LOCUS Exported 4628 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..4628 /label="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 2665..2712 /label="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind 4560..4576 /label="M13 for" /note="common sequencing primer, one of multiple similar variants" terminator 4577..4608 /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 2548..2567 /label="dialout_rev" /note="Forward dialout primer. Unique pair for each FreeGenes part." CDS 29..2533 /label="Tth DNA polymerase" /note="" /translation="MEAMLPLFEPKGRVLLVDGHHLAYRTFFALKGLTTSRGEPVQAVYGFAKSLLKALKEDGYKAVFVVFDAKAPSFRHEAYEAYKAGRAPTPEDFPRQLALIKELVDLLGFTRLEVPGYEADDVLATLAKKAEKEGYEVRILTADRDLYQLVSDRVAVLHPEGHLITPEWLWEKYGLRPEQWVDFRALVGDPSDNLPGVKGIGEKTALKLLKEWGSLENLLKNLDRVKPENVREKIKAHLEDLRLSLELSRVRTDLPLEVDLAQGREPDREGLRAFLERLEFGSLLHEFGLLEAPAPLEEAPWPPPEGAFVGFVLSRPEPMWAELKALAACRDGRVHRAADPLAGLKDLKEVRGLLAKDLAVLASREGLDLVPGDDPMLLAYLLDPSNTTPEGVARRYGGEWTEDAAHRALLSERLHRNLLKRLEGEEKLLWLYHEVEKPLSRVLAHMEATGVRRDVAYLQALSLELAEEIRRLEEEVFRLAGHPFNLNSRDQLERVLFDELRLPALGKTQKTGKRSTSAAVLEALREAHPIVEKILQHRELTKLKNTYVDPLPSLVHPRTGRLHTRFNQTATATGRLSSSDPNLQNIPVRTPLGQRIRRAFVAEAGWALVALDYSQIELRVLAHLSGDENLIRVFQEGKDIHTQTASWMFGVPPEAVDPLMRRAAKTVNFGVLYGMSAHRLSQELAIPYEEAVAFIERYFQSFPKVRAWIEKTLEEGRKRGYVETLFGRRRYVPDLNARVKSVREAAERMAFNMPVQGTAADLMKLAMVKLFPRLREMGARMLLQVHDELLLEAPQARAEEVAALAKEAMEKAYPLAVPLEVEVGMGEDWLSAKG*" primer_bind complement(2717..2733) /label="M13 rev" /note="common sequencing primer, one of multiple similar variants" rep_origin complement(2800..3388) /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(3579..4439) /label="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*" promoter complement(4440..4544) /label="bla promoter" /note="ampicillin resistance gene promoter" misc_feature 2568..2627 /label="FreeGenes Barcode" /note="FreeGenes Barcode. Defined as I-SceI site flanked by 21 base pair random sequences." ORIGIN 1 AAAGGGGCCG TCAATATCAG GGTCTCTAAT GGAGGCTATG CTGCCGTTGT TCGAACCGAA 61 AGGCCGCGTG CTGCTGGTTG ATGGCCATCA CCTGGCCTAC CGTACCTTCT TTGCGCTGAA 121 GGGGCTGACT ACTTCTCGTG GTGAACCGGT GCAAGCCGTG TACGGTTTCG CCAAATCCCT 181 GCTGAAAGCT CTGAAAGAGG ACGGCTACAA AGCGGTATTC GTGGTATTCG ATGCGAAAGC 241 GCCGTCTTTT CGTCATGAAG CGTACGAAGC TTACAAAGCA GGCCGTGCTC CGACCCCTGA 301 GGACTTCCCG CGTCAGCTGG CCCTGATTAA AGAACTGGTC GACCTGCTGG GTTTCACCCG 361 TCTGGAGGTT CCGGGCTACG AAGCCGATGA CGTCCTGGCA ACTCTCGCTA AAAAAGCTGA 421 AAAAGAAGGT TATGAAGTTC GTATCCTGAC GGCCGACCGC GACCTGTACC AGCTGGTTTC 481 TGACCGTGTG GCTGTGTTGC ACCCGGAAGG TCATCTGATT ACCCCGGAAT GGCTTTGGGA 541 AAAATACGGT CTGCGTCCGG AACAGTGGGT GGACTTCCGT GCGCTGGTTG GCGATCCGTC 601 CGACAACCTG CCGGGTGTTA AAGGCATTGG TGAAAAAACG GCTCTCAAAC TGCTGAAAGA 661 GTGGGGGTCC CTGGAGAACC TGCTGAAAAA CTTGGATCGC GTAAAACCGG AGAACGTCCG 721 TGAAAAAATT AAAGCGCACC TGGAGGACCT GCGTCTGTCC CTGGAGCTCT CTCGTGTGCG 781 CACTGACCTC CCGCTGGAAG TGGATCTCGC TCAGGGTCGT GAACCGGACC GTGAGGGCCT 841 GCGTGCGTTT CTGGAAAGAC TGGAGTTTGG TTCCCTCCTG CACGAGTTCG GCCTGCTCGA 901 AGCGCCGGCA CCGCTCGAAG AGGCTCCTTG GCCGCCGCCG GAAGGCGCTT TCGTAGGCTT 961 CGTTCTGAGC CGTCCGGAGC CGATGTGGGC AGAACTTAAG GCGCTGGCTG CTTGCCGTGA 1021 TGGTCGCGTC CACCGTGCCG CGGACCCGCT GGCAGGGCTG AAAGATTTAA AAGAAGTTCG 1081 CGGTCTGCTG GCTAAAGACC TCGCAGTGCT GGCGAGCCGT GAAGGACTGG ATCTGGTCCC 1141 GGGTGATGAT CCGATGCTGC TGGCGTACCT GCTGGACCCG TCCAACACCA CCCCGGAAGG 1201 CGTTGCGCGC CGTTACGGCG GAGAATGGAC CGAAGATGCA GCTCATCGTG CGCTGCTGTC 1261 TGAACGCCTG CATCGTAACC TGCTCAAACG CCTGGAAGGC GAAGAAAAAC TGCTGTGGCT 1321 TTATCACGAA GTGGAAAAAC CTCTGTCTCG TGTACTGGCC CACATGGAAG CGACCGGCGT 1381 GCGTCGTGAT GTGGCTTACC TGCAAGCCCT GAGCCTGGAG CTGGCTGAAG AAATCCGCCG 1441 TCTGGAAGAA GAGGTGTTCC GTCTGGCGGG TCATCCTTTT AACCTGAACA GCCGTGACCA 1501 GCTGGAACGC GTCCTGTTTG ACGAACTGCG TCTGCCGGCT CTGGGCAAGA CCCAGAAAAC 1561 CGGCAAACGT AGCACTTCCG CTGCTGTGCT TGAAGCACTG CGTGAGGCCC ATCCGATCGT 1621 CGAAAAAATC CTGCAACACC GTGAACTGAC GAAACTGAAA AACACCTACG TGGACCCGCT 1681 GCCGAGCCTG GTACATCCGC GTACCGGTCG GCTGCACACC CGTTTTAATC AGACCGCTAC 1741 CGCAACCGGT CGTCTTTCTA GCTCTGACCC GAACCTGCAA AACATCCCGG TACGTACCCC 1801 GCTGGGTCAG CGTATCCGTC GTGCTTTCGT CGCGGAAGCC GGTTGGGCCT TGGTCGCCCT 1861 GGATTATAGT CAGATCGAAC TGCGTGTGCT TGCGCACCTG TCTGGCGACG AAAACTTAAT 1921 TCGTGTGTTC CAGGAGGGTA AAGACATCCA CACCCAGACG GCATCGTGGA TGTTCGGCGT 1981 ACCGCCGGAA GCGGTTGACC CGCTGATGCG CCGCGCAGCC AAAACCGTTA ACTTCGGTGT 2041 CCTGTACGGT ATGTCTGCGC ACCGTCTGTC CCAGGAGCTG GCGATCCCCT ACGAAGAGGC 2101 AGTTGCGTTT ATCGAACGTT ACTTTCAGAG CTTCCCGAAA GTGCGTGCTT GGATCGAAAA 2161 AACCCTGGAA GAAGGTCGTA AACGCGGCTA CGTCGAAACC CTGTTTGGTC GCCGTCGTTA 2221 TGTGCCGGAC CTGAACGCTC GTGTGAAGAG TGTTCGTGAA GCCGCCGAGC GTATGGCGTT 2281 TAACATGCCG GTTCAGGGGA CCGCGGCTGA CCTGATGAAA CTGGCAATGG TGAAACTCTT 2341 CCCGCGTCTG CGTGAAATGG GTGCTCGCAT GCTTCTGCAA GTGCACGATG AATTGCTCCT 2401 GGAAGCTCCG CAGGCGCGTG CGGAGGAAGT GGCAGCCCTT GCGAAAGAAG CTATGGAAAA 2461 AGCCTACCCG CTGGCAGTTC CGCTGGAAGT CGAAGTGGGT ATGGGTGAAG ATTGGCTCTC 2521 CGCGAAAGGC TGAAGAGCTT TGAGACCACT AGATTAGCAA GGCACCCGTG TGCAAACACT 2581 TGCTTCTCTA GGGATAACAG GGTAATGGGA CAGAGGTCGT AGTCGCCCGC TGAGGTGTCA 2641 ATCGTCGGAG CCGCTGAGCA ATAACTAGCA TAACCCCTTG GGGCCTCTAA ACGGGTCTTG 2701 AGGGGTTTTT TGCATGGTCA TAGCTGTTTC CTGAGAGCTT GGCAGGTGAT GACACACATT 2761 AACAAATTTC GTGAGGAGTC TCCAGAAGAA TGCCATTAAT TTCCATAGGC TCCGCCCCCC 2821 TGACGAGCAT CACAAAAATC GACGCTCAAG TCAGAGGTGG CGAAACCCGA CAGGACTATA 2881 AAGATACCAG GCGTTTCCCC CTGGAAGCTC CCTCGTGCGC TCTCCTGTTC CGACCCTGCC 2941 GCTTACCGGA TACCTGTCCG CCTTTCTCCC TTCGGGAAGC GTGGCGCTTT CTCATAGCTC 3001 ACGCTGTAGG TATCTCAGTT CGGTGTAGGT CGTTCGCTCC AAGCTGGGCT GTGTGCACGA 3061 ACCCCCCGTT CAGCCCGACC GCTGCGCCTT ATCCGGTAAC TATCGTCTTG AGTCCAACCC 3121 GGTAAGACAC GACTTATCGC CACTGGCAGC AGCCACTGGT AACAGGATTA GCAGAGCGAG 3181 GTATGTAGGC GGTGCTACAG AGTTCTTGAA GTGGTGGCCT AACTACGGCT ACACTAGAAG 3241 AACAGTATTT GGTATCTGCG CTCTGCTGAA GCCAGTTACC TTCGGAAAAA GAGTTGGTAG 3301 CTCTTGATCC GGCAAACAAA CCACCGCTGG TAGCGGTGGT TTTTTTGTTT GCAAGCAGCA 3361 GATTACGCGC AGAAAAAAAG GATCTCAAGA AGGCCTACTA TTAGCAACAA CGATCCTTTG 3421 ATCTTTTCTA CGGGGTCTGA CGCTCAGTGG AACGAAAACT CACGTTAAGG GATTTTGGTC 3481 ATGAGATTAT CAAAAAGGAT CTTCACCTAG ATCCTTTTAA ATTAAAAATG AAGTTTTAAA 3541 TCAATCTAAA GTATATATGA GTAAACTTGG TCTGACAGTT ACCAATGCTT AATCAGTGAG 3601 GCACCTATCT CAGCGATCTG TCTATTTCGT TCATCCATAG TTGCCTGACT CCCCGTCGTG 3661 TAGATAACTA CGATACGGGA GGGCTTACCA TCTGGCCCCA GTGCTGCAAT GATACCGCGA 3721 GAACCACGCT CACCGGCTCC AGATTTATCA GCAATAAACC AGCCAGCCGG AAGGGCCGAG 3781 CGCAGAAGTG GTCCTGCAAC TTTATCCGCC TCCATCCAGT CTATTAATTG TTGCCGGGAA 3841 GCTAGAGTAA GTAGTTCGCC AGTTAATAGT TTGCGCAACG TTGTTGCCAT TGCTACAGGC 3901 ATCGTGGTGT CACGCTCGTC GTTTGGTATG GCTTCATTCA GCTCCGGTTC CCAACGATCA 3961 AGGCGAGTTA CATGATCCCC CATGTTGTGC AAAAAAGCGG TTAGCTCCTT CGGTCCTCCG 4021 ATCGTTGTCA GAAGTAAGTT GGCCGCAGTG TTATCACTCA TGGTTATGGC AGCACTGCAT 4081 AATTCTCTTA CTGTCATGCC ATCCGTAAGA TGCTTTTCTG TGACTGGTGA GTACTCAACC 4141 AAGTCATTCT GAGAATAGTG TATGCGGCGA CCGAGTTGCT CTTGCCCGGC GTCAATACGG 4201 GATAATACCG CGCCACATAG CAGAACTTTA AAAGTGCTCA TCATTGGAAA ACGTTCTTCG 4261 GGGCGAAAAC TCTCAAGGAT CTTACCGCTG TTGAGATCCA GTTCGATGTA ACCCACTCGT 4321 GCACCCAACT GATCTTCAGC ATCTTTTACT TTCACCAGCG TTTCTGGGTG AGCAAAAACA 4381 GGAAGGCAAA ATGCCGCAAA AAAGGGAATA AGGGCGACAC GGAAATGTTG AATACTCATA 4441 CTCTTCCTTT TTCAATATTA TTGAAGCATT TATCAGGGTT ATTGTCTCAT GAGCGGATAC 4501 ATATTTGAAT GTATTTAGAA AAATAAACAA ATAGGGGTTC CGCGCACCTG CACCAGTCAG 4561 TAAAACGACG GCCAGTAGTC AAAAGCCTCC GACCGGAGGC TTTTGACTTG GTTCAGGTGG 4621 AGTGGGAG //