LOCUS Exported 6195 bp ds-DNA circular SYN 10-JUN-2017 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pAAV-TIWB-myrmScarlet-I-P2A-kSPSH3S11Nlg(d4)+sequencing result SOURCE synthetic DNA construct ORGANISM recombinant plasmid REFERENCE 1 (bases 1 to 6195) AUTHORS Lab of Neurobiology TITLE Direct Submission JOURNAL Exported Wednesday, May 2, 2018 from SnapGene Viewer 3.3.3 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6195 /organism="recombinant plasmid" /mol_type="other DNA" repeat_region 1..141 /note="AAV2 ITR" /note="inverted terminal repeat of adeno-associated virus serotype 2" protein_bind 165..183 /gene="tetO" /bound_moiety="tetracycline repressor TetR" /note="tet operator" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 201..219 /gene="tetO" /bound_moiety="tetracycline repressor TetR" /note="tet operator" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 237..255 /gene="tetO" /bound_moiety="tetracycline repressor TetR" /note="tet operator" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 273..291 /gene="tetO" /bound_moiety="tetracycline repressor TetR" /note="tet operator" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 309..327 /gene="tetO" /bound_moiety="tetracycline repressor TetR" /note="tet operator" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 345..363 /gene="tetO" /bound_moiety="tetracycline repressor TetR" /note="tet operator" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 381..399 /gene="tetO" /bound_moiety="tetracycline repressor TetR" /note="tet operator" /note="bacterial operator O2 for the tetR and tetA genes" CDS 405..475 /note="Modified Minimal CMV promoter" intron 583..715 /note="chimeric intron" /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" promoter 760..778 /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" CDS 839..1530 /codon_start=1 /product="bright monomeric red fluorescent protein evolved from a synthetic template (Bindels et al., 2016)" /note="mScarlet-I" /note="matures faster than mScarlet" /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGGA VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS TGGMDELY" CDS 1541..1597 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /note="P2A" /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 1886..1933 /codon_start=1 /product="11th beta-strand of superfolder GFP" /note="GFP11" /note="associates with GFP(1-10), consisting of the first 10 beta-strands, to reconstitute fluorescent GFP (Cabantous et al., 2005; Kamiyama et al., 2016)" /translation="RDHMVLHEYVNAAGIT" CDS complement(2228..2239) /codon_start=1 /product="Factor Xa recognition and cleavage site" /note="Factor Xa site" /translation="IEGR" misc_feature 2625..3213 /note="WPRE" /note="woodchuck hepatitis virus posttranscriptional regulatory element" CDS complement(3096..3107) /codon_start=1 /product="Factor Xa recognition and cleavage site" /note="Factor Xa site" /translation="IEGR" polyA_signal 3226..3450 /note="bGH poly(A) signal" /note="bovine growth hormone polyadenylation signal" repeat_region 3458..3598 /note="AAV2 ITR" /note="inverted terminal repeat of adeno-associated virus serotype 2" rep_origin 3673..4128 /direction=RIGHT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 4410..4514 /gene="bla" /note="AmpR promoter" CDS 4515..5375 /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 5546..6134 /direction=RIGHT /note="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 cgcgttggct ttactcccta tcagtgatag 181 agaacgtatg aagagtttac tccctatcag tgatagagaa cgtatgcaga ctttactccc 241 tatcagtgat agagaacgta taaggagttt actccctatc agtgatagag aacgtatgac 301 cagtttactc cctatcagtg atagagaacg tatctacagt ttactcccta tcagtgatag 361 agaacgtata tccagtttac tccctatcag tgatagagaa cgtataagct ttaggcgtgt 421 acggtgggcg cctataaaag cagagctcgt ttagtgaacc gtcagatcgc ctggagcaat 481 tccacaacac ttttgtctta taccaacttt ccgtaccact tcctaccctc gtaaagctag 541 tcgaacttaa gctgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca 601 agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt 661 ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc 721 cactcccagt tcaattacag ctcttaaggc tagagtactt aatacgactc actataggct 781 agccaccatg ggctgtgtgc agtgtaagga caaggaggct accaagctga caggaggcgt 841 gagcaagggc gaggcagtga tcaaggagtt catgcggttc aaggtgcaca tggagggctc 901 catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac 961 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttctcct gggacatcct 1021 gtcccctcag ttcatgtacg gctccagggc cttcatcaag caccccgccg acatccccga 1081 ctactataag cagtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga 1141 cggcggcgcc gtgaccgtga cccaggacac ctccctggag gacggcaccc tgatctacaa 1201 ggtgaagctc cgcggcacca acttccctcc tgacggcccc gtaatgcaga agaagacaat 1261 gggctgggaa gcgtccaccg agcggttgta ccccgaggac ggcgtgctga agggcgacat 1321 taagatggcc ctgcgcctga aggacggcgg ccgctacctg gcggacttca agaccaccta 1381 caaggccaag aagcccgtgc agatgcccgg cgcctacaac gtcgaccgca agttggacat 1441 cacctcccac aacgaggact acaccgtggt ggaacagtac gaacgctccg agggccgcca 1501 ctccaccggc ggcatggacg agctgtacaa gggaagcgga gctactaact tcagcctgct 1561 gaagcaggct ggagacgtgg aggagaaccc tggacctgag accgacaccc tcctgctatg 1621 ggtgctgctg ctctgggtgc ccggcagcac cggcgacgca ccggtcggag gcaacgaccc 1681 taacctgttc gtcgccctgt acgacttcgt cgcctctggc gacaacaccc tgagcatcac 1741 caagggcgag aagctgagag tcctgggcta caaccacaac ggcgagtggt gcgaggccca 1801 gaccaagaac ggccagggct gggtccctag caactacatc acacctgtca acagcaccgg 1861 tggaggcagc ggaggaggct ccggaagaga ccacatggtg ctgcacgagt acgtgaacgc 1921 tgctggaatc acaggaggag gctccggagg cggaagcggt accctcgagc tggtgcccca 1981 cttgcataac ctcaatgaca tcagccaata cacaagtaca accaccaagg tcccaagcac 2041 tgacatcacc cttcgaccaa ctcggaaaaa ctcaacccct gtgaccagcg catttccaac 2101 cgctaagcaa gacgacccca aacaacagcc tagtccattt agcgtagacc aaagggacta 2161 cagcacagag ctgtccgtga ccatagctgt aggagcttcc ctgcttttcc tgaacattct 2221 ggctttcgcg gccctctatt acaagaagga caaaaggcgc cacgacgtac ataggagatg 2281 tagtccgcaa cgaactacaa ccaatgatct tacgcacgca cctgaggagg agatcatgag 2341 tcttcagatg aaacacacag acctggacca cgagtgtgaa tccatccatc cccatgaggt 2401 ggtgctgagg acagcatgcc ctccggacta taccctggca atgcgccgaa gcccagacga 2461 cattcccctg atgacaccca ataccataac catgattccc aacactattc caggaatcca 2521 gcctctccac actttcaaca cattcactgg tggacagaat aatactctcc ctcatccgca 2581 tccacacccc cactcccaca gctaagaatt cctgcagccc cgataatcaa cctctggatt 2641 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg 2701 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct 2761 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc 2821 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca 2881 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac 2941 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt 3001 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct 3061 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc 3121 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga 3181 cgagtcggat ctccctttgg gccgcctccc cgcatcgggg ggatcctgtg ccttctagtt 3241 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc 3301 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt 3361 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca 3421 ggcatgctgg ggatgcggtg ggctctatgg cggccgcagg aacccctagt gatggagttg 3481 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga 3541 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg 3601 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa 3661 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc 3721 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt 3781 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag 3841 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt 3901 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt 3961 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt 4021 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt 4081 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc 4141 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg 4201 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg 4261 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa 4321 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga 4381 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa 4441 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 4501 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg 4561 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag 4621 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg 4681 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg 4741 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt 4801 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga 4861 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac 4921 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc 4981 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc 5041 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac 5101 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag 5161 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg 5221 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta 5281 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg 5341 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata 5401 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt 5461 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 5521 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 5581 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 5641 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag 5701 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 5761 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 5821 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 5881 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 5941 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 6001 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 6061 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 6121 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc 6181 cttttgctca catgt //