LOCUS Exported 3455 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..3455 /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 100..3454 /label="secY" /note="The central subunit of the protein translocation channel SecYEG. Consists of two halves formed by TMs 1-5 and 6-10. These two domains form a lateral gate at the front which open onto the bilayer between TMs 2 and 7, and are clamped together by SecE at the back. The channel is closed by both a pore ring composed of hydrophobic SecY resides and a short helix (helix 2A) on the extracellular side of the membrane which forms a plug. The plug probably moves laterally to allow the channel to open. The ring and the pore may move independently. SecY is required to insert newly synthesized SecY into the inner membrane. Overexpression of some hybrid proteins has been thought to jam the protein secretion apparatus resulting in cell death; while this may be true, overexpression also results in FtsH-mediated degradation of SecY." /translation="MAKQPGLDFQSAKGGLGELKRRLLFVIGALIVFRIGSFIPIPGIDAAVLAKLLEQQRGTIIEMFNMFSGGALSRASIFALGIMPYISASIIIQLLTVVHPTLAEIKKEGESGRRKISQYTRYGTLVLAIFQSIGIATGLPNMPGMQGLVINPGFAFYFTAVVSLVTGTMFLMWLGEQITERGIGNGISIIIFAGIVAGLPPAIAHTIEQARQGDLHFLVLLLVAVLVFAVTFFVVFVERGQRRIVVNYAKRQQGRRVYAAQSTHLPLKVNMAGVIPAIFASSIILFPATIASWFGGGTGWNWLTTISLYLQPGQPLYVLLYASAIIFFCFFYTALVFNPRETADNLKKSGAFVPGIRPGEQTAKYIDKVMTRLTLVGALYITFICLIPEFMRDAMKVPFYFGGTSLLIVVVVIMDFMAQVQTLMMSSQYESALKKANLKGYGR" 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 3396..3455 /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 TGGCAAAACA 2041 GCCGGGACTA GACTTCCAGT CAGCAAAAGG AGGACTAGGA GAACTAAAAC GTCGTCTACT 2101 ATTCGTCATC GGAGCACTAA TCGTCTTCCG TATCGGATCA TTCATCCCGA TCCCGGGAAT 2161 CGACGCAGCA GTCCTAGCAA AACTACTAGA ACAGCAGCGT GGAACCATCA TCGAAATGTT 2221 CAACATGTTC TCAGGAGGAG CACTATCACG TGCATCAATC TTCGCACTAG GAATCATGCC 2281 GTACATCTCA GCATCAATCA TCATCCAGCT ACTAACCGTC GTCCACCCGA CCCTAGCAGA 2341 AATCAAAAAA GAAGGAGAAT CAGGACGTCG TAAAATCTCA CAGTACACCC GTTACGGAAC 2401 CCTAGTCCTA GCAATCTTCC AGTCAATCGG AATCGCAACC GGACTACCGA ACATGCCGGG 2461 AATGCAGGGA CTAGTCATCA ACCCGGGATT CGCATTCTAC TTCACCGCAG TCGTCTCACT 2521 AGTCACCGGA ACCATGTTCC TAATGTGGCT AGGAGAACAG ATCACCGAAC GTGGAATCGG 2581 AAACGGAATC TCAATCATCA TCTTCGCAGG AATCGTCGCA GGACTACCGC CGGCAATCGC 2641 ACACACCATC GAACAGGCAC GTCAGGGAGA CCTACACTTC CTAGTCCTAC TACTAGTCGC 2701 AGTCCTAGTC TTCGCAGTCA CCTTCTTCGT CGTCTTCGTC GAACGTGGAC AGCGTCGTAT 2761 CGTCGTCAAC TACGCAAAAC GTCAGCAGGG ACGTCGTGTC TACGCAGCAC AGTCAACCCA 2821 CCTACCGCTA AAAGTCAACA TGGCAGGAGT CATCCCGGCA ATCTTCGCAT CATCAATCAT 2881 CCTATTCCCG GCAACCATCG CATCATGGTT CGGAGGAGGA ACCGGATGGA ACTGGCTAAC 2941 CACCATCTCA CTATACCTAC AGCCGGGACA GCCGCTATAC GTCCTACTAT ACGCATCAGC 3001 AATCATCTTC TTCTGCTTCT TCTACACCGC ACTAGTCTTC AACCCGCGTG AAACCGCAGA 3061 CAACCTAAAA AAATCAGGAG CATTCGTCCC GGGAATCCGT CCGGGAGAAC AGACCGCAAA 3121 ATACATCGAC AAAGTCATGA CCCGTCTAAC CCTAGTCGGA GCACTATACA TCACCTTCAT 3181 CTGCCTAATC CCGGAATTCA TGCGTGACGC AATGAAAGTC CCGTTCTACT TCGGAGGAAC 3241 CTCACTACTA ATCGTCGTCG TCGTCATCAT GGACTTCATG GCACAGGTCC AGACCCTAAT 3301 GATGTCATCA CAGTACGAAT CAGCACTAAA AAAAGCAAAC CTAAAAGGAT ACGGACGTTG 3361 AAGAGCTTCG AGACCAATGC AAAGCTATTA GCGCGCGTCG AGTCGCCCGT AATACGTAGG 3421 GATAACAGGG TAATGCTCGG ATTTTTTGCA TCGGC //