LOCUS       sequence-443626-        2797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Methanomethylophilus alvus pyrrolysine tRNA synthetase 
            engineered for 3-nitroY and 3-haloY under GlnS promoter. Used as a 
            control for Ma Pyl tRNA-synthetase selections..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2797)
  TITLE     Engineering of Aminoacyl-tRNA synthetase/tRNA pairs for efficient 
            genetic encoding of non-canonical amino acids into proteins.
  JOURNAL   Unpublished
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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pBK-Ma pylRS nitroY/haloY-F5
FEATURES             Location/Qualifiers
     source          1..2797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     374..393
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      1385..1394
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1745..2560)
                     /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             complement(1745..2560)
                     /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     2468..2487
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2552..2571
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(2682..2797,1..473)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
       61 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
      121 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
      181 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
      241 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
      301 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      361 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      421 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      481 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtgcgaag
      541 cggaattaat tcgcgaagaa aagcagaaaa aacgccgctg aacgcggcgt tttggattgt
      601 ttgctgtttc ttgccggatg cggcgtgaac gccttatccg gcctacaaaa gcacgcaaac
      661 tcaatatatt gcagagatca tgtaggcctg ataagcgtag cgcatcaggc aatttagcgt
      721 ttgaaactgc aggcttcagc cgctgttgat cttggctcca ttcaagtagg agatggaggc
      781 gccgcctttc ttcacagtgc tgtacttttc tcttatggtc agcagcctct ccaggccaaa
      841 gcccgctcca ctacggggtt catgcacgtc gtgggcggcg tccaggtagt gggggcccac
      901 acaagcagag cacacctcct ggccattgat ctccacatca atggtttctt tgtagacatc
      961 gctctcttcc tgcaccaaat catagtcggg cagtccagca gccttcatga ccactgagat
     1021 gtagttcttc agcacttctg tggcatctcc tcttggcccc atgtccgaca gagccagcat
     1081 ggtgaactcc tccaggtgca ttccgctgtg gctttctttc ctaaagcagg agcccatttc
     1141 gaaaattttc acagggccgt ccgtgtggtc gcgcaggtcc cgcatcacgg agtaaaggtt
     1201 gggggcgagc atgggcctca gagccctctt ctcatcgatc cagaacacct gcttgaacag
     1261 gggcttgtcc tcggtgatgg tcatgcgagc cagcgcgtcc ttagaaatga agatgggggt
     1321 cctgacctcg atgaagccct cggccaccag ggcgtcggcg atgtcattca tcagctgggt
     1381 cagcccatgg cggctggggt tggcaatcat gcccttgatc tttttctcgt tgtctgtgga
     1441 agccacggac atctccttgg agaaggctgc atctctagag gccagatctt caaacacctt
     1501 ctgctcatag gtgccgttgc catactcccg caggcgctgg atctgggcat ctgtgtattt
     1561 cactgtcata tgggattcct caaagcgtaa acaacgtata acggcgtatg atcttataag
     1621 cgggacaggc tgacaacccg aggatccttc aactcagcaa aagttcgatt tattcaacaa
     1681 agccgccgtc ccgtcaagtc agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt
     1741 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     1801 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     1861 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     1921 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     1981 cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact tgttcaacag
     2041 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     2101 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     2161 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     2221 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     2281 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     2341 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     2401 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     2461 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     2521 gcggcctcga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     2581 ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc
     2641 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2701 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2761 gatcaagagc taccaactct ttttccgaag gtaactg
//
