LOCUS       Exported     4451 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..4451
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
 polymerase"
     terminator      2488..2535
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA polymerase"
     primer_bind     4383..4399
                     /label="M13 for"
                     /note="common sequencing primer, one of multiple similar variants"
     terminator      4400..4431
                     /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     2371..2390
                     /label="dialout_rev"
                     /note="Forward dialout primer. Unique pair for each FreeGenes part."
     CDS             29..2356
                     /label="KOD DNA polymerase"
                     /note=""
                     /translation="MILDTDYITEDGKPVIRIFKKENGEFKIEYDRTFEPYFYALLKDDSAIEEVKKITAERHGTVVTVKRVEKVQKKFLGRPVEVWKLYFTHPQDVPAIRDKIREHPAVIDIYEYDIPFAKRYLIDKGLVPMEGDEELKMLAFDIETLYHEGEEFAEGPILMISYADEEGARVITWKNVDLPYVDVVSTEREMIKRFLRVVKEKDPDVLITYNGDNFDFAYLKKRCEKLGINFALGRDGSEPKIQRMGDRFAVEVKGRIHFDLYPVIRRTINLPTYTLEAVYEAVFGQPKEKVYAEEITTAWETGENLERVARYSMEDAKVTYELGKEFLPMEAQLSRLIGQSLWDVSRSSTGNLVEWFLLRKAYERNELAPNKPDEKELARRRQSYEGGYVKEPERGLWENIVYLDFRCSLYPSIIITHNVSPDTLNREGCKEYDVAPQVGHRFCKDFPGFIPSLLGDLLEERQKIKKKMKATIDPIERKLLDYRQRAIKILANSYYGYYGYARARWYCKECAESVTAWGREYITMTIKEIEEKYGFKVIYSDTDGFFATIPGADAETVKKKAMEFLKYINAKLPGALELEYEGFYKRGFFVTKKKYAVIDEEGKITTRGLEIVRRDWSEIAKETQARVLEALLKDGDVEKAVRIVKEVTEKLSKYEVPPEKLVIHEQITRDLKDYKATGPHVAVAKRLAARGVKIRPGTVISYIVLKGSGRIGDRAIPFDEFDPTKHKYDAEYYIENQVLPAVERILRAFGYRKEDLRYQKTRQVGLSAWLKPKGT*"
     primer_bind     complement(2540..2556)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar variants"
     rep_origin      complement(2623..3211)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
     CDS             complement(3402..4262)
                     /label="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"
     promoter        complement(4263..4367)
                     /label="bla promoter"
                     /note="ampicillin resistance gene promoter"
     misc_feature    2391..2450
                     /label="FreeGenes Barcode"
                     /note="FreeGenes Barcode. Defined as I-SceI site flanked by 21 base pair random sequences."
ORIGIN
        1 AAAGGGGCCG TCAATATCAG GGTCTCTAAT GATCCTGGAC ACCGATTACA TCACCGAGGA
       61 TGGCAAACCG GTGATCCGTA TCTTCAAAAA AGAAAATGGC GAGTTCAAAA TTGAATACGA
      121 CCGCACCTTT GAACCGTATT TCTATGCTCT GCTGAAAGAT GATTCCGCCA TTGAAGAGGT
      181 TAAAAAAATT ACTGCTGAAC GCCATGGCAC AGTTGTTACC GTTAAACGCG TCGAAAAGGT
      241 CCAAAAAAAG TTCCTGGGTC GCCCGGTAGA AGTTTGGAAA CTGTATTTTA CCCACCCGCA
      301 AGACGTACCG GCTATCCGCG ACAAAATCCG CGAGCACCCA GCGGTGATCG ATATCTACGA
      361 GTATGATATC CCGTTTGCGA AACGCTACCT GATCGACAAG GGTCTGGTGC CGATGGAAGG
      421 CGACGAGGAG CTGAAAATGC TGGCATTCGA CATCGAAACA CTGTATCACG AAGGCGAAGA
      481 GTTCGCTGAA GGCCCGATCC TGATGATCTC GTACGCTGAC GAAGAAGGCG CCCGTGTTAT
      541 CACATGGAAA AACGTGGATT TGCCGTATGT TGACGTGGTA TCCACCGAAC GTGAAATGAT
      601 CAAGCGTTTC CTGCGAGTGG TTAAAGAGAA AGACCCGGAC GTTCTGATCA CCTACAACGG
      661 CGACAACTTC GATTTTGCCT ACCTGAAAAA ACGCTGCGAA AAACTCGGTA TCAACTTCGC
      721 CCTGGGTCGT GACGGTTCGG AACCTAAAAT CCAGCGTATG GGTGACCGTT TCGCTGTTGA
      781 AGTAAAGGGC CGCATCCATT TCGACCTGTA TCCAGTTATC CGTCGTACAA TCAACCTGCC
      841 AACCTACACT CTGGAAGCGG TATACGAGGC GGTTTTCGGC CAACCGAAAG AAAAAGTTTA
      901 CGCTGAAGAA ATCACCACTG CATGGGAAAC CGGCGAAAAT CTGGAACGTG TAGCGCGCTA
      961 CAGCATGGAG GACGCGAAAG TTACTTACGA ACTGGGAAAA GAGTTCCTCC CGATGGAAGC
     1021 ACAGCTGAGC CGTCTTATTG GCCAGTCTCT GTGGGACGTT TCCCGTTCTT CTACCGGTAA
     1081 CCTGGTTGAG TGGTTCCTGC TGCGCAAAGC TTATGAGCGT AACGAACTGG CACCGAATAA
     1141 ACCAGATGAA AAAGAACTGG CACGTCGTCG CCAATCTTAT GAGGGTGGGT ATGTGAAAGA
     1201 ACCGGAACGT GGTCTGTGGG AAAACATCGT CTATCTGGAC TTCCGTTGCA GCCTGTACCC
     1261 AAGCATTATC ATCACTCACA ACGTGTCACC GGACACTCTG AACCGTGAAG GCTGTAAGGA
     1321 ATATGATGTT GCGCCGCAGG TAGGTCACCG TTTTTGCAAA GACTTCCCGG GTTTTATCCC
     1381 GAGCCTGCTG GGTGATCTTC TTGAAGAACG CCAGAAAATT AAAAAGAAAA TGAAGGCCAC
     1441 CATCGACCCG ATTGAACGCA AACTGCTGGA TTATCGCCAG CGCGCTATTA AGATTCTGGC
     1501 TAATTCTTAC TATGGCTACT ACGGCTACGC TCGCGCACGC TGGTACTGCA AAGAGTGTGC
     1561 TGAATCCGTA ACCGCTTGGG GCCGTGAATA TATCACAATG ACCATTAAAG AAATCGAGGA
     1621 GAAATACGGT TTCAAGGTTA TTTATAGCGA TACTGACGGC TTCTTTGCGA CCATCCCAGG
     1681 CGCGGACGCA GAAACCGTAA AGAAGAAAGC AATGGAGTTT CTTAAATATA TTAACGCTAA
     1741 ATTGCCGGGC GCGCTGGAGC TGGAATACGA GGGTTTCTAC AAGCGTGGGT TCTTCGTGAC
     1801 GAAGAAGAAG TACGCAGTAA TCGACGAAGA AGGCAAAATT ACCACTCGCG GCTTGGAAAT
     1861 CGTTCGCCGT GACTGGTCCG AAATTGCTAA AGAAACCCAG GCTCGTGTAC TGGAGGCCTT
     1921 GCTGAAAGAT GGCGACGTAG AAAAAGCGGT TCGTATCGTG AAAGAAGTAA CCGAAAAGCT
     1981 GTCAAAATAC GAAGTTCCGC CAGAGAAACT GGTTATCCAC GAACAGATCA CTCGTGATCT
     2041 GAAAGACTAC AAGGCGACGG GTCCGCATGT TGCAGTAGCC AAACGTCTGG CGGCACGTGG
     2101 TGTGAAAATC CGCCCGGGCA CCGTTATCAG TTACATCGTT CTGAAAGGTT CTGGTCGCAT
     2161 CGGTGATCGT GCGATCCCGT TCGATGAGTT CGACCCGACC AAGCACAAAT ACGACGCAGA
     2221 ATACTACATT GAGAACCAGG TGCTTCCGGC GGTCGAACGT ATCCTGCGCG CGTTCGGTTA
     2281 CCGTAAGGAG GACCTGCGTT ACCAGAAAAC TCGCCAGGTA GGCCTGTCCG CATGGCTGAA
     2341 ACCGAAAGGC ACCTGAAGAG CTTTGAGACC ACCACAGGTC AAGATTCACG TTACCCACGC
     2401 CTTCCTTTAG GTAGGGATAA CAGGGTAATT GACAAATCTT CTTGGATGCA CGCTGAGGTG
     2461 TCAATCGTCG GAGCCGCTGA GCAATAACTA GCATAACCCC TTGGGGCCTC TAAACGGGTC
     2521 TTGAGGGGTT TTTTGCATGG TCATAGCTGT TTCCTGAGAG CTTGGCAGGT GATGACACAC
     2581 ATTAACAAAT TTCGTGAGGA GTCTCCAGAA GAATGCCATT AATTTCCATA GGCTCCGCCC
     2641 CCCTGACGAG CATCACAAAA ATCGACGCTC AAGTCAGAGG TGGCGAAACC CGACAGGACT
     2701 ATAAAGATAC CAGGCGTTTC CCCCTGGAAG CTCCCTCGTG CGCTCTCCTG TTCCGACCCT
     2761 GCCGCTTACC GGATACCTGT CCGCCTTTCT CCCTTCGGGA AGCGTGGCGC TTTCTCATAG
     2821 CTCACGCTGT AGGTATCTCA GTTCGGTGTA GGTCGTTCGC TCCAAGCTGG GCTGTGTGCA
     2881 CGAACCCCCC GTTCAGCCCG ACCGCTGCGC CTTATCCGGT AACTATCGTC TTGAGTCCAA
     2941 CCCGGTAAGA CACGACTTAT CGCCACTGGC AGCAGCCACT GGTAACAGGA TTAGCAGAGC
     3001 GAGGTATGTA GGCGGTGCTA CAGAGTTCTT GAAGTGGTGG CCTAACTACG GCTACACTAG
     3061 AAGAACAGTA TTTGGTATCT GCGCTCTGCT GAAGCCAGTT ACCTTCGGAA AAAGAGTTGG
     3121 TAGCTCTTGA TCCGGCAAAC AAACCACCGC TGGTAGCGGT GGTTTTTTTG TTTGCAAGCA
     3181 GCAGATTACG CGCAGAAAAA AAGGATCTCA AGAAGGCCTA CTATTAGCAA CAACGATCCT
     3241 TTGATCTTTT CTACGGGGTC TGACGCTCAG TGGAACGAAA ACTCACGTTA AGGGATTTTG
     3301 GTCATGAGAT TATCAAAAAG GATCTTCACC TAGATCCTTT TAAATTAAAA ATGAAGTTTT
     3361 AAATCAATCT AAAGTATATA TGAGTAAACT TGGTCTGACA GTTACCAATG CTTAATCAGT
     3421 GAGGCACCTA TCTCAGCGAT CTGTCTATTT CGTTCATCCA TAGTTGCCTG ACTCCCCGTC
     3481 GTGTAGATAA CTACGATACG GGAGGGCTTA CCATCTGGCC CCAGTGCTGC AATGATACCG
     3541 CGAGAACCAC GCTCACCGGC TCCAGATTTA TCAGCAATAA ACCAGCCAGC CGGAAGGGCC
     3601 GAGCGCAGAA GTGGTCCTGC AACTTTATCC GCCTCCATCC AGTCTATTAA TTGTTGCCGG
     3661 GAAGCTAGAG TAAGTAGTTC GCCAGTTAAT AGTTTGCGCA ACGTTGTTGC CATTGCTACA
     3721 GGCATCGTGG TGTCACGCTC GTCGTTTGGT ATGGCTTCAT TCAGCTCCGG TTCCCAACGA
     3781 TCAAGGCGAG TTACATGATC CCCCATGTTG TGCAAAAAAG CGGTTAGCTC CTTCGGTCCT
     3841 CCGATCGTTG TCAGAAGTAA GTTGGCCGCA GTGTTATCAC TCATGGTTAT GGCAGCACTG
     3901 CATAATTCTC TTACTGTCAT GCCATCCGTA AGATGCTTTT CTGTGACTGG TGAGTACTCA
     3961 ACCAAGTCAT TCTGAGAATA GTGTATGCGG CGACCGAGTT GCTCTTGCCC GGCGTCAATA
     4021 CGGGATAATA CCGCGCCACA TAGCAGAACT TTAAAAGTGC TCATCATTGG AAAACGTTCT
     4081 TCGGGGCGAA AACTCTCAAG GATCTTACCG CTGTTGAGAT CCAGTTCGAT GTAACCCACT
     4141 CGTGCACCCA ACTGATCTTC AGCATCTTTT ACTTTCACCA GCGTTTCTGG GTGAGCAAAA
     4201 ACAGGAAGGC AAAATGCCGC AAAAAAGGGA ATAAGGGCGA CACGGAAATG TTGAATACTC
     4261 ATACTCTTCC TTTTTCAATA TTATTGAAGC ATTTATCAGG GTTATTGTCT CATGAGCGGA
     4321 TACATATTTG AATGTATTTA GAAAAATAAA CAAATAGGGG TTCCGCGCAC CTGCACCAGT
     4381 CAGTAAAACG ACGGCCAGTA GTCAAAAGCC TCCGACCGGA GGCTTTTGAC TTGGTTCAGG
     4441 TGGAGTGGGA G
//