LOCUS       Exported                4682 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expression plasmid encoding the evolved MjTyrRS variant Iodo.5RS for
            amber suppression with p-iodophenylalanine.
ACCESSION   .
VERSION     .
KEYWORDS    pDB070.iodo.5
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4682)
  AUTHORS   Bryson DI, Fan C, Guo LT, Miller C, Soll D, Liu DR
  TITLE     Continuous directed evolution of aminoacyl-tRNA synthetases.
  JOURNAL   Nat Chem Biol. 2017 Oct 16. doi: 10.1038/nchembio.2474.
  PUBMED    29035361
REFERENCE   2  (bases 1 to 4682)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4682
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(138..155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(272..817)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(352..371)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(965..1588)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     1496..1515
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    1696..1714
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        1715..1769
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    1720..1738
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    1750..1768
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(2740..2757)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2910..2956
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3666..4325)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4240..4259
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4326..4428)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 atcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg gtgtcagtga
       61 agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac
      121 aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga
      181 gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac
      241 agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc
      301 atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc
      361 aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg
      421 gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg
      481 taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc
      541 gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg
      601 gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg
      661 ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa
      721 agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc
      781 agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat
      841 aaaatatttc tagatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc
      901 catacgatat aagttgtaat tctcatgttt gacagcttat catcgataaa gctatctaga
      961 aagcttaaga cccactttca catttaagtt gtttttctaa tccgcatatg atcaattcaa
     1021 ggccgaataa gaaggctggc tctgcacctt ggtgatcaaa taattcgata gcttgtcgta
     1081 ataatggcgg catactatca gtagtaggtg tttccctttc ttctttagcg acttgatgct
     1141 cttgatcttc caatacgcaa cctaaagtaa aatgccccac tgcgctgagt gcatataatg
     1201 cattctctag tgaaaaacct tgttggcata aaaaggctaa ttgattttcg agagtttcat
     1261 actgtttttc tgtaggccgt gtacctaaat gtacttttgc tccatcgcga tgacttagta
     1321 aagcacatct aaaactttta gcgttattac gtaaaaaatc ttgccagctt tccccttcta
     1381 aagggcaaaa gtgagtatgg tgcctatcta acatctcaat ggctaaggcg tcgagcaaag
     1441 cccgcttatt ttttacatgc caatacaatg taggctgctc tacacctagc ttctgggcga
     1501 gtttacgggt tgttaaacct tcgattccga cctcattaag cagctctaat gcgctgttaa
     1561 tcactttact tttatctaat ctagacatat gaattcgggg ctttatgaat ctattataca
     1621 gaaaaatttt cctgaaagca aataattaat tgcgttgcgc attctagagc acagctccat
     1681 cgaatggcca gatgatccct atcagtgata gagattgaca ctctatcatt gatagagtta
     1741 ttttaccact ccctatcagt gatagagaaa gacctatgga cgaatttgaa atgataaaga
     1801 gaaacacatc tgaaattatc agcgaggaag agttaagaga ggttttaaaa aaagatgaaa
     1861 aatctgctct gataggtttt gaaccaagtg gtaaaataca tttagggcat tatctccaaa
     1921 taaaaaagat gattgattta caaaatgctg gatttgatat aattatattg ttggctgatt
     1981 tccacgccta tttaaaccag aaaggagagt tggatgagat tagaaaaata ggagattata
     2041 acaaaaaagt ttttgaagca atggggttaa aggcaaaata tgtttatgga agttcgttcc
     2101 agcttgataa ggattataca ctgaatgtct atagattggc tttaaaaact accttaaaaa
     2161 gagcaagaag gagtatggaa cttatagcaa gagaggatga aaatccaaag gttgctgaag
     2221 ttatctatcc aataatgcag gttaatcctc ttaattatga gggcgttgat gttgcagttg
     2281 gagggatgga gcagagaaaa atacacatgt tagcaaggga gcttttacca aaaaaggttg
     2341 tttgtattca caaccctgtc ttaacgggtt tggatggaga aggaaagatg agttcttcaa
     2401 aagggaattt tatagctgtt gatgactctc cagaagagat tagggctaag ataaagaaag
     2461 catactgccc agctggaatt gttgaaggaa atccaataat ggagatagct aaatacttcc
     2521 ttgaatatcc tttaaccata aaaaggccag aaaaatttgg tggagatttg acagttaata
     2581 gctatgagga gttagagagt ttatttaaaa ataaggaatt gcatccaatg cgcttaaaaa
     2641 atgctgtagc tgaagaactt ataaagattt tagagccaat tagaaagaga ttataagtct
     2701 ccagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga
     2761 acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc
     2821 tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc
     2881 ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     2941 gggcctttcg ttttatctgt tgtttgtgag ctcccggtca tcaatcatcc ccataatcct
     3001 tgttagatta tcaaagtttc aaacgctaaa ttgcctgatg cgctacgctt atcaggccta
     3061 catgatctct gcaatatatt gagtttgcgt gcttttgtag gccggataag gcgttcacgc
     3121 cgcatccggc aagaaacagc aaacaatcca aaacgccgcg ttcagcggcg ttttttctgc
     3181 ttttcttcgc gaattaattc cgcttcgcaa catgtgagca ccggtttatt gactaccgga
     3241 agcagtgtga ccgtgtgctt ctcaaatgcc tgaggccagt ttgctcaggc tctccccgtg
     3301 gaggtaataa ttgacgatat gatcagtgca cggctaacta agcggcctgc tgactttctc
     3361 gccgatcaaa aggcattttg ctattaaggg attgacgagg gcgtatctgc gcagtaagat
     3421 gcgccccgca ttccggcggt agttcagcag ggcagaacgg cggactctaa atccgcatgg
     3481 caggggttca aatcccctcc gccggaccaa attcgaaaag cctgctcaac gagcaggctt
     3541 ttttgcatgc tcgagcagct cagggtcgaa tttgctttcg aatttctgcc attcatccgc
     3601 ttattatcac ttattcaggc gtagcaacca ggcgtttaag ggcaccaata actgccttaa
     3661 aaaaattacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg
     3721 ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc
     3781 ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata
     3841 ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac
     3901 atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct
     3961 tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa
     4021 aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc
     4081 agctcaccgt ctttcattgc catacggaat tccggatgag cattcatcag gcgggcaaga
     4141 atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc
     4201 gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca
     4261 aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc
     4321 tccattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt
     4381 gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaac gtctcatttt
     4441 cgccaaaagt tggcccaggg cttcccggta tcaacaggga caccaggatt tatttattct
     4501 gcgaagtgat cttccgtcac aggtatttat tcggcgcaaa gtgcgtcggg tgatgctgcc
     4561 aacttactga tttagtgtat gatggtgttt ttgaggtgct ccagtggctt ctgtttctat
     4621 cagctgtccc tcctgttcag ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg
     4681 ac
//
