LOCUS       Exported     4775 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..4775
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
 polymerase"
     terminator      2812..2859
                     /note="transcription terminator for bacteriophage T7 RNA polymerase"
                     /label="T7 terminator"
     primer_bind     4707..4723
                     /label="M13 for"
                     /note="common sequencing primer, one of multiple similar variants"
     terminator      4724..4755
                     /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     2695..2714
                     /label="dialout_rev"
                     /note="Forward dialout primer. Unique pair for each FreeGenes part."
     CDS             29..2680
                     /label="T7 RNA Polymerase"
                     /note=""
                     /translation="MNTINIAKNDFSDIELAAIPFNTLADHYGERLAREQLALEHESYEMGEARFRKMFERQLKAGEVADNAAAKPLITTLLPKMIARINDWFEEVKAKRGKRPTAFQFLQEIKPEAVAYITIKTTLACLTSADNTTVQAVASAIGRAIEDEARFGRIRDLEAKHFKKNVEEQLNKRVGHVYKKAFMQVVEADMLSKGLLGGEAWSSWHKEDSIHVGVRCIEMLIESTGMVSLHRQNAGVVGQDSETIELAPEYAEAIATRAGALAGISPMFQPCVVPPKPWTGITGGGYWANGRRPLALVRTHSKKALMRYEDVYMPEVYKAINIAQNTAWKINKKVLAVANVITKWKHCPVEDIPAIEREELPMKPEDIDMNPEALTAWKRAAAAVYRKDKARKSRRISLEFMLEQANKFANHKAIWFPYNMDWRGRVYAVSMFNPQGNDMTKGLLTLAKGKPIGKEGYYWLKIHGANCAGVDKVPFPERIKFIEENHENIMACAKSPLENTWWAEQDSPFCFLAFCFEYAGVQHHGLSYNCSLPLAFDGSCSGIQHFSAMLRDEVGGRAVNLLPSETVQDIYGIVAKKVNEILQADAINGTDNEVVTVTDENTGEISEKVKLGTKALAGQWLAYGVTRSVTKRSVMTLAYGSKEFGFRQQVLEDTIQPAIDSGKGLMFTQPNQAAGYMAKLIWESVSVTVVAAVEAMNWLKSAAKLLAAEVKDKKTGEILRKRCAVHWVTPDGFPVWQEYKKPIQTRLNLMFLGQFRLQPTINTNKDSEIDAHKQESGIAPNFVHSQDGSHLRKTVVWAHEKYGIESFALIHDSFGTIPADAANLFKAVRETMVDTYESCDVLADFYDQFADQLHESQLDKMPALPAKGNLNLRDILESDFAFA*"
     primer_bind     complement(2864..2880)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar variants"
     rep_origin      complement(2947..3535)
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
                     /label="ori"
     CDS             complement(3726..4586)
                     /label="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"
     promoter        complement(4587..4691)
                     /label="bla promoter"
                     /note="ampicillin resistance gene promoter"
     misc_feature    2715..2774
                     /label="FreeGenes Barcode"
                     /note="FreeGenes Barcode. Defined as I-SceI site flanked by 21 base pair random sequences."
ORIGIN
        1 AAAGGGGCCG TCAATATCAG GGTCTCTAAT GAACACTATC AACATTGCGA AGAACGATTT
       61 CTCTGACATC GAACTCGCGG CAATCCCGTT CAACACCCTG GCAGATCACT ACGGTGAACG
      121 TCTGGCCCGT GAACAGCTGG CTCTGGAACA CGAAAGCTAT GAAATGGGTG AGGCGCGTTT
      181 CCGTAAGATG TTCGAACGTC AGTTAAAAGC GGGTGAAGTT GCAGACAACG CCGCGGCGAA
      241 GCCGCTGATC ACCACCCTGC TGCCGAAAAT GATCGCTCGC ATCAACGATT GGTTCGAAGA
      301 AGTGAAAGCG AAACGTGGCA AACGTCCGAC TGCCTTCCAG TTCCTGCAAG AAATTAAACC
      361 GGAGGCAGTG GCGTATATCA CTATCAAAAC TACCCTGGCG TGCCTGACCT CTGCGGATAA
      421 CACGACCGTT CAGGCGGTAG CGTCCGCAAT CGGTCGTGCG ATCGAAGATG AAGCCCGCTT
      481 CGGCCGTATC CGTGACCTTG AAGCAAAACA CTTTAAAAAA AACGTTGAAG AACAGCTGAA
      541 CAAACGCGTT GGTCACGTAT ACAAAAAGGC GTTTATGCAG GTTGTGGAAG CTGATATGCT
      601 GTCCAAAGGC CTGCTGGGCG GCGAAGCGTG GAGCTCTTGG CATAAAGAAG ATTCTATTCA
      661 CGTGGGCGTT CGCTGCATTG AAATGCTGAT CGAATCGACT GGTATGGTTT CTCTGCACCG
      721 TCAGAACGCT GGTGTTGTTG GCCAGGATTC TGAAACTATC GAACTGGCCC CGGAATATGC
      781 GGAAGCGATC GCTACTCGCG CAGGTGCGCT GGCCGGTATT TCCCCGATGT TCCAGCCGTG
      841 CGTTGTACCG CCTAAACCGT GGACCGGCAT CACCGGCGGT GGTTATTGGG CTAACGGTCG
      901 CCGTCCGCTG GCACTCGTTC GTACCCACAG CAAAAAGGCA CTGATGCGTT ACGAAGATGT
      961 GTACATGCCG GAGGTTTACA AAGCCATCAA CATTGCTCAG AATACCGCCT GGAAGATTAA
     1021 TAAAAAAGTG CTCGCGGTGG CGAACGTTAT TACCAAGTGG AAACACTGTC CGGTGGAAGA
     1081 TATCCCGGCT ATCGAGCGCG AGGAACTGCC GATGAAACCG GAAGATATCG ATATGAACCC
     1141 GGAAGCTCTG ACCGCGTGGA AACGTGCTGC TGCTGCCGTA TATCGCAAAG ACAAGGCGCG
     1201 TAAGTCTCGC CGTATCTCTC TGGAGTTTAT GCTGGAACAG GCGAACAAAT TCGCCAACCA
     1261 CAAAGCCATC TGGTTCCCGT ACAACATGGA TTGGCGTGGC CGTGTCTATG CAGTTTCTAT
     1321 GTTCAACCCG CAAGGGAATG ATATGACCAA AGGCCTGCTG ACTCTGGCAA AAGGTAAGCC
     1381 GATCGGCAAA GAAGGTTATT ACTGGCTGAA AATCCATGGC GCGAATTGCG CGGGTGTAGA
     1441 TAAGGTACCG TTCCCGGAAC GTATTAAGTT CATTGAAGAG AACCACGAGA ATATCATGGC
     1501 ATGCGCGAAA AGCCCGCTGG AAAACACTTG GTGGGCTGAG CAGGACTCCC CGTTCTGCTT
     1561 CCTGGCGTTC TGCTTCGAGT ACGCAGGTGT TCAACATCAT GGCCTGAGCT ACAACTGCTC
     1621 CCTGCCTCTG GCATTCGACG GTTCTTGCAG CGGTATCCAG CACTTTTCCG CTATGTTACG
     1681 TGATGAAGTA GGTGGCCGTG CTGTCAATCT GCTGCCGTCC GAAACCGTTC AAGATATCTA
     1741 TGGTATCGTA GCGAAAAAGG TTAACGAGAT TCTGCAAGCA GATGCTATCA ACGGTACCGA
     1801 CAACGAGGTA GTTACCGTGA CCGATGAAAA CACCGGTGAA ATTTCCGAAA AAGTAAAACT
     1861 GGGGACCAAG GCACTGGCTG GCCAGTGGCT GGCCTACGGC GTGACTCGCA GCGTCACCAA
     1921 ACGTAGCGTG ATGACGCTCG CTTATGGCAG CAAAGAGTTT GGCTTTCGCC AGCAAGTTCT
     1981 GGAGGATACT ATCCAGCCAG CCATTGACTC GGGCAAGGGT CTGATGTTTA CCCAGCCGAA
     2041 CCAGGCCGCT GGATATATGG CAAAACTGAT CTGGGAATCT GTAAGCGTCA CCGTTGTGGC
     2101 AGCGGTTGAA GCTATGAATT GGTTGAAAAG CGCCGCGAAA CTGCTGGCAG CGGAAGTGAA
     2161 AGATAAAAAA ACTGGCGAAA TCCTGCGTAA ACGCTGCGCG GTTCACTGGG TCACTCCGGA
     2221 TGGCTTCCCA GTTTGGCAGG AATACAAAAA ACCTATCCAG ACCCGCCTGA ATCTGATGTT
     2281 CCTGGGCCAA TTCCGTCTGC AACCGACCAT CAACACCAAC AAAGACAGCG AAATTGACGC
     2341 CCACAAACAG GAGTCCGGCA TTGCCCCTAA CTTCGTTCAC TCTCAGGACG GCTCTCATCT
     2401 GCGCAAAACT GTCGTTTGGG CTCATGAAAA GTACGGCATC GAATCCTTTG CGCTGATCCA
     2461 CGACTCCTTC GGTACCATCC CGGCGGACGC GGCCAACCTG TTCAAAGCTG TTCGTGAAAC
     2521 TATGGTAGAC ACGTACGAAA GCTGCGACGT TCTGGCAGAC TTCTACGATC AGTTCGCTGA
     2581 CCAGCTGCAC GAAAGCCAGC TGGACAAGAT GCCAGCTCTG CCGGCTAAAG GTAATCTGAA
     2641 CCTGCGCGAC ATTCTGGAAT CTGACTTCGC TTTCGCCTGA AGAGCTTTGA GACCAGTCCG
     2701 ACACAATGTG ACACTCAGGT AAGTTTTATG ATTATTAGGG ATAACAGGGT AATGAAGCTA
     2761 GCGGATACGC GGAACGCTGA GGTGTCAATC GTCGGAGCCG CTGAGCAATA ACTAGCATAA
     2821 CCCCTTGGGG CCTCTAAACG GGTCTTGAGG GGTTTTTTGC ATGGTCATAG CTGTTTCCTG
     2881 AGAGCTTGGC AGGTGATGAC ACACATTAAC AAATTTCGTG AGGAGTCTCC AGAAGAATGC
     2941 CATTAATTTC CATAGGCTCC GCCCCCCTGA CGAGCATCAC AAAAATCGAC GCTCAAGTCA
     3001 GAGGTGGCGA AACCCGACAG GACTATAAAG ATACCAGGCG TTTCCCCCTG GAAGCTCCCT
     3061 CGTGCGCTCT CCTGTTCCGA CCCTGCCGCT TACCGGATAC CTGTCCGCCT TTCTCCCTTC
     3121 GGGAAGCGTG GCGCTTTCTC ATAGCTCACG CTGTAGGTAT CTCAGTTCGG TGTAGGTCGT
     3181 TCGCTCCAAG CTGGGCTGTG TGCACGAACC CCCCGTTCAG CCCGACCGCT GCGCCTTATC
     3241 CGGTAACTAT CGTCTTGAGT CCAACCCGGT AAGACACGAC TTATCGCCAC TGGCAGCAGC
     3301 CACTGGTAAC AGGATTAGCA GAGCGAGGTA TGTAGGCGGT GCTACAGAGT TCTTGAAGTG
     3361 GTGGCCTAAC TACGGCTACA CTAGAAGAAC AGTATTTGGT ATCTGCGCTC TGCTGAAGCC
     3421 AGTTACCTTC GGAAAAAGAG TTGGTAGCTC TTGATCCGGC AAACAAACCA CCGCTGGTAG
     3481 CGGTGGTTTT TTTGTTTGCA AGCAGCAGAT TACGCGCAGA AAAAAAGGAT CTCAAGAAGG
     3541 CCTACTATTA GCAACAACGA TCCTTTGATC TTTTCTACGG GGTCTGACGC TCAGTGGAAC
     3601 GAAAACTCAC GTTAAGGGAT TTTGGTCATG AGATTATCAA AAAGGATCTT CACCTAGATC
     3661 CTTTTAAATT AAAAATGAAG TTTTAAATCA ATCTAAAGTA TATATGAGTA AACTTGGTCT
     3721 GACAGTTACC AATGCTTAAT CAGTGAGGCA CCTATCTCAG CGATCTGTCT ATTTCGTTCA
     3781 TCCATAGTTG CCTGACTCCC CGTCGTGTAG ATAACTACGA TACGGGAGGG CTTACCATCT
     3841 GGCCCCAGTG CTGCAATGAT ACCGCGAGAA CCACGCTCAC CGGCTCCAGA TTTATCAGCA
     3901 ATAAACCAGC CAGCCGGAAG GGCCGAGCGC AGAAGTGGTC CTGCAACTTT ATCCGCCTCC
     3961 ATCCAGTCTA TTAATTGTTG CCGGGAAGCT AGAGTAAGTA GTTCGCCAGT TAATAGTTTG
     4021 CGCAACGTTG TTGCCATTGC TACAGGCATC GTGGTGTCAC GCTCGTCGTT TGGTATGGCT
     4081 TCATTCAGCT CCGGTTCCCA ACGATCAAGG CGAGTTACAT GATCCCCCAT GTTGTGCAAA
     4141 AAAGCGGTTA GCTCCTTCGG TCCTCCGATC GTTGTCAGAA GTAAGTTGGC CGCAGTGTTA
     4201 TCACTCATGG TTATGGCAGC ACTGCATAAT TCTCTTACTG TCATGCCATC CGTAAGATGC
     4261 TTTTCTGTGA CTGGTGAGTA CTCAACCAAG TCATTCTGAG AATAGTGTAT GCGGCGACCG
     4321 AGTTGCTCTT GCCCGGCGTC AATACGGGAT AATACCGCGC CACATAGCAG AACTTTAAAA
     4381 GTGCTCATCA TTGGAAAACG TTCTTCGGGG CGAAAACTCT CAAGGATCTT ACCGCTGTTG
     4441 AGATCCAGTT CGATGTAACC CACTCGTGCA CCCAACTGAT CTTCAGCATC TTTTACTTTC
     4501 ACCAGCGTTT CTGGGTGAGC AAAAACAGGA AGGCAAAATG CCGCAAAAAA GGGAATAAGG
     4561 GCGACACGGA AATGTTGAAT ACTCATACTC TTCCTTTTTC AATATTATTG AAGCATTTAT
     4621 CAGGGTTATT GTCTCATGAG CGGATACATA TTTGAATGTA TTTAGAAAAA TAAACAAATA
     4681 GGGGTTCCGC GCACCTGCAC CAGTCAGTAA AACGACGGCC AGTAGTCAAA AGCCTCCGAC
     4741 CGGAGGCTTT TGACTTGGTT CAGGTGGAGT GGGAG
//