LOCUS Exported 6185 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Gateway ORF clone of human TSC1 [NM_000368.4] without stop codon (for C-terminal fusions). ACCESSION . VERSION . KEYWORDS R777-E354 Hs.TSC1-nostop SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6185) TITLE Esposito lab REFERENCE 2 (bases 1 to 6185) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6185 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 23..45 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 37..54 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 38..54 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 84..183 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(3676..3775) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 4152..4943 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin 5036..5624 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5525..5544 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5778..5795 /label=L4440 /note="L4440 vector, forward primer" terminator 5954..5981 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 6073..6159 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" ORIGIN 1 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaaa 61 attcgagctc ggtccgagga tccgaaataa tgattttatt ttgactgata gtgacctgtt 121 cgttgcaaca aattgataag caatgctttt ttataatgcc aactttgtac aaaaaagttg 181 gcaccatggc ccaacaagca aatgtcgggg agcttcttgc catgctggac tcccccatgc 241 tgggtgtgcg ggacgacgtg acagctgtct ttaaagagaa cctcaattct gaccgtggcc 301 ctatgcttgt aaacaccttg gtggattatt acctggaaac cagctctcag ccggcattgc 361 acatcctgac caccttgcaa gagccacatg acaagcacct cttggacagg attaacgaat 421 atgtgggcaa agccgccact cgtttatcca tcctctcgtt actgggtcat gtcataagac 481 tgcagccatc ttggaagcat aagctctctc aagcacctct tttgccttct ttactaaaat 541 gtctcaagat ggacactgac gtcgttgtcc tcacaacagg cgtcttggtg ttgataacca 601 tgctaccaat gattccacag tctgggaaac agcatcttct tgatttcttt gacatttttg 661 gccgtctgtc atcatggtgc ctgaagaaac caggccacgt ggcggaagtc tatctcgtcc 721 atctccatgc cagtgtgtac gcactctttc atcgccttta tggaatgtac ccttgcaact 781 tcgtctcctt tttgcgttct cattacagta tgaaagaaaa cctggagact tttgaagaag 841 tggtcaagcc aatgatggag catgtgcgaa ttcatccgga attagtgact ggatccaagg 901 accatgaact ggaccctcga aggtggaaga gattagaaac tcatgatgtt gtgatcgagt 961 gtgccaaaat ctctctggat cccacagaag cctcatatga agatggctat tctgtgtctc 1021 accaaatctc agcccgcttt cctcatcgtt cagccgatgt caccaccagc ccttatgctg 1081 acacacagaa tagctatggg tgtgctactt ctacccctta ctccacgtct cggctgatgt 1141 tgttaaatat gccagggcag ctacctcaga ctctgagttc cccatcgaca cggctgataa 1201 ctgaaccacc acaagctact ctttggagcc catctatggt ttgtggtatg accactcctc 1261 caacttctcc tggaaatgtc ccacctgatc tgtcacaccc ttacagtaaa gtctttggta 1321 caactgcagg tggaaaagga actcctctgg gaaccccagc aacctctcct cctccagccc 1381 cactctgtca ttcggatgac tacgtgcaca tttcactccc ccaggccaca gtcacacccc 1441 ccaggaagga agagagaatg gattctgcaa gaccatgtct acacagacaa caccatcttc 1501 tgaatgacag aggatcagaa gagccacctg gcagcaaagg ttctgtcact ctaagtgatc 1561 ttccagggtt tttaggtgat ctggcctctg aagaagatag tattgaaaaa gataaagaag 1621 aagctgcaat atctagagaa ctttctgaga tcaccacagc agaggcagag cctgtggttc 1681 ctcgaggagg ctttgactct cccttttacc gagacagtct cccaggttct cagcggaaga 1741 cccactcggc agcctccagt tctcagggcg ccagcgtgaa ccctgagcct ttacactcct 1801 ccctggacaa gcttgggcct gacacaccaa agcaagcctt tactcccata gacctgccct 1861 gcggcagtgc tgatgaaagc cctgcgggag acagggaatg ccagacttct ttggagacca 1921 gtatcttcac tcccagtcct tgtaaaattc cacctccgac gagagtgggc tttggaagcg 1981 ggcagcctcc cccgtatgat catctttttg aggtggcatt gccaaagaca gcccatcatt 2041 ttgtcatcag gaagactgag gagctgttaa agaaagcaaa aggaaacaca gaggaagatg 2101 gtgtgccctc tacctcccca atggaagtgc tggacagact gatacagcag ggagcagacg 2161 cgcacagcaa ggagctgaac aagttgcctt tacccagcaa gtctgtcgac tggacccact 2221 ttggaggctc tcctccttca gatgagatcc gcaccctccg agaccagttg cttttactgc 2281 acaaccagtt actctatgag cgttttaaga ggcagcagca tgccctccgg aacaggcggc 2341 tcctccgcaa ggtgatcaaa gcagcagctc tggaggaaca taatgctgcc atgaaagatc 2401 agttgaagtt acaagagaag gacatccaga tgtggaaggt tagtctgcag aaagaacaag 2461 ctagatacaa tcagctccag gagcagcgtg acactatggt aaccaagctc cacagccaga 2521 tcagacagct gcagcatgac cgagaggaat tctacaacca gagccaggaa ttacagacga 2581 agctggagga ctgcaggaac atgattgcgg agctgcggat agaactgaag aaggccaaca 2641 acaaggtgtg tcacactgag ctgctgctca gtcaggtttc ccaaaagctc tcaaacagtg 2701 agtcggtcca gcagcagatg gagttcttga acaggcagct gttggttctt ggggaggtca 2761 acgagctcta tttggaacaa ctgcagaaca agcactcaga taccacaaag gaagtagaaa 2821 tgatgaaagc cgcctatcgg aaagagctag aaaaaaacag aagccatgtt ctccagcaga 2881 ctcagaggct tgatacctcc caaaaacgga ttttggaact ggaatctcac ctggccaaga 2941 aagaccacct tcttttggaa cagaagaaat atctagagga tgtcaaactc caggcaagag 3001 gacagctgca ggccgcagag agcaggtatg aggctcagaa aaggataacc caggtgtttg 3061 aattggagat cttagattta tatggcaggt tggagaaaga tggcctcctg aaaaaacttg 3121 aagaagaaaa agcagaagca gctgaagcag cagaagaaag gcttgactgt tgtaatgacg 3181 ggtgctcaga ttccatggta gggcacaatg aagaggcatc tggccacaac ggtgagacca 3241 agacccccag gcccagcagc gcccggggca gtagtggaag cagaggtggt ggaggcagca 3301 gcagcagcag cagcgagctt tctaccccag agaaaccccc acaccagagg gcaggcccat 3361 tcagcagtcg gtgggagacg actatgggag aagcgtctgc cagcatcccc accactgtgg 3421 gctcacttcc cagttcaaaa agcttcctgg gtatgaaggc tcgagagtta tttcgtaata 3481 agagcgagag ccagtgtgat gaggacggca tgaccagtag cctttctgag agcctaaaga 3541 cagaactggg caaagacttg ggtgtggaag ccaagattcc cctgaaccta gatggccctc 3601 acccgtctcc cccgaccccg gacagtgttg gacagctaca tatcatggac tacaatgaga 3661 ctcatcatga acacagctca actttcttgt acaaagttgg cattatagga aagcattgct 3721 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgcatgc 3781 aagcttagat cctctagacc agccaggaca gaaatgcctc gacttcgctg ctacccaagg 3841 ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg aacccagtgg acataagcct 3901 gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct 3961 tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt 4021 tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc 4081 gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa 4141 agttaaacat tatgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag 4201 ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg 4261 cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg gtgaccgtaa 4321 ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc 4381 ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca 4441 ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca 4501 ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa 4561 aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg 4621 ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc 4681 ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg 4741 cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc 4801 cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag 4861 atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg aaaggcgaga 4921 tcaccaaggt agtcggcaaa taaccctcga gccacccatg accaaaatcc cttaacgtga 4981 gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag 5041 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 5101 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 5161 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga 5221 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 5281 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 5341 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 5401 ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa 5461 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 5521 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 5581 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg 5641 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat 5701 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca 5761 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca 5821 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg 5881 actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc caggaagagt 5941 ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt gatgcctggc 6001 agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc 6061 cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg 6121 aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag ttccctactc 6181 tcgcg //