LOCUS Exported 4622 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..4622 /label="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 2659..2706 /label="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind 4554..4570 /label="M13 for" /note="common sequencing primer, one of multiple similar variants" terminator 4571..4602 /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 2542..2561 /label="dialout_rev" /note="Forward dialout primer. Unique pair for each FreeGenes part." CDS 29..2527 /translation="MRGMLPLFEPKGRVLLVDGHHLAYRTFHALKGLTTSRGEPVQAVYGFAKSLLKALKEDGDAVIVVFDAKAPSFRHEAYGGYKAGRAPTPEDFPRQLALIKELVDLLGLARLEVPGYEADDVLASLAKKAEKEGYEVRILTADKDLYQLLSDRIHVLHPEGYLITPAWLWEKYGLRPDQWADYRALTGDESDNLPGVKGIGEKTARKLLEEWGSLEALLKNLDRLKPAIREKILAHMDDLKLSWDLAKVRTDLPLEVDFAKRREPDRERLRAFLERLEFGSLLHEFGLLESPKALEEAPWPPPEGAFVGFVLSRKEPMWADLLALAAARGGRVHRAPEPYKALRDLKEARGLLAKDLSVLALREGLGLPPGDDPMLLAYLLDPSNTTPEGVARRYGGEWTEEAGERAALSERLFANLWGRLEGEERLLWLYREVERPLSAVLAHMEATGVRLDVAYLRALSLEVAEEIARLEAEVFRLAGHPFNLNSRDQLERVLFDELGLPAIGKTEKTGKRSTSAAVLEALREAHPIVEKILQYRELTKLKSTYIDPLPDLIHPRTGRLHTRFNQTATATGRLSSSDPNLQNIPVRTPLGQRIRRAFIAEEGWLLVALDYSQIELRVLAHLSGDENLIRVFQEGRDIHTETASWMFGVPREAVDPLMRRAAKTINFGVLYGMSAHRLSQELAIPYEEAQAFIERYFQSFPKVRAWIEKTLEEGRRRGYVETLFGRRRYVPDLEARVKSVREAAERMAFNMPVQGTAADLMKLAMVKLFPRLEEMGARMLLQVHDELVLEAPKERAEAVARLAKEVMEGVYPLAVPLEVEVGIGEDWLSAKE*" /label="Taq DNA Polymerase" /note="" primer_bind complement(2711..2727) /label="M13 rev" /note="common sequencing primer, one of multiple similar variants" rep_origin complement(2794..3382) /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3573..4433) /label="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*" promoter complement(4434..4538) /label="bla promoter" /note="ampicillin resistance gene promoter" misc_feature 2562..2621 /label="FreeGenes Barcode" /note="FreeGenes Barcode. Defined as I-SceI site flanked by 21 base pair random sequences." ORIGIN 1 AAAGGGGCCG TCAATATCAG GGTCTCTAAT GCGTGGTATG CTGCCGCTGT TCGAACCGAA 61 AGGTCGTGTT CTTCTGGTTG ATGGTCATCA TCTGGCGTAC CGTACCTTCC ATGCGCTGAA 121 GGGACTGACG ACTTCTCGTG GCGAACCTGT ACAGGCCGTG TACGGTTTCG CCAAAAGCCT 181 GTTGAAAGCG CTGAAAGAAG ATGGTGATGC GGTTATTGTT GTTTTTGATG CGAAGGCTCC 241 GTCCTTCCGT CACGAAGCAT ACGGTGGTTA CAAAGCAGGC CGCGCGCCTA CTCCGGAAGA 301 TTTCCCGCGT CAGCTGGCCC TGATCAAAGA ACTGGTGGAT CTGCTTGGTC TGGCGCGTCT 361 GGAAGTCCCG GGTTATGAAG CTGATGATGT ACTGGCGTCC CTGGCGAAAA AAGCTGAAAA 421 AGAAGGTTAC GAAGTGCGTA TCCTGACCGC CGATAAAGAC CTGTACCAAC TGCTGTCCGA 481 CCGTATCCAC GTACTGCACC CGGAAGGCTA CCTGATCACC CCAGCTTGGC TGTGGGAGAA 541 GTACGGCCTG CGTCCGGATC AGTGGGCCGA CTATCGCGCA CTGACCGGTG ATGAATCAGA 601 TAACCTGCCG GGTGTCAAGG GCATTGGTGA AAAAACCGCA CGCAAGCTGT TGGAAGAGTG 661 GGGGAGCCTG GAAGCACTGT TGAAAAATCT GGATCGCCTG AAACCAGCAA TCCGTGAAAA 721 GATCCTGGCA CATATGGATG ATCTGAAACT GTCGTGGGAC CTGGCTAAAG TGCGTACCGA 781 CCTGCCGCTG GAGGTGGACT TTGCGAAACG CCGCGAACCG GATCGTGAAC GTCTGCGCGC 841 TTTCCTGGAA CGTCTGGAGT TCGGTTCTCT CTTACACGAG TTCGGCCTGC TGGAATCGCC 901 GAAGGCCCTG GAAGAAGCGC CGTGGCCGCC GCCCGAGGGT GCGTTTGTAG GCTTCGTGCT 961 GTCTCGCAAA GAACCGATGT GGGCAGACCT GCTGGCACTG GCGGCGGCCC GCGGCGGTCG 1021 TGTTCACCGT GCACCGGAAC CATACAAAGC GCTGCGTGAC CTGAAAGAAG CGCGCGGCCT 1081 GCTGGCAAAG GATCTGAGCG TGCTCGCCCT TCGTGAAGGC CTGGGTCTGC CGCCGGGCGA 1141 CGATCCGATG CTCTTAGCGT ACTTACTGGA CCCGTCTAAC ACCACCCCGG AAGGTGTCGC 1201 GCGTCGTTAC GGCGGCGAAT GGACTGAAGA AGCGGGTGAA CGTGCAGCTC TGAGCGAACG 1261 TCTGTTCGCA AATCTGTGGG GTCGTTTGGA GGGTGAGGAA CGCCTGCTGT GGCTGTACCG 1321 TGAAGTAGAG CGTCCGCTGT CTGCGGTTCT GGCTCACATG GAGGCGACTG GCGTTCGCCT 1381 GGATGTTGCT TATCTTCGTG CGCTGAGTCT GGAAGTGGCT GAAGAGATCG CACGTCTGGA 1441 GGCAGAAGTC TTTCGTCTTG CAGGTCATCC GTTTAACCTG AACTCCCGCG ATCAGCTGGA 1501 ACGCGTACTG TTTGACGAGC TGGGTCTGCC GGCTATCGGT AAAACTGAAA AAACCGGTAA 1561 GCGTTCCACG AGTGCAGCAG TCTTGGAGGC GCTCCGCGAA GCTCACCCGA TTGTGGAAAA 1621 AATTCTGCAA TACCGTGAAC TCACTAAACT GAAGTCCACC TATATTGATC CGCTGCCGGA 1681 CCTGATCCAC CCTCGTACTG GTCGTTTACA TACCCGTTTT AACCAGACCG CTACGGCAAC 1741 CGGTCGTCTG AGTTCCTCTG ACCCGAACCT CCAGAATATC CCGGTTCGTA CCCCGCTGGG 1801 TCAGCGTATC CGCCGCGCAT TCATTGCCGA GGAAGGCTGG CTGCTGGTAG CACTCGACTA 1861 TTCCCAGATT GAACTTCGTG TTCTGGCTCA TTTGTCTGGC GACGAAAACC TGATCCGCGT 1921 GTTTCAGGAA GGTCGTGACA TCCATACCGA GACTGCTTCG TGGATGTTCG GAGTCCCGCG 1981 TGAAGCCGTT GATCCGCTGA TGCGCCGCGC GGCCAAGACT ATCAACTTCG GTGTTCTGTA 2041 CGGTATGTCT GCGCACCGTC TGTCCCAGGA GCTGGCCATT CCGTACGAAG AAGCACAAGC 2101 GTTCATCGAG CGTTATTTTC AATCCTTCCC GAAAGTCCGC GCATGGATTG AAAAAACCCT 2161 GGAAGAGGGC CGCCGCCGTG GTTACGTGGA AACCCTGTTT GGCCGCCGTC GCTACGTGCC 2221 GGACCTGGAG GCCCGCGTTA AATCCGTGCG CGAGGCAGCG GAACGTATGG CGTTCAACAT 2281 GCCTGTTCAG GGTACGGCAG CCGACCTGAT GAAACTGGCC ATGGTTAAAC TGTTCCCGCG 2341 TCTGGAAGAA ATGGGCGCGC GTATGCTGCT GCAAGTACAC GACGAACTGG TCTTGGAAGC 2401 GCCAAAGGAA CGTGCGGAAG CGGTTGCTCG TCTGGCAAAA GAGGTCATGG AGGGGGTGTA 2461 TCCGCTGGCG GTACCGCTGG AAGTTGAAGT TGGCATCGGC GAAGATTGGC TGTCTGCTAA 2521 AGAGTGAAGA GCTTTGAGAC CACGTATGGG GAACACTACA CAACCCCTGG TTACAACTCT 2581 CGTAGGGATA ACAGGGTAAT GACACGAGCC CCATGCCCCA ACGCTGAGGT GTCAATCGTC 2641 GGAGCCGCTG AGCAATAACT AGCATAACCC CTTGGGGCCT CTAAACGGGT CTTGAGGGGT 2701 TTTTTGCATG GTCATAGCTG TTTCCTGAGA GCTTGGCAGG TGATGACACA CATTAACAAA 2761 TTTCGTGAGG AGTCTCCAGA AGAATGCCAT TAATTTCCAT AGGCTCCGCC CCCCTGACGA 2821 GCATCACAAA AATCGACGCT CAAGTCAGAG GTGGCGAAAC CCGACAGGAC TATAAAGATA 2881 CCAGGCGTTT CCCCCTGGAA GCTCCCTCGT GCGCTCTCCT GTTCCGACCC TGCCGCTTAC 2941 CGGATACCTG TCCGCCTTTC TCCCTTCGGG AAGCGTGGCG CTTTCTCATA GCTCACGCTG 3001 TAGGTATCTC AGTTCGGTGT AGGTCGTTCG CTCCAAGCTG GGCTGTGTGC ACGAACCCCC 3061 CGTTCAGCCC GACCGCTGCG CCTTATCCGG TAACTATCGT CTTGAGTCCA ACCCGGTAAG 3121 ACACGACTTA TCGCCACTGG CAGCAGCCAC TGGTAACAGG ATTAGCAGAG CGAGGTATGT 3181 AGGCGGTGCT ACAGAGTTCT TGAAGTGGTG GCCTAACTAC GGCTACACTA GAAGAACAGT 3241 ATTTGGTATC TGCGCTCTGC TGAAGCCAGT TACCTTCGGA AAAAGAGTTG GTAGCTCTTG 3301 ATCCGGCAAA CAAACCACCG CTGGTAGCGG TGGTTTTTTT GTTTGCAAGC AGCAGATTAC 3361 GCGCAGAAAA AAAGGATCTC AAGAAGGCCT ACTATTAGCA ACAACGATCC TTTGATCTTT 3421 TCTACGGGGT CTGACGCTCA GTGGAACGAA AACTCACGTT AAGGGATTTT GGTCATGAGA 3481 TTATCAAAAA GGATCTTCAC CTAGATCCTT TTAAATTAAA AATGAAGTTT TAAATCAATC 3541 TAAAGTATAT ATGAGTAAAC TTGGTCTGAC AGTTACCAAT GCTTAATCAG TGAGGCACCT 3601 ATCTCAGCGA TCTGTCTATT TCGTTCATCC ATAGTTGCCT GACTCCCCGT CGTGTAGATA 3661 ACTACGATAC GGGAGGGCTT ACCATCTGGC CCCAGTGCTG CAATGATACC GCGAGAACCA 3721 CGCTCACCGG CTCCAGATTT ATCAGCAATA AACCAGCCAG CCGGAAGGGC CGAGCGCAGA 3781 AGTGGTCCTG CAACTTTATC CGCCTCCATC CAGTCTATTA ATTGTTGCCG GGAAGCTAGA 3841 GTAAGTAGTT CGCCAGTTAA TAGTTTGCGC AACGTTGTTG CCATTGCTAC AGGCATCGTG 3901 GTGTCACGCT CGTCGTTTGG TATGGCTTCA TTCAGCTCCG GTTCCCAACG ATCAAGGCGA 3961 GTTACATGAT CCCCCATGTT GTGCAAAAAA GCGGTTAGCT CCTTCGGTCC TCCGATCGTT 4021 GTCAGAAGTA AGTTGGCCGC AGTGTTATCA CTCATGGTTA TGGCAGCACT GCATAATTCT 4081 CTTACTGTCA TGCCATCCGT AAGATGCTTT TCTGTGACTG GTGAGTACTC AACCAAGTCA 4141 TTCTGAGAAT AGTGTATGCG GCGACCGAGT TGCTCTTGCC CGGCGTCAAT ACGGGATAAT 4201 ACCGCGCCAC ATAGCAGAAC TTTAAAAGTG CTCATCATTG GAAAACGTTC TTCGGGGCGA 4261 AAACTCTCAA GGATCTTACC GCTGTTGAGA TCCAGTTCGA TGTAACCCAC TCGTGCACCC 4321 AACTGATCTT CAGCATCTTT TACTTTCACC AGCGTTTCTG GGTGAGCAAA AACAGGAAGG 4381 CAAAATGCCG CAAAAAAGGG AATAAGGGCG ACACGGAAAT GTTGAATACT CATACTCTTC 4441 CTTTTTCAAT ATTATTGAAG CATTTATCAG GGTTATTGTC TCATGAGCGG ATACATATTT 4501 GAATGTATTT AGAAAAATAA ACAAATAGGG GTTCCGCGCA CCTGCACCAG TCAGTAAAAC 4561 GACGGCCAGT AGTCAAAAGC CTCCGACCGG AGGCTTTTGA CTTGGTTCAG GTGGAGTGGG 4621 AG //