LOCUS       sequence-388117-        2797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses wild-type Methanomethylophilus alvus pyrrolysine tRNA 
            synthetase under GlnS promoter. Used for Pyl tRNA-synthetase 
            selections..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2797)
  AUTHORS   Avila-Crump S, Hemshorn ML, Jones CM, Mbengi L, Meyer K, Griffis JA,
            Jana S, Petrina GE, Pagar VV, Karplus PA, Petersson EJ, Perona JJ, 
            Mehl RA, Cooley RB
  TITLE     Generating Efficient Methanomethylophilus alvus Pyrrolysyl-tRNA 
            Synthetases for Structurally Diverse Non-Canonical Amino Acids.
  JOURNAL   ACS Chem Biol. 2022 Dec 16;17(12):3458-3469. doi: 
            10.1021/acschembio.2c00639. Epub 2022 Nov 16.
  PUBMED    36383641
REFERENCE   2  (bases 1 to 2797)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acschembio.2c00639"; journalName: "ACS Chem Biol"; date: 
            "2022-12-16- 16"; volume: "17"; issue: "12"; pages: "3458-3469"
COMMENT     Sequence Label: pBK- Ma PylRS WT
FEATURES             Location/Qualifiers
     source          1..2797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      249..258
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      complement(1170..1758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1250..1269)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1869..1888)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1880..2695
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             1880..2695
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1953..1972)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ctcgggttgt cagcctgtcc cgcttataag atcatacgcc gttatacgtt gtttacgctt
       61 tgaggaatcc catatgacag tgaaatacac agatgcccag atccagcgcc tgcgggagta
      121 tggcaacggc acctatgagc agaaggtgtt tgaagatctg gcctctagag atgcagcctt
      181 ctccaaggag atgtccgtgg cttccacaga caacgagaaa aagatcaagg gcatgattgc
      241 caaccccagc cgccatgggc tgacccagct gatgaatgac atcgccgacg ccctggtggc
      301 cgagggcttc atcgaggtca ggacccccat cttcatttct aaggacgcgc tggctcgcat
      361 gaccatcacc gaggacaagc ccctgttcaa gcaggtgttc tggatcgatg agaagagggc
      421 tctgaggccc atgctcgccc ccaacctgta ctccgtgatg cgggacctgc gcgaccacac
      481 ggacggccct gtgaaaattt tcgaaatggg ctcctgcttt aggaaagaaa gccacagcgg
      541 aatgcacctg gaggagttca ccatgctgaa tctggtggac atggggccaa gaggagatgc
      601 cacagaagtg ctgaagaact acatctcagt ggtcatgaag gctgctggac tgcccgacta
      661 tgatttggtg caggaagaga gcgatgtcta caaagaaacc attgatgtgg agatcaatgg
      721 ccaggaggtg tgctctgctg ccgtgggccc ccactacctg gacgccgccc acgacgtgca
      781 tgaaccctgg agtggagcgg gctttggcct ggagaggctg ctgaccataa gagaaaagta
      841 cagcactgtg aagaaaggcg gcgcctccat ctcctacttg aatggagcca agatcaacag
      901 cggctgaagc ctgcagtttc aaacgctaaa ttgcctgatg cgctacgctt atcaggccta
      961 catgatctct gcaatatatt gagtttgcgt gcttttgtag gccggataag gcgttcacgc
     1021 cgcatccggc aagaaacagc aaacaatcca aaacgccgcg ttcagcggcg ttttttctgc
     1081 ttttcttcgc gaattaattc cgcttcgcac atgtgagcaa aaggccagca aaaggccagg
     1141 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1201 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1261 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1321 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1381 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1441 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1501 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1561 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     1621 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     1681 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     1741 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     1801 aacgaaaact cacgttaagg gattttggtc atgaacaata aaactgtctg cttacataaa
     1861 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct cgaggccgcg
     1921 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     1981 gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct
     2041 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     2101 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     2161 atggttactc accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc
     2221 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     2281 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     2341 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     2401 tgttgaacaa gtctggaaag aaatgcataa gcttttgcca ttctcaccgg attcagtcgt
     2461 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     2521 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     2581 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     2641 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     2701 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcggctt
     2761 tgttgaataa atcgaacttt tgctgagttg aaggatc
//
