LOCUS       62056_3535        5741 bp DNA     circular SYN 12-JUN-2021
DEFINITION  Cloning vector pBAD_GST-MBP_wt, complete sequence.
ACCESSION   MW879734
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5741)
  AUTHORS   Robertson WE, Funke LFH., de la Torre D, Fredens J, Elliott TS, 
            Spinck M, Christova Y, Cervettini D, Boge FL, Liu KC, Buse S, Maslen
            S, Salmond GPC., Chin JW.
  TITLE     Sense Codon Reassignment Enables Viral Resistance and Encoded 
            Polymer Synthesis
  JOURNAL   Science (2021) In press
REFERENCE   2  (bases 1 to 5741)
  AUTHORS   Robertson WE, Funke LFH., de la Torre D, Fredens J, Elliott TS, 
            Spinck M, Christova Y, Cervettini D, Boge FL, Liu KC, Buse S, Maslen
            S, Salmond GPC., Chin JW.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-APR-2021) Protein and Nucleic Acid Chemistry, MRC 
            Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, 
            Cambridgeshire CB2 0QH, United Kingdom
REFERENCE   3  (bases 1 to 5741)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science 
            (2021) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-APR-2021) Protein and Nucleic Acid Chemistry, MRC Laboratory of 
            Molecular Biology, Francis Crick Avenue, Cambridge, Cambridgeshire 
            CB2 0QH, United Kingdom"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5741
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(266..811)
                     /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."
     promoter        923..951
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     promoter        999..1090
                     /label=AmpR promoter
     CDS             1091..1891
                     /codon_start=1
                     /label=ApmR
                     /note="aminoglycoside 3-N-acetyltransferase type IV"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     CDS             complement(2205..3080)
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3107..3391
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     CDS             3425..4078
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             4088..4108
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQS"
     CDS             4127..4144
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     CDS             4157..5257
                     /codon_start=1
                     /label=MBP
                     /note="maltose binding protein from E. coli"
                     /translation="LKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     CDS             5337..5354
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      5580..5666
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct
       61 tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata
      121 tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa
      181 atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa
      241 gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg
      301 aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt
      361 ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca
      421 ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca
      481 gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta
      541 tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca
      601 gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac
      661 tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt
      721 ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca
      781 agagattacg cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata
      841 tttctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg
      901 atataagttg taattctcat gtttgacagc ttatcatcga taagctttaa tgcggtagtt
      961 tatcacagtt aaattgctaa cgcagtcagg caccgtgttt tgtttatttt tctaaataca
     1021 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     1081 aaggaagagt atgagtagtg cggtggagtg caatgtcgtg caatacgaat ggcgaaaagc
     1141 cgagctcatc ggtcagcttc tcaaccttgg ggttaccccc ggcggtgtgc tgctggtcca
     1201 cagctccttc cgtagcgtcc ggcccctcga agatgggcca cttggactga tcgaggccct
     1261 gcgtgctgcg ctgggtccgg gagggacgct cgtcatgccc agctggagtg gtctggacga
     1321 cgagccgttc gatcctgcca cgagccccgt tacaccggac cttggagttg tctctgacac
     1381 attctggcgc ctgccaaatg taaagcgcag cgcccatcca tttgcctttg cggcagcggg
     1441 gccacaggca gagcagatca tctctgatcc attgcccctg ccacctcaca gccctgcaag
     1501 cccggtcgcc cgtgtccatg aactcgatgg gcaggtactt ctcctcggcg tgggacacga
     1561 tgccaacacg acgctgcatc ttgccgagtt gatggcaaag gttccctatg gggtgccgag
     1621 acactgcacc attcttcagg atggcaagtt ggtacgcgtc gattatctcg agaatgacca
     1681 ctgctgtgag cgctttgcct tggcggacag gtggctcaag gagaagagcc ttcagaagga
     1741 aggtccagtc ggtcatgcct ttgctcggtt gatccgctcc cgcgacattg tggcgacagc
     1801 cctgggtcaa ctgggccgag atccgttgat cttcctgcat ccgccagagg cgggatgcga
     1861 agaatgcgat gccgctcgcc agagcattgg ctgaatggaa gccggcggca cctcgctaac
     1921 ggattcacca ctccaagaat tggagccaat caattcttgc ggagaactgt gaatgcgcaa
     1981 accaaccctt ggcagaacat atccatcgcg tgcggccgcg tctccgggag ctgcatgtgt
     2041 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag
     2101 gcgaagcggc atgcataatg tgcctgtcaa atggacgaag cagggattct gcaaacccta
     2161 tgctactccg tcaagccgtc aattgtctga ttcgttacca attaactcaa cttgacggct
     2221 acatcattca ctttttcttc acaaccggca cggaactcgc tcgggctggc cccggtgcat
     2281 tttttaaata cccggctgaa atagagttga tcgtcaaaac caacattgcg accgacggtg
     2341 gcgataggca tccgggtggt gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg
     2401 cgccagctta agacgctaat ccctaactgc tggcggaaaa gatgactcag acggctcggg
     2461 ctcaagcaaa catgctgtgc gacgctggcg atatcaaaat tgctgtctgc caggtgatcg
     2521 ctgatgtact gacaagcctc gcgtacccga ttatccatcg gtggatggag gctctcgtta
     2581 atcgcttcca tgcgccgcag taacaattgc tcaagcagat ttatcgccag cagctcgcta
     2641 tagcgccctt ccccttgccc ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc
     2701 tggtgcgctt catccgggcg aaagaacccc gtattggcaa atatactcgg ccagttaagc
     2761 cattcatgcc agtaggcgcg cggacgaaag taaacccact ggtgatacca ttcgcgagcc
     2821 tccggatgac gaccgtagtg atgaatctct cctggcggga acagcaaaat atcacccggt
     2881 cggcaaacaa attctcgtcc ctgatttttc accaccccct gaccgcgaat ggtgagattg
     2941 agaatataac ctttcattcc cagcggtcgg tcgataaaaa aatcgagata accgttggcc
     3001 tcaatcggcg ttaaacccgc caccagatgg gcattaaagc tgtatcccgg cagcagggga
     3061 tcattttgcg cttcagccat acttttcata ctcccgccat tcagagaaga aaccaattgt
     3121 ccatattgca tcagacattg ccgtcactgc gtcttttact ggctcttctc gctaaccaaa
     3181 ccggtaaccc cgcttattaa aagcattctg taacaaagcg ggaccaaagc catgacaaaa
     3241 acgcgtaaca aaagtgtcta taatcacggc agaaaagtcc acattgatta tttgcacggc
     3301 gtcacacttt gctatgccat agcattttta tccataagat tagcggatcc tacctgacgc
     3361 tttttatcgc aactctctac tgtttctcca tacccgtttt ttgggctaac aggaggaatt
     3421 aaccatgtcc cctatactag gttattggaa aattaagggc cttgtgcaac ccactcgact
     3481 tcttttggaa tatcttgaag aaaaatatga agagcatttg tatgagcgcg atgaaggtga
     3541 taaatggcga aacaaaaagt ttgaattggg tttggagttt cccaatcttc cttattatat
     3601 tgatggtgat gttaaattaa cacagtctat ggccatcata cgttatatag ctgacaagca
     3661 caacatgttg ggtggttgtc caaaagagcg tgcagagatt agtatgcttg aaggagcggt
     3721 tttggatatt agatacggtg ttagcagaat tgcatatagt aaagactttg aaactctcaa
     3781 agttgatttt cttagcaagc tacctgaaat gctgaaaatg ttcgaagatc gtttatgtca
     3841 taaaacatat ttaaatggtg atcatgtaac ccatcctgac ttcatgttgt atgacgctct
     3901 tgatgttgtt ttatacatgg acccaatgtg cctggatgcg ttcccaaaat tagtttgttt
     3961 taaaaaacgt attgaagcta tcccacaaat tgataagtac ttgaaatcca gcaagtatat
     4021 agcatggcct ttgcagggct ggcaagccac gtttggtggt ggcgaccatc ctccaaaatc
     4081 tgatagtgaa aacctgtact tccaaagcgg aattcatcgt gacgggctgg ttccgcgtgg
     4141 atccccgggg cagtacctca aaatcgaaga aggtaaactg gtaatctgga ttaacggcga
     4201 taaaggctat aacggtctcg ctgaagtcgg taagaaattc gagaaagata ccggaattaa
     4261 agtcaccgtt gagcatccgg ataaactgga agagaaattc ccacaggttg cggcaactgg
     4321 cgatggccct gacattatct tctgggcaca cgaccgcttt ggtggctacg ctcaatctgg
     4381 cctgttggct gaaatcaccc cggacaaagc gttccaggac aagctgtatc cgtttacctg
     4441 ggatgccgta cgttacaacg gcaagctgat tgcttacccg atcgctgttg aagcgttaag
     4501 cctgatttat aacaaagatc tgctgccgaa cccgccaaaa acctgggaag agatcccggc
     4561 gctggataaa gaactgaaag cgaaaggtaa gagcgcgctg atgttcaacc tgcaagaacc
     4621 gtacttcacc tggccgctga ttgctgctga cgggggttat gcgttcaagt atgaaaacgg
     4681 caagtacgac attaaagacg tgggcgtgga taacgctggc gcgaaagcgg gtctgacctt
     4741 cctggttgac ctgattaaaa acaaacacat gaatgcagac accgattact ccatcgcaga
     4801 agctgccttt aataaaggcg aaacagcgat gaccatcaac ggcccgtggg catggtccaa
     4861 catcgacacc agcaaagtga attatggtgt aacggtactg ccgaccttca agggtcaacc
     4921 atccaaaccg ttcgttggcg tgctgagcgc aggtattaac gccgccagtc cgaacaaaga
     4981 gctggcaaaa gagttcctcg aaaactatct gctgactgat gaaggtctgg aagcggttaa
     5041 taaagacaaa ccgctgggtg ccgtagcgct gaagtcttac gaggaagagt tggcgaaaga
     5101 tccacgtatt gccgccacta tggaaaacgc ccagaaaggt gaaatcatgc cgaacatccc
     5161 gcagatgtcc gctttctggt atgccgtgcg tactgcggtg atcaacgccg ccagcggtcg
     5221 tcagactgtc gatgaagccc tgaaagacgc gcagactagg attacgaaat aaagctcgag
     5281 cgaagcttgg gcccgaacaa aaaatctcag aagaggatct gaatagcgcc gtcgaccatc
     5341 atcatcatca tcattgagtt taaacggtct ccagcttggc tgttttggcg gatgagagaa
     5401 gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt
     5461 gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg
     5521 ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc
     5581 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     5641 tgaacgctct cctgagtagg acaaatccgc cgggagctgt ccctcctgtt cagctactga
     5701 cggggtggtg cgtaacggca aaagcaccgc cggacatcag c
//
