtccagcacc tcctggctga tgatgccgtt ccaggcgggg tatctgatgc tgccctcggc 721 gctgcccttc agctgctcga tgtgctgcca ctcctcgatg gggttggtct cgtccttcag 781 ggcgatcatc tccttgctga tggggtcgta ggcgtagtac ctggacacca gggcgaagta 841 gtggtcgggg atggcggtga tctggtgggt gtaggtggtc ctggccacgg cggaggccct 901 cttgtcgctc cagttgccca ccacgtttgt cagctcggtc aggcccttca tgctcagaaa 961 gctggtcatc aggtgcctgc cgatgtggct cttagggccg ttcttgatag cgaagatggg 1021 gtagggggcg ttcttcttca gggccttgtt gtagctgcgc accaggttgt ccttcagcag 1081 ctggtactcc tgcttgttgc tgctgctgtt gccggtcctg ttcactctct tcagcacggg 1141 ctcgctgttc ctcaggaact cgtccaggta caccaggggg tcgatcctgc ctctggcgct 1201 gaaaaagtag atgtgcctgg acacgcttgt cttggtctcg gtcaccaggc actgaatgat 1261 cacgcccagg tacttgttct gcaccagctt gaagctcttg gggtccacgt tcttgatgtc 1321 gctgaacctg ccgcagttga tgaatgtggc caggaacagg aactggtaca gggtcttggt 1381 cttggtgaac ctgctggtgt actcgaagct gttcaggatc ttctcggtga tctcccagat 1441 gctctcgccc tcggacagca gggccttcag catcttcttg ctgtggctgt tgcccttgtc 1501 ggcctcctcg ctgctctcga actgcagctg caggctggac acgatgtcgg tgatgtcgct 1561 ctggtgcttc tggccgttgt aagggatgat ggtgaactcc caggcgggga tcagcttctt 1621 caggctggcc tccaggatgg tggccttctg ggtcttgtac ttgaactgca ggctcttgtt 1681 cacgatgtcg aagctcaggc tgttgctgat gatggtgttg tagctcatga aggtggccct 1741 cttgatggcg gtgccgttgt gggtgatcat ccagcacagg taggtcagct cggcggcaca 1801 gctggcgatc ttctcgccgc tgggcctctc gaatctctcc acgaactgcc gcaccagcac 1861 cttggggggg gtcttgcaca ggatgtcgaa ctggctcatc accttcctct tcttcttagg 1921 agccatggtg gcgctagcat aacttcgtat aaagtatcct atacgaagtt atttgcctta 1981 acccagaaat tatcactgtt attctttaga atggtgcaaa gaataacttc gtataatgta 2041 tgctatacga agttatgaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa 2101 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata 2161 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc 2221 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg 2281 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac 2341 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat 2401 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt 2461 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat 2521 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc 2581 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag 2641 tcggatctcc ctttgggccg cctccccgca tcgataccga gatctacggg tggcatccct 2701 gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc 2761 ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta 2821 tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct 2881 gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct 2941 ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg 3001 catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat 3061 tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat 3121 tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa 3181 ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct 3241 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc 3301 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta 3361 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac 3421 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct 3481 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg 3541 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt 3601 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca 3661 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga 3721 ctcttgttcc aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa 3781 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac 3841 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc 3901 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga 3961 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc 4021 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata 4081 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact 4141 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg 4201 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt 4261 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct 4321 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca 4381 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc 4441 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc 4501 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg 4561 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta 4621 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc 4681 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt 4741 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg 4801 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct 4861 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc 4921 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct 4981 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac 5041 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc 5101 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat 5161 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg 5221 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 5281 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 5341 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 5401 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta 5461 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 5521 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 5581 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 5641 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 5701 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 5761 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 5821 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 5881 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 5941 t //