LOCUS Exported 4181 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..4181 /label="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 2218..2265 /label="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind 4113..4129 /label="M13 for" /note="common sequencing primer, one of multiple similar variants" terminator 4130..4161 /label="tonB terminator" /note="bidirectional E. coli tonB-P14 transcription terminator" primer_bind 1..20 /label="dialout_for" /note="Reverse dialout primer. Unique pair for each FreeGenes part." primer_bind 2101..2120 /label="dialout_rev" /note="Forward dialout primer. Unique pair for each FreeGenes part." CDS 29..2086 /translation="MGVGEKTAIKLLKQFDSVEKLLESIDEVSGKKLKEKLEEFKDQALMSKELATIMTDAPIEVSVSGLEYQGFNREQVIAIFKDLGFNTLLERLGEDSAEAEQDQSLEDINVKTVTDVTSDILVSPSAFVVEQIGDNYHEEPILGFSIVNETGAYFIPKDIAVESEVFKEWVENDEQKKWVFDSKRAVVALRWQGIELKGAEFDTLLAAYIINPGNSYDDVASVAKDYGLHIVSSDESVYGKGAKRAVPSEDVLSEHLGRKALAIQSLREKLVQELENNDQLELFEELEMPLALILGEMESTGVKVDVDRLKRMGEELGAKLKEYEEKIHEIAGEPFNINSPKQLGVILFEKIGLPVVKKTKTGYSTSADVLEKLADKHDIVDYILQYRQIGKLQSTYIEGLLKVTRPDSHKVHTRFNQALTQTGRLSSTDPNLQNIPIRLEEGRKIRQAFVPSEKDWLIFAADYSQIELRVLAHISKDENLIEAFTNDMDIHTKTAMDVFHVAKDEVTSAMRRQAKAVNFGIVYGISDYGLSQNLGITRKEAGAFIDRYLESFQGVKAYMEDSVQEAKQKGYVTTLMHRRRYIPELTSRNFNIRSFAERTAMNTPIQGSAADIIKKAMIDMAAKLKEKQLKARLLLQVHDELIFEAPKEEIEILEKLVPEVMEHALALDVPLKVDFASGPSWYDAK*" /label="Bsu DNA Polymerase I, Large Fragment " /note="" primer_bind complement(2270..2286) /label="M13 rev" /note="common sequencing primer, one of multiple similar variants" rep_origin complement(2353..2941) /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3132..3992) /label="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*" promoter complement(3993..4097) /note="ampicillin resistance gene promoter" /label="bla promoter" misc_feature 2121..2180 /label="FreeGenes Barcode" /note="FreeGenes Barcode. Defined as I-SceI site flanked by 21 base pair random sequences." ORIGIN 1 AAAGGGGCCG TCAATATCAG GGTCTCTAAT GGGCGTTGGC GAGAAAACCG CCATCAAACT 61 GCTGAAACAG TTCGATTCTG TCGAAAAATT ACTGGAAAGC ATTGACGAAG TCAGCGGTAA 121 GAAACTGAAA GAAAAGCTGG AGGAGTTCAA AGACCAAGCG CTGATGTCGA AAGAGCTGGC 181 AACCATCATG ACCGACGCCC CGATTGAAGT ATCTGTCTCC GGTCTGGAAT ACCAGGGCTT 241 CAACCGTGAG CAGGTTATTG CCATTTTTAA AGACCTGGGC TTCAACACCC TGCTGGAGCG 301 TCTGGGGGAA GATTCCGCTG AAGCGGAACA GGACCAGTCC CTGGAGGACA TTAACGTGAA 361 AACCGTGACT GATGTCACCA GCGATATTCT GGTCAGCCCG TCTGCCTTTG TTGTTGAGCA 421 AATCGGCGAC AACTACCACG AAGAACCGAT CCTGGGGTTC TCAATCGTTA ACGAAACCGG 481 TGCTTACTTC ATACCGAAAG ATATTGCGGT TGAATCTGAG GTGTTTAAAG AATGGGTAGA 541 GAACGATGAG CAGAAGAAAT GGGTTTTCGA CAGCAAGCGT GCTGTGGTCG CGTTGCGCTG 601 GCAGGGCATC GAACTGAAAG GTGCTGAGTT TGACACCCTG CTGGCGGCTT ATATCATCAA 661 CCCGGGTAAC TCTTACGATG ACGTTGCCTC TGTGGCAAAA GACTACGGCC TGCATATTGT 721 GAGCTCTGAT GAAAGCGTTT ATGGCAAGGG CGCAAAACGC GCGGTGCCGT CCGAGGACGT 781 GCTGTCCGAA CACCTGGGTC GCAAAGCTCT GGCTATTCAG TCCCTGCGTG AAAAACTGGT 841 CCAGGAGTTA GAAAACAACG ACCAACTCGA ACTGTTCGAA GAACTGGAAA TGCCGCTGGC 901 CCTGATCCTG GGTGAGATGG AATCTACCGG TGTAAAAGTG GACGTTGACC GTCTGAAGCG 961 TATGGGCGAA GAGTTGGGTG CTAAACTGAA AGAATATGAG GAAAAAATCC ATGAAATCGC 1021 GGGCGAACCG TTCAATATTA ACTCCCCTAA ACAACTGGGT GTCATTCTGT TCGAAAAAAT 1081 CGGCCTGCCG GTCGTAAAAA AAACCAAAAC GGGTTATTCC ACTAGCGCAG ATGTTTTAGA 1141 AAAGCTTGCA GATAAACACG ATATTGTGGA TTACATCCTG CAATATCGTC AGATTGGCAA 1201 GCTTCAGTCT ACCTACATCG AAGGCCTGTT AAAGGTTACT CGTCCAGATT CTCACAAGGT 1261 GCACACCCGT TTCAACCAGG CACTGACCCA GACCGGCCGC CTGAGCTCTA CCGATCCGAA 1321 CCTGCAAAAC ATCCCAATCC GTCTGGAAGA GGGTCGTAAA ATCCGCCAGG CTTTCGTGCC 1381 GTCTGAAAAA GACTGGCTGA TTTTTGCGGC GGACTATAGT CAGATCGAGC TGCGCGTTCT 1441 GGCACATATC TCTAAGGACG AAAACTTGAT CGAGGCTTTC ACCAACGACA TGGATATTCA 1501 TACGAAAACC GCCATGGACG TATTTCACGT AGCCAAAGAC GAGGTAACCT CCGCTATGCG 1561 TCGCCAGGCG AAGGCTGTAA ACTTTGGCAT CGTTTACGGT ATCTCCGACT ACGGGTTGTC 1621 TCAGAACCTG GGCATCACTC GTAAGGAAGC CGGCGCTTTT ATTGATCGTT ACCTGGAATC 1681 CTTCCAGGGT GTGAAAGCAT ACATGGAAGA TTCCGTTCAG GAGGCGAAAC AAAAAGGCTA 1741 CGTCACTACC CTGATGCACC GCCGTCGTTA TATCCCGGAA CTGACCTCCC GTAACTTCAA 1801 CATTCGAAGC TTCGCGGAAC GTACGGCCAT GAACACTCCG ATCCAGGGTT CCGCAGCCGA 1861 CATCATCAAA AAGGCCATGA TCGATATGGC GGCGAAACTG AAAGAAAAGC AACTGAAGGC 1921 GCGTCTGTTG CTGCAAGTGC ATGACGAACT GATCTTCGAA GCGCCGAAAG AGGAAATTGA 1981 AATCCTGGAA AAACTGGTTC CGGAAGTGAT GGAACATGCG CTGGCGCTGG ATGTTCCGCT 2041 GAAAGTTGAT TTCGCTTCGG GCCCGTCCTG GTATGATGCG AAGTGAAGAG CTTTGAGACC 2101 AGAACATAGC ATTCACGGGG GTACAGTAGG GGAGGAATGA ATAGGGATAA CAGGGTAATT 2161 CCTCTTTCTA GTGAAGGGCC CGCTGAGGTG TCAATCGTCG GAGCCGCTGA GCAATAACTA 2221 GCATAACCCC TTGGGGCCTC TAAACGGGTC TTGAGGGGTT TTTTGCATGG TCATAGCTGT 2281 TTCCTGAGAG CTTGGCAGGT GATGACACAC ATTAACAAAT TTCGTGAGGA GTCTCCAGAA 2341 GAATGCCATT AATTTCCATA GGCTCCGCCC CCCTGACGAG CATCACAAAA ATCGACGCTC 2401 AAGTCAGAGG TGGCGAAACC CGACAGGACT ATAAAGATAC CAGGCGTTTC CCCCTGGAAG 2461 CTCCCTCGTG CGCTCTCCTG TTCCGACCCT GCCGCTTACC GGATACCTGT CCGCCTTTCT 2521 CCCTTCGGGA AGCGTGGCGC TTTCTCATAG CTCACGCTGT AGGTATCTCA GTTCGGTGTA 2581 GGTCGTTCGC TCCAAGCTGG GCTGTGTGCA CGAACCCCCC GTTCAGCCCG ACCGCTGCGC 2641 CTTATCCGGT AACTATCGTC TTGAGTCCAA CCCGGTAAGA CACGACTTAT CGCCACTGGC 2701 AGCAGCCACT GGTAACAGGA TTAGCAGAGC GAGGTATGTA GGCGGTGCTA CAGAGTTCTT 2761 GAAGTGGTGG CCTAACTACG GCTACACTAG AAGAACAGTA TTTGGTATCT GCGCTCTGCT 2821 GAAGCCAGTT ACCTTCGGAA AAAGAGTTGG TAGCTCTTGA TCCGGCAAAC AAACCACCGC 2881 TGGTAGCGGT GGTTTTTTTG TTTGCAAGCA GCAGATTACG CGCAGAAAAA AAGGATCTCA 2941 AGAAGGCCTA CTATTAGCAA CAACGATCCT TTGATCTTTT CTACGGGGTC TGACGCTCAG 3001 TGGAACGAAA ACTCACGTTA AGGGATTTTG GTCATGAGAT TATCAAAAAG GATCTTCACC 3061 TAGATCCTTT TAAATTAAAA ATGAAGTTTT AAATCAATCT AAAGTATATA TGAGTAAACT 3121 TGGTCTGACA GTTACCAATG CTTAATCAGT GAGGCACCTA TCTCAGCGAT CTGTCTATTT 3181 CGTTCATCCA TAGTTGCCTG ACTCCCCGTC GTGTAGATAA CTACGATACG GGAGGGCTTA 3241 CCATCTGGCC CCAGTGCTGC AATGATACCG CGAGAACCAC GCTCACCGGC TCCAGATTTA 3301 TCAGCAATAA ACCAGCCAGC CGGAAGGGCC GAGCGCAGAA GTGGTCCTGC AACTTTATCC 3361 GCCTCCATCC AGTCTATTAA TTGTTGCCGG GAAGCTAGAG TAAGTAGTTC GCCAGTTAAT 3421 AGTTTGCGCA ACGTTGTTGC CATTGCTACA GGCATCGTGG TGTCACGCTC GTCGTTTGGT 3481 ATGGCTTCAT TCAGCTCCGG TTCCCAACGA TCAAGGCGAG TTACATGATC CCCCATGTTG 3541 TGCAAAAAAG CGGTTAGCTC CTTCGGTCCT CCGATCGTTG TCAGAAGTAA GTTGGCCGCA 3601 GTGTTATCAC TCATGGTTAT GGCAGCACTG CATAATTCTC TTACTGTCAT GCCATCCGTA 3661 AGATGCTTTT CTGTGACTGG TGAGTACTCA ACCAAGTCAT TCTGAGAATA GTGTATGCGG 3721 CGACCGAGTT GCTCTTGCCC GGCGTCAATA CGGGATAATA CCGCGCCACA TAGCAGAACT 3781 TTAAAAGTGC TCATCATTGG AAAACGTTCT TCGGGGCGAA AACTCTCAAG GATCTTACCG 3841 CTGTTGAGAT CCAGTTCGAT GTAACCCACT CGTGCACCCA ACTGATCTTC AGCATCTTTT 3901 ACTTTCACCA GCGTTTCTGG GTGAGCAAAA ACAGGAAGGC AAAATGCCGC AAAAAAGGGA 3961 ATAAGGGCGA CACGGAAATG TTGAATACTC ATACTCTTCC TTTTTCAATA TTATTGAAGC 4021 ATTTATCAGG GTTATTGTCT CATGAGCGGA TACATATTTG AATGTATTTA GAAAAATAAA 4081 CAAATAGGGG TTCCGCGCAC CTGCACCAGT CAGTAAAACG ACGGCCAGTA GTCAAAAGCC 4141 TCCGACCGGA GGCTTTTGAC TTGGTTCAGG TGGAGTGGGA G //