LOCUS       V015840                 7190 bp    DNA     circular SYN 19-APR-2020
DEFINITION  Exported.
ACCESSION   V015840
VERSION     V015840
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7190)
  AUTHORS   Fischer EC, Hashimoto K, Zhang Y, Feldman AW, Dien VT, Karadeema RJ,
            Adhikary R, Ledbetter MP, Krishnamurthy R, Romesberg FE.
  TITLE     New codons for efficient production of unnatural proteins in a
            semi-synthetic organism
  JOURNAL   Nat. Chem. Biol. (2020) In press
REFERENCE   2  (bases 1 to 7190)
  AUTHORS   Fischer EC.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-DEC-2019) Department of Chemistry, The Scripps
            Research Institute, 10550 North Torrey Pines Road, La Jolla, CA
            92037, USA
REFERENCE   3  (bases 1 to 7190)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem.
            Biol. (2020) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (27-DEC-2019) Department of Chemistry, The Scripps Research
            Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA"
FEATURES             Location/Qualifiers
     source          1..7190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      19..607
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     regulatory      717..786
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     CDS             846..1466
                     /label="TetR"
                     /note="tetracycline repressor TetR"
     terminator      complement(1491..1577)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     promoter        1893..1970
                     /label="lacIq promoter"
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             1971..3050
                     /label="lacI"
                     /note="lac repressor"
     promoter        3171..3201
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    3209..3225
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             3254..4171
                     /gene="tyrS"
                     /label="Tyrosine--tRNA ligase"
                     /note="Tyrosine--tRNA ligase from Methanocaldococcus
                     jannaschii (strain ATCC 43067 / DSM 2661 / JAL-1 / JCM
                     10045 / NBRC 100440). Accession#: Q57834"
     promoter        4371..4399
                     /label="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    4407..4423
                     /label="lacI binding site"
                     /bound_moiety="lacI"
                     /note="lac repressor binds to the lac operator to inhibit
                     transcription in E. coli; lac operator"
     CDS             4446..5705
                     /codon_start=1
                     /transl_table=11
                     /product="chimeric PylRS variant"
                     /label="chimeric PylRS variant"
                     /note="chimeric PylRS variant; Methanosarcina mazei/barkeri
                     chPylRS(IPYE)"
                     /protein_id="QIZ64222.1"
                     /translation="MDKKPLDVLISATGLWMSRTGTLHKIKHYEISRSKIYIEMACGDH
                     LVVNNSRSCRPARAFRYHKYRKTCKRCRVSDEDINNFLTRSTEGKTSVKVKVVSEPKVK
                     KAMPKSVSRAPKPLENPVSAKASTDTSRSVPSPAKSTPNSPVPTSASAPALTKSQTDRL
                     EVLLNPKDEISLNSGKPFRELESELLSRRKKDLQQIYAEERENYLGKLEREITRFFVDR
                     GFLEIKSPILIPLEYIERMGIDNDTELSKQIFRVDKNFCLRPMLAPNLYNYLRKLDRAL
                     PDPIKIFEIGPCYRKESDGKEHLEEFTMLNFCQMGSGCTRENLESIITDFLNHLGIDFK
                     IVGDSCMVYGDTLDVMHGDLELSSAVVGPIPLDREWGIDKPWIGAGFGLERLLKVKHDF
                     KNIKRAARSESYYNGISTNL"
     terminator      5929..5976
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     regulatory      6073..6177
                     /label="AmpR promoter"
                     /note="AmpR promoter"
                     /regulatory_class="promoter"
     CDS             6178..7035
                     /label="AmpR"
                     /note="beta-lactamase"
ORIGIN
        1 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
       61 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
      121 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
      181 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      241 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      301 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      361 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      421 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      481 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      541 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      601 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      661 tcacatgttc tttcctgcgt tatccccgag ctcttagcgc gaattgtcga gggaaatttt
      721 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
      781 taatattaaa tatggctggt tctcgcagaa agaaacatat ccatgaaatc ccgccccgaa
      841 ttgatatgtc cagattagat aaaagtaaag tgattaacag cgcattagag ctgcttaatg
      901 aggtcggaat cgaaggttta acaacccgta aactcgccca gaagctaggt gtagagcagc
      961 ctacattgta ttggcatgta aaaaataagc gggctttgct cgacgcctta gccattgaga
     1021 tgttagatag gcaccatact cacttttgcc ctttagaagg ggaaagctgg caagattttt
     1081 tacgtaataa cgctaaaagt tttagatgtg ctttactaag tcatcgcgat ggagcaaaag
     1141 tacatttagg tacacggcct acagaaaaac agtatgaaac tctcgaaaat caattagcct
     1201 ttttatgcca acaaggtttt tcactagaga atgcattata tgcactcagc gctgtggggc
     1261 attttacttt aggttgcgta ttggaagatc aagagcatca agtcgctaaa gaagaaaggg
     1321 aaacacctac tactgatagt atgccgccat tattacgaca agctatcgaa ttatttgatc
     1381 accaaggtgc agagccagcc ttcttattcg gccttgaatt gatcatttgc ggattagaaa
     1441 aacaacttaa atgtgaaagt gggtcttaag cactaggtct agggcggcgg atttgtccta
     1501 ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc tttcgactga
     1561 gcctttcgtt ttatttgatg cctctagcac gcgtagagct agagccttca acccagtcag
     1621 ctccttccgg tgggcgcggg gcatgactaa catgagaatt acaacttata tcgtatgggg
     1681 ctgacttcag gtgctacatt tgaagagata aattgcactg aaatctagat gattctgtgg
     1741 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     1801 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     1861 atctgtgcgg tatttcacac cgcataaatt ccgacaccat cgaatggtgc aaaacctttc
     1921 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
     1981 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
     2041 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
     2101 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
     2161 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
     2221 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
     2281 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
     2341 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
     2401 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
     2461 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
     2521 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
     2581 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
     2641 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
     2701 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
     2761 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
     2821 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
     2881 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
     2941 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     3001 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     3061 gcaatgtaaa tcactgcata attcgtgtcg ctcaaggcgc actcccgttc tggataatgt
     3121 tttttgcgcc gacatcataa cggttctggc aaatattagg caccccaggc tttacacttt
     3181 atgcttccgg ctggtataat gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     3241 agtattcaga tctatggacg agttcgaaat gattaaacgc aacaccagcg aaattatctc
     3301 tgaagaagag ctgcgcgagg tgctgaagaa agacgagaag agcgcgacta ttggctttga
     3361 gccgtccggt aaaattcacc tgggtcacta cctgcaaatc aagaagatga ttgatctgca
     3421 aaacgctggt tttgacatca ttatcctgct ggcggacctg cacgcctacc tgaatcaaaa
     3481 gggcgagctg gatgagattc gcaagatcgg cgactacaat aagaaagtct tcgaagccat
     3541 gggtttgaag gctaaatacg tctacggtag caattttcag ctggataagg attacacgtt
     3601 gaatgtgtac cgtctggcgc tgaaaaccac gctgaaacgc gcccgtcgtt ccatggagct
     3661 gattgcgcgc gaggatgaga atccaaaagt tgctgaggtt atttacccta ttatgcaagt
     3721 taatccgttg cactaccagg gtgttgatgt tgccgtcggt ggtatggagc aacgcaaaat
     3781 tcacatgctg gcacgtgaac tgctgccgaa aaaggttgtc tgtattcata atccggtcct
     3841 gaccggcctg gatggcgagg gtaaaatgag cagcagcaag ggtaacttta ttgcagttga
     3901 cgatagcccg gaagaaatcc gtgcgaagat caagaaagcg tactgcccgg caggcgtggt
     3961 tgagggtaac ccgatcatgg aaatcgccaa gtattttctg gaatacccac tgacgattaa
     4021 gcgcccggag aaatttggcg gcgacctgac cgtcaacagc tacgaggagc tggaaagctt
     4081 gtttaagaac aaagaactgc atccgatgcg cctgaaaaac gccgtggcgg aagagctgat
     4141 taagattctg gaaccaattc gcaaacgtct gtaagtcgac tgatgactga cgatctgcct
     4201 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     4261 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     4321 tggcggcgcc gacatcataa cggttctggc aaatattctg aaatgagctg ttgacaatta
     4381 atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca caggaaacag
     4441 tattcatgga taaaaaaccg ctggacgttc tgatctccgc tacgggtctg tggatgagcc
     4501 gcacgggtac gctgcataaa atcaagcact atgagatttc tcgttctaaa atctacatcg
     4561 aaatggcgtg tggtgaccat ctggttgtga acaactctcg ttcttgtcgt ccggcacgtg
     4621 cattccgtta tcataaatac cgtaaaacct gcaaacgttg tcgtgtttct gacgaagata
     4681 tcaacaactt cctgacccgt tctaccgaag gcaaaacctc tgttaaagtt aaagttgttt
     4741 ctgaaccgaa agtgaaaaaa gcgatgccga aatctgtttc tcgtgcgccg aaaccgctgg
     4801 aaaatccggt ttctgcgaaa gcgtctaccg acacctctcg ttctgttccg tctccggcga
     4861 aatctacccc gaactctccg gttccgacct ctgcaagtgc ccccgcactt acgaagagcc
     4921 agactgacag gcttgaagtc ctgttaaacc caaaagatga gatttccctg aattccggca
     4981 agcctttcag ggagcttgag tccgaattgc tctctcgcag aaaaaaagac ctgcagcaga
     5041 tctacgcgga agaaagggag aattatctgg ggaaactcga gcgtgaaatt accaggttct
     5101 ttgtggacag gggttttctg gaaataaaat ccccgatcct gatccctctt gagtatatcg
     5161 aaaggatggg cattgataat gataccgaac tttcaaaaca gatcttcagg gttgacaaga
     5221 acttctgcct gagacccatg cttgctccaa acctttacaa ctacctgcgc aagcttgaca
     5281 gggccctgcc tgatccaata aaaatttttg aaataggccc atgctacaga aaagagtccg
     5341 acggcaaaga acacctcgaa gagtttacca tgctgaactt ctgccagatg ggatcgggat
     5401 gcacacggga aaatcttgaa agcataatta cggacttcct gaaccacctg ggaattgatt
     5461 tcaagatcgt aggcgattcc tgcatggtct atggggatac ccttgatgta atgcacggag
     5521 acctggaact ttcctctgca gtagtcggac ccataccgct tgaccgggaa tggggtattg
     5581 ataaaccctg gataggggca ggtttcggac tcgaacgcct tctaaaggtt aaacacgact
     5641 ttaaaaatat caagagagct gcacgctcgg aatcgtatta caacggcatc tcaaccaatc
     5701 tgtaatagtg actgacgatc tgcctcgcgc gtttcggtga tgacggtgaa aacctaggtt
     5761 ttgtcgacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     5821 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
     5881 ccatgaccca gtcacgtagc gatagcggag tgggatcctt ttgcatgcct agcataaccc
     5941 cttggggcct ctaaacgggt cttgaggggt tttttgagac gaaagggcct cgtgatacgc
     6001 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     6061 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6121 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     6181 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     6241 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     6301 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6361 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6421 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6481 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6541 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6601 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6661 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6721 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6781 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6841 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6901 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6961 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     7021 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     7081 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7141 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
//
