LOCUS       MW879723                4347 bp    DNA     circular SYN 12-JUN-2021
DEFINITION  Cloning vector pMB1_MmPylRS-pylT-opt-serT.UGA, complete sequence.
ACCESSION   MW879723
VERSION     MW879723.1  GI:2049107077
KEYWORDS    .
SOURCE      Cloning vector pMB1_MmPylRS-pylT-opt-serT.UGA
  ORGANISM  Cloning vector pMB1_MmPylRS-pylT-opt-serT.UGA
            other sequences; artificial sequences; vectors.
REFERENCE   1  (bases 1 to 4347)
  AUTHORS   Robertson,W.E., Funke,L.F.H., de la Torre,D., Fredens,J.,
            Elliott,T.S., Spinck,M., Christova,Y., Cervettini,D., Boge,F.L.,
            Liu,K.C., Buse,S., Maslen,S., Salmond,G.P.C. and Chin,J.W.
  TITLE     Sense Codon Reassignment Enables Viral Resistance and Encoded
            Polymer Synthesis
  JOURNAL   Science (2021) In press
REFERENCE   2  (bases 1 to 4347)
  AUTHORS   Robertson,W.E., Funke,L.F.H., de la Torre,D., Fredens,J.,
            Elliott,T.S., Spinck,M., Christova,Y., Cervettini,D., Boge,F.L.,
            Liu,K.C., Buse,S., Maslen,S., Salmond,G.P.C. and Chin,J.W.
  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
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4347
                     /organism="Cloning vector pMB1_MmPylRS-pylT-opt-serT.UGA"
                     /mol_type="other DNA"
                     /db_xref="taxon:2842476"
     regulatory      14..39
                     /regulatory_class="promoter"
                     /note="GlnRS promoter; GlnRS Promoter"
     CDS             81..1445
                     /note="codon-optimized; MmPylRS; from Methanosarcina
                     mazei"
                     /codon_start=1
                     /transl_table=11
                     /product="PylRS"
                     /protein_id="QWF36664.1"
                     /db_xref="GI:2049107078"
                     /translation="MDKKPLNTLISATGLWMSRTGTIHKIKHHEVSRSKIYIEMACGD
                     HLVVNNSRSSRTARALRHHKYRKTCKRCRVSDEDLNKFLTKANEDQTSVKVKVVSAPT
                     RTKKAMPKSVARAPKPLENTEAAQAQPSGSKFSPAIPVSTQESVSVPASVSTSISSIS
                     TGATASALVKGNTNPITSMSAPVQASAPALTKSQTDRLEVLLNPKDEISLNSGKPFRE
                     LESELLSRRKKDLQQIYAEERENYLGKLEREITRFFVDRGFLEIKSPILIPLEYIERM
                     GIDNDTELSKQIFRVDKNFCLRPMLAPNLYNYLRKLDRALPDPIKIFEIGPCYRKESD
                     GKEHLEEFTMLNFCQMGSGCTRENLESIITDFLNHLGIDFKIVGDSCMVYGDTLDVMH
                     GDLELSSAVVGPIPLDREWGIDKPWIGAGFGLERLLKVKHDFKNIKRAARSESYYNGI
                     STNL"
     regulatory      1980..2004
                     /regulatory_class="promoter"
                     /note="lpp promoter; Ipp p"
     tRNA            2025..2096
                     /product="tRNA-Pyl"
                     /note="Mb PylTwt (NC_007355.1); Mb PylTwt (NC_007355.1)"
     misc_feature    2047..2065
                     /note="ASL"
     regulatory      2103..2131
                     /regulatory_class="terminator"
                     /note="rrnC terminator; rrnC term"
     regulatory      2598..2645
                     /regulatory_class="terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase; T7 terminator"
     CDS             complement(2816..3631)
                     /note="confers resistance to kanamycin in bacteria or G418
                     (Geneticin(R)) in eukaryotes; KanR"
                     /codon_start=1
                     /transl_table=11
                     /product="aminoglycoside phosphotransferase"
                     /protein_id="QWF36665.1"
                     /db_xref="GI:2049107079"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
                     GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
                     GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
                     VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
                     RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     misc_feature    2906..2908
                     /note="TCA to AGT"
     misc_feature    3032..3034
                     /note="TCA to AGT"
     misc_feature    3056..3058
                     /note="TCA to AGT"
     misc_feature    3065..3067
                     /note="TCA to AGT"
     misc_feature    3170..3172
                     /note="TCA to AGT"
     misc_feature    3233..3235
                     /note="TCG to AGC"
     misc_feature    3284..3286
                     /note="TCA to AGT"
     misc_feature    3524..3526
                     /note="TCA to AGT"
     gene            complement(3632..3723)
                     /gene="bla"
     regulatory      complement(3632..3723)
                     /regulatory_class="promoter"
                     /gene="bla"
                     /note="AmpR promoter"
     rep_origin      complement(3787..4277)
                     /note="pMB1 origin of replication (clockwise) (RNAII -35
                     to RNA/DNA switch point); label: pMB1 ori"
                     /direction=LEFT
ORIGIN      
        1 ggatcctcgg gagttgtcag cctgtcccgc ttataagatc atacgccgtt atacgttgtt
       61 tacgctttga ggaatcccat atggacaaaa aaccgctgaa caccctgatc tctgctaccg
      121 gtctgtggat gtctcgtacc ggtaccatcc acaaaatcaa acaccacgaa gtttctcgtt
      181 ctaaaatcta catcgaaatg gcttgcggtg accacctggt tgttaacaac tctcgttctt
      241 ctcgtaccgc tcgtgctctg cgtcaccaca aataccgtaa aacctgcaaa cgttgccgtg
      301 tttctgacga agacctgaac aaattcctga ccaaagctaa cgaagaccag acctctgtta
      361 aagttaaagt tgtttctgct ccgacccgta ccaaaaaagc tatgccgaaa tctgttgctc
      421 gtgctccgaa accgctggaa aacaccgaag ctgctcaggc tcagccgtct ggttctaaat
      481 tctctccggc tatcccggtt tctacccagg aatctgtttc tgttccggct tctgtttcta
      541 cctctatctc ttctatctct accggtgcta ccgcttctgc tctggttaaa ggtaacacca
      601 acccgatcac ctctatgtct gctccggttc aggcttctgc tccggctctg accaaatctc
      661 agaccgaccg tctggaagtt ctgctgaacc cgaaagacga aatctctctg aactctggta
      721 aaccgttccg tgaactggaa tctgaactgc tgtctcgtcg taaaaaagac ttacaacaga
      781 tctacgctga agaacgtgaa aactacctgg gtaaactgga acgtgaaatc acgcgtttct
      841 tcgttgaccg tggtttcctg gaaatcaaat ctccgatcct gatcccgctg gaatacatcg
      901 aacgtatggg tatcgacaac gacaccgaac tgtctaaaca gatcttccgt gttgacaaaa
      961 acttctgcct gcgtccgatg ctggctccga acctgtacaa ctacctgcgt aaactggacc
     1021 gtgctctgcc ggacccgatc aaaatcttcg aaatcggtcc gtgctaccgt aaagaatctg
     1081 acggtaaaga acacctggaa gaattcacca tgctgaactt ctgccagatg ggttctggtt
     1141 gcacccgtga aaacctggaa tctatcatca ccgacttcct gaaccacctg ggtatcgact
     1201 tcaaaatcgt tggtgactct tgcatggttt acggtgacac cctggacgtt atgcacggtg
     1261 acctggaact gtcttctgct gttgttggtc cgatcccgct ggaccgtgaa tggggtatcg
     1321 acaaaccgtg gatcggtgct ggtttcggtc tggaacgtct gctgaaagtt aaacacgact
     1381 tcaaaaacat caaacgtgct gctcgttctg aatcttacta caacggtatc tctaccaacc
     1441 tgtaactgca gtttcaaacg ctaaattgcc tgatgcgcta cgcttatcag gcctacatga
     1501 tctctgcaat atattgagtt tgcgtgcttt tgtaggccgg ataaggcgtt cacgccgcat
     1561 ccggcaagaa acagcaaaca atccaaaacg ccgcgttcgg cggtcgacac agatgtaggt
     1621 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     1681 gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgctgcc
     1741 ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc
     1801 atgccccgcg cccaccggaa ggagctaccg gcagcggtgc ggactgttgt aactcagaat
     1861 aagaaatgag gccgctcatg gcgttctgtt gcccgtctca ctggtgaaaa gaaaaacaac
     1921 cctggcgccg cttctttgag cgaacgatca aaaataagtg gcgccccatc aaaaaaatat
     1981 tctcaacata aaaaactttg tgtaatactt gtaacgctag atctggaaac gtgatcatgt
     2041 agatcgaacg gtcttgaaaa ccgttcagtg gggttagatt ccccacgttt ccgccaacta
     2101 gtatccttag cgaaagctaa ggattttttt taagcttggc actggccgtc gttttacaac
     2161 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     2221 tcgccagacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     2281 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagcccgag atgcgccgcg
     2341 tgcggctgct ggagatggcg gacgcgatgg atatgttctg gcggccgcat aatgcttaag
     2401 tcgaacagaa agtaatcgta ttgtacacgg ccgcataatc gaaattaaat tggatatcgg
     2461 caggccacgc gatcgctgac gtcggtaccc tcgagtctgg taaagaaacc gctgctgcga
     2521 aatttgaacg ccagcacatg gactcgtcta ctagcgcagc ttaattaacc taggctgctg
     2581 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     2641 ttttgctgaa acctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata
     2701 aaccggtaaa ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga
     2761 ccgggtcatc gtggccggat cttgcggccc ctcggcttga acgaattgtt agacattaga
     2821 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     2881 atttttgaaa aagccgtttc tgtaaactag gagaaaactc accgaggcag ttccatagga
     2941 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     3001 atttcccctc gtcaaaaata aggttatcaa gactgaaatc accatgagtg acgacactat
     3061 ccggactgaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat
     3121 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcga ctttgcgcct
     3181 gagcgagacg aaatacgcgg tcgctgttaa aaggacaatt acaaacagga atgctatgca
     3241 accggcgcag gaacactgcc agcgcatcaa caatattttc accactatca ggatattctt
     3301 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
     3361 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     3421 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     3481 ctggcgcatc gggcttccca tacaatcgat agattgtcgc accactttgc ccgacattat
     3541 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag
     3601 agcaagacgt ttcccgttga atatggctca tactcttcct ttttcaatat tattgaagca
     3661 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     3721 aaatagctag cgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3781 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3841 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3901 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3961 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4021 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4081 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4141 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4201 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     4261 agttaccttc ggaaaaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4321 acgaaaactc acgttaaggg attttgg
//

