LOCUS Exported 2975 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..2975 /label="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" 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..2881 /label="tsf" /note="(Microbial infection) Promotes the tRNase activity of CdiA-CT from E.coli strain NC101 (CdiA-CT-NC101); required in vivo and in vitro. Probably loads charged tRNA onto EF-Tu, making more ternary GTP-EF-Tu-aa-tRNA complexes. The guanine nucleotide exchange factor capacity of this protein does not seem to be needed as no GTP hydrolysis occurs during tRNA cleavage. CdiA-CT is the toxic component of a toxin-immunity protein module, which functions as a cellular contact-dependent growth inhibition (CDI) system. CDI modules allow bacteria to communicate with and inhibit the growth of closely related neighboring bacteria in a contact-dependent fashion (PubMed:28973472). EF-Ts interacts with at least 2 different toxic CT domains, the 2 toxins are different and degrade tRNA at different positions (PubMed:28973472, PubMed:28223500)." /translation="MAEITASLVKELRERTGAGMMDCKKALTEANGDIELAIENMRKSGAIKAAKKAGNVAADGVIKTKIDGNYGIILEVNCQTDFVAKDAGFQAFADKVLDAAVAGKITDVEVLKAQFEEERVALVAKIGENINIRRVAALEGDVLGSYQHGARIGVLVAAKGADEELVKHIAMHVAASKPEFIKPEDVSAEVVEKEYQVQLDIAMQSGKPKEIAEKMVEGRMKKFTGEVSLTGQPFVMEPSKTVGQLLKEHNAEVTGFIRFEVGEGIEKVETDFAAEVAAMSKQS" 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) /label="bla promoter" /note="ampicillin resistance gene promoter" misc_feature 2916..2975 /label="FreeGenes Barcode" /note="FreeGenes Barcode. Defined as I-SceI site flanked by 21 base pair random sequences." 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 GAAAGCACTC TTAGGCCTCT GGGTCTCTAA TGGCAGAAAT 2041 CACCGCATCA CTAGTCAAAG AACTACGTGA ACGTACCGGA GCAGGAATGA TGGACTGCAA 2101 AAAAGCACTA ACCGAAGCAA ACGGAGACAT CGAACTAGCA ATCGAAAACA TGCGTAAATC 2161 AGGAGCAATC AAAGCAGCAA AAAAAGCAGG AAACGTCGCA GCAGACGGAG TCATCAAAAC 2221 CAAAATCGAC GGAAACTACG GAATCATCCT AGAAGTCAAC TGCCAGACCG ACTTCGTCGC 2281 AAAAGACGCA GGATTCCAGG CATTCGCAGA CAAAGTCCTA GACGCAGCAG TCGCAGGAAA 2341 AATCACCGAC GTCGAAGTCC TAAAAGCACA GTTCGAAGAA GAACGTGTCG CACTAGTCGC 2401 AAAAATCGGA GAAAACATCA ACATCCGTCG TGTCGCAGCA CTAGAAGGAG ACGTCCTAGG 2461 ATCATACCAG CACGGAGCAC GTATCGGAGT CCTAGTCGCA GCAAAAGGAG CAGACGAAGA 2521 ACTAGTCAAA CACATCGCAA TGCACGTCGC AGCATCAAAA CCGGAATTCA TCAAACCGGA 2581 AGACGTCTCA GCAGAAGTCG TCGAAAAAGA ATACCAGGTC CAGCTAGACA TCGCAATGCA 2641 GTCAGGAAAA CCGAAAGAAA TCGCAGAAAA AATGGTCGAA GGACGTATGA AAAAATTCAC 2701 CGGAGAAGTC TCACTAACCG GACAGCCGTT CGTCATGGAA CCGTCAAAAA CCGTCGGACA 2761 GCTACTAAAA GAACACAACG CAGAAGTCAC CGGATTCATC CGTTTCGAAG TCGGAGAAGG 2821 AATCGAAAAA GTCGAAACCG ACTTCGCAGC AGAAGTCGCA GCAATGTCAA AACAGTCATG 2881 AAGAGCTTCG AGACCATTAG TACACTCCGT GAGCGAATCA ATAAGCTATA CCTAGCTAGG 2941 GATAACAGGG TAATATCGGG CTAACGAAGA TTGGT //