LOCUS Exported 3422 bp DNA circular SYN 24-JUL-2020 DEFINITION synthetic linear DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2001) AUTHORS Keoni Gandall TITLE Direct Submission JOURNAL Exported from Poly. FEATURES Location/Qualifiers terminator 1..3422 /note="transcription terminator for bacteriophage T7 RNA polymerase" /label="T7 terminator" terminator 38..85 /label="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind 1933..1949 /label="M13 for" /note="common sequencing primer, one of multiple similar variants" terminator 1950..1981 /label="tonB terminator" /note="bidirectional E. coli tonB-P14 transcription terminator" CDS 2030..3328 /label="tilS" /note="Ligates lysine onto the cytidine present at position 34 of the AUA codon-specific tRNA(Ile) that contains the anticodon CAU, in an ATP-dependent manner. Cytidine is converted to lysidine, thus changing the amino acid specificity of the tRNA from methionine to isoleucine. This enzyme is essential for viability." /translation="MTLTLNRQLLTSRQILVAFSGGLDSTVLLHQLVQWRTENPGVALRAIHVHHGLSANADAWVTHCENVCQQWQVPLVVERVQLAQEGLGIEAQARQARYQAFARTLLPGEVLVTAQHLDDQCETFLLALKRGSGPAGLSAMAEVSEFAGTRLIRPLLARTRGELVQWARQYDLRWIEDESNQDDSYDRNFLRLRVVPLLQQRWPHFAEATARSAALCAEQESLLDELLADDLAHCQSPQGTLQIVPMLAMSDARRAAIIRRWLAGQNAPMPSRDALVRIWQEVALAREDASPCLRLGAFEIRRYQSQLWWIKSVTGQSENIVPWQTWLQPLELPAGLGSVQLNAGGDIRPPRADEAVSVRFKAPGLLHIVGRNGGRKLKKIWQELGVPPWLRDTTPLLFYGETLIAAAGVFVTQEGVAEGENGVSFVWQKTLS" primer_bind complement(90..106) /label="M13 rev" /note="common sequencing primer, one of multiple similar variants" rep_origin complement(173..761) /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(952..1812) /label="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*" promoter complement(1813..1917) /note="ampicillin resistance gene promoter" /label="bla promoter" misc_feature 3363..3422 /note="FreeGenes Barcode. Defined as I-SceI site flanked by 21 base pair random sequences." /label="FreeGenes Barcode" ORIGIN 1 CGCTGAGGTG TCAATCGTCG GAGCCGCTGA GCAATAACTA GCATAACCCC TTGGGGCCTC 61 TAAACGGGTC TTGAGGGGTT TTTTGCATGG TCATAGCTGT TTCCTGAGAG CTTGGCAGGT 121 GATGACACAC ATTAACAAAT TTCGTGAGGA GTCTCCAGAA GAATGCCATT AATTTCCATA 181 GGCTCCGCCC CCCTGACGAG CATCACAAAA ATCGACGCTC AAGTCAGAGG TGGCGAAACC 241 CGACAGGACT ATAAAGATAC CAGGCGTTTC CCCCTGGAAG CTCCCTCGTG CGCTCTCCTG 301 TTCCGACCCT GCCGCTTACC GGATACCTGT CCGCCTTTCT CCCTTCGGGA AGCGTGGCGC 361 TTTCTCATAG CTCACGCTGT AGGTATCTCA GTTCGGTGTA GGTCGTTCGC TCCAAGCTGG 421 GCTGTGTGCA CGAACCCCCC GTTCAGCCCG ACCGCTGCGC CTTATCCGGT AACTATCGTC 481 TTGAGTCCAA CCCGGTAAGA CACGACTTAT CGCCACTGGC AGCAGCCACT GGTAACAGGA 541 TTAGCAGAGC GAGGTATGTA GGCGGTGCTA CAGAGTTCTT GAAGTGGTGG CCTAACTACG 601 GCTACACTAG AAGAACAGTA TTTGGTATCT GCGCTCTGCT GAAGCCAGTT ACCTTCGGAA 661 AAAGAGTTGG TAGCTCTTGA TCCGGCAAAC AAACCACCGC TGGTAGCGGT GGTTTTTTTG 721 TTTGCAAGCA GCAGATTACG CGCAGAAAAA AAGGATCTCA AGAAGGCCTA CTATTAGCAA 781 CAACGATCCT TTGATCTTTT CTACGGGGTC TGACGCTCAG TGGAACGAAA ACTCACGTTA 841 AGGGATTTTG GTCATGAGAT TATCAAAAAG GATCTTCACC TAGATCCTTT TAAATTAAAA 901 ATGAAGTTTT AAATCAATCT AAAGTATATA TGAGTAAACT TGGTCTGACA GTTACCAATG 961 CTTAATCAGT GAGGCACCTA TCTCAGCGAT CTGTCTATTT CGTTCATCCA TAGTTGCCTG 1021 ACTCCCCGTC GTGTAGATAA CTACGATACG GGAGGGCTTA CCATCTGGCC CCAGTGCTGC 1081 AATGATACCG CGAGAACCAC GCTCACCGGC TCCAGATTTA TCAGCAATAA ACCAGCCAGC 1141 CGGAAGGGCC GAGCGCAGAA GTGGTCCTGC AACTTTATCC GCCTCCATCC AGTCTATTAA 1201 TTGTTGCCGG GAAGCTAGAG TAAGTAGTTC GCCAGTTAAT AGTTTGCGCA ACGTTGTTGC 1261 CATTGCTACA GGCATCGTGG TGTCACGCTC GTCGTTTGGT ATGGCTTCAT TCAGCTCCGG 1321 TTCCCAACGA TCAAGGCGAG TTACATGATC CCCCATGTTG TGCAAAAAAG CGGTTAGCTC 1381 CTTCGGTCCT CCGATCGTTG TCAGAAGTAA GTTGGCCGCA GTGTTATCAC TCATGGTTAT 1441 GGCAGCACTG CATAATTCTC TTACTGTCAT GCCATCCGTA AGATGCTTTT CTGTGACTGG 1501 TGAGTACTCA ACCAAGTCAT TCTGAGAATA GTGTATGCGG CGACCGAGTT GCTCTTGCCC 1561 GGCGTCAATA CGGGATAATA CCGCGCCACA TAGCAGAACT TTAAAAGTGC TCATCATTGG 1621 AAAACGTTCT TCGGGGCGAA AACTCTCAAG GATCTTACCG CTGTTGAGAT CCAGTTCGAT 1681 GTAACCCACT CGTGCACCCA ACTGATCTTC AGCATCTTTT ACTTTCACCA GCGTTTCTGG 1741 GTGAGCAAAA ACAGGAAGGC AAAATGCCGC AAAAAAGGGA ATAAGGGCGA CACGGAAATG 1801 TTGAATACTC ATACTCTTCC TTTTTCAATA TTATTGAAGC ATTTATCAGG GTTATTGTCT 1861 CATGAGCGGA TACATATTTG AATGTATTTA GAAAAATAAA CAAATAGGGG TTCCGCGCAC 1921 CTGCACCAGT CAGTAAAACG ACGGCCAGTA GTCAAAAGCC TCCGACCGGA GGCTTTTGAC 1981 TTGGTTCAGG TGGAGTGGGA GAAACCGGAG CCATACAGTA CGGTCTCTAA TGACCCTAAC 2041 CCTAAACCGT CAGCTACTAA CCTCACGTCA GATCCTAGTC GCATTCTCAG GAGGACTAGA 2101 CTCAACCGTC CTACTACACC AGCTAGTCCA GTGGCGTACC GAAAACCCGG GAGTCGCACT 2161 ACGTGCAATC CACGTCCACC ACGGACTATC AGCAAACGCA GACGCATGGG TCACCCACTG 2221 CGAAAACGTC TGCCAGCAGT GGCAGGTCCC GCTAGTCGTC GAACGTGTCC AGCTAGCACA 2281 GGAAGGACTA GGAATCGAAG CACAGGCACG TCAGGCACGT TACCAGGCAT TCGCACGTAC 2341 CCTACTACCG GGAGAAGTCC TAGTCACCGC ACAGCACCTA GACGACCAGT GCGAAACCTT 2401 CCTACTAGCA CTAAAACGTG GATCAGGACC GGCAGGACTA TCAGCAATGG CAGAAGTCTC 2461 AGAATTCGCA GGAACCCGTC TAATCCGTCC GCTACTAGCA CGTACCCGTG GAGAACTAGT 2521 CCAGTGGGCA CGTCAGTACG ACCTACGTTG GATCGAAGAC GAATCAAACC AGGACGACTC 2581 ATACGACCGT AACTTCCTAC GTCTACGTGT CGTCCCGCTA CTACAGCAGC GTTGGCCGCA 2641 CTTCGCAGAA GCAACCGCAC GTTCAGCAGC ACTATGCGCA GAACAGGAAT CACTACTAGA 2701 CGAACTACTA GCAGACGACC TAGCACACTG CCAGTCACCG CAGGGAACCC TACAGATCGT 2761 CCCGATGCTA GCAATGTCAG ACGCACGTCG TGCAGCAATC ATCCGTCGTT GGCTAGCAGG 2821 ACAGAACGCA CCGATGCCGT CACGTGACGC ACTAGTCCGT ATCTGGCAGG AAGTCGCACT 2881 AGCACGTGAA GACGCATCAC CGTGCCTACG TCTAGGAGCA TTCGAAATCC GTCGTTACCA 2941 GTCACAGCTA TGGTGGATCA AATCAGTCAC CGGACAGTCA GAAAACATCG TCCCGTGGCA 3001 GACCTGGCTA CAGCCGCTAG AACTACCGGC AGGACTAGGA TCAGTCCAGC TAAACGCAGG 3061 AGGAGACATC CGTCCGCCGC GTGCAGACGA AGCAGTCTCA GTCCGTTTCA AAGCACCGGG 3121 ACTACTACAC ATCGTCGGAC GTAACGGAGG ACGTAAACTA AAAAAAATCT GGCAGGAACT 3181 AGGAGTCCCG CCGTGGCTAC GTGACACCAC CCCGCTACTA TTCTACGGAG AAACCCTAAT 3241 CGCAGCAGCA GGAGTCTTCG TCACCCAGGA AGGAGTCGCA GAAGGAGAAA ACGGAGTCTC 3301 ATTCGTCTGG CAGAAAACCC TATCATGAAG AGCTTCGAGA CCAGATAGTC ACGCACAAGA 3361 CCATGCGTGC TTACCCGCGC ATATAGGGAT AACAGGGTAA TTATTTGGTC ACCGCTCGAA 3421 AT //