LOCUS Exported 5897 bp ds-DNA circular SYN 15-DEC-2017 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM recombinant plasmid REFERENCE 1 (bases 1 to 5897) AUTHORS Addgene TITLE Direct Submission JOURNAL Exported Dec 15, 2017 from SnapGene 4.1.2 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5897 /organism="recombinant plasmid" /mol_type="other DNA" protein_bind 106..127 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 142..172 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 180..196 /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 204..220 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" misc_feature 245..385 /label=ITR WT (141bp) repeat_region 245..385 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" misc_feature 483..1299 /label=CBh /note="CBh" misc_feature 1309..1341 /label=MCS CDS 1342..2040 /codon_start=1 /product="monomeric far-red fluorescent protein (Shcherbo et al., 2009)" /label=mKate2 /note="mammalian codon-optimized" /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY CDLPSKLGHR" misc_feature 2069..2621 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" misc_feature 2634..2729 /label=MCS 2 enhancer 2731..3339 /gene="WPRE" polyA_signal 3356..3477 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" misc_feature 3515..3655 /label=ITR WT (141bp) repeat_region 3515..3655 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" promoter 3991..4095 /gene="bla" /label=AmpR promoter CDS 4096..4956 /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 5127..5715 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct 121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat 181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc 241 atgccctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg 301 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg 361 gccaactcca tcactagggg ttcctgcggc cgcagccgtg ttttagagct agaaatagca 421 agttaaaata aggctagtcc gtagcgcgtg cgccaattct gcagacaaat tctagaggta 481 cccgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca 541 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt 601 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg 661 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag 721 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt 781 accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca 841 cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg 901 gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg 961 agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg 1021 cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgacg 1081 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact 1141 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta 1201 gctgagcaag aggtaagggt ttaagggatg gttggttggt ggggtattaa tgtttaatta 1261 cctggagcac ctgcctgaaa tcactttttt tcaggttgga ccggtgccgt ttaaacaaca 1321 attggaagag ctcgtcgacc catggtgagc gagctgatta aggagaacat gcacatgaag 1381 ctgtacatgg agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc 1441 aagccctacg agggcaccca gaccatgaga atcaaggcgg tcgagggcgg ccctctcccc 1501 ttcgccttcg acatcctggc taccagcttc atgtacggca gcaaaacctt catcaaccac 1561 acccagggca tccccgactt ctttaagcag tccttccccg agggcttcac atgggagaga 1621 gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac 1681 ggctgcctca tctacaacgt caagatcaga ggggtgaact tcccatccaa cggccctgtg 1741 atgcagaaga aaacactcgg ctgggaggcc agcaccgaga ccctgtaccc cgctgacggc 1801 ggcctggaag gcagagccga catggccctg aagctcgtgg gcgggggcca cctgatctgc 1861 aacttgaaga ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc 1921 tactatgtgg acagaagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag 1981 cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg gcacagatga 2041 gaattcgatt acggcggccc cccccctaac gttactggcc gaagccgctt ggaataaggc 2101 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg 2161 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc 2221 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga 2281 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg 2341 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag 2401 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc 2461 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct 2521 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac 2581 cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatgctggc 2641 cggcctactt aattaaccac tagttctgga tccttaaaac tcgaggccta taacgcgtat 2701 agctagcttt gcgatcgcaa aggcgcgcca gcttatcgat aatcaacctc tggattacaa 2761 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata 2821 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc 2881 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg 2941 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac 3001 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat 3061 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt 3121 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat 3181 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc 3241 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag 3301 tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctggaga gatcttaaga 3361 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt 3421 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttctg 3481 attttgtagt taacacgtgc gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc 3541 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc 3601 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc 3661 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact 3721 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc 3781 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc 3841 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga 3901 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag 3961 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa 4021 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat 4081 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg 4141 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa 4201 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt 4261 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt 4321 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat 4381 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg 4441 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta 4501 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat 4561 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag 4621 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa 4681 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca 4741 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc 4801 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt 4861 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc 4921 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat 4981 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt 5041 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac 5101 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 5161 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 5221 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta 5281 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 5341 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 5401 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 5461 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta 5521 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 5581 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 5641 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 5701 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 5761 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc 5821 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg 5881 agcgaggaag cggaaga //