LOCUS       sequence-299372-        3058 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid with single copy of Mma PylT M15 mutant (UUA anticodon).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3058)
  AUTHORS   Meineke B, Heimgartner J, Eirich J, Landreh M, Elsasser SJ
  TITLE     Site-Specific Incorporation of Two ncAAs for Two-Color Bioorthogonal
            Labeling and Crosslinking of Proteins on Live Mammalian Cells.
  JOURNAL   Cell Rep. 2020 Jun 23;31(12):107811. doi: 
            10.1016/j.celrep.2020.107811.
  PUBMED    32579937
REFERENCE   2  (bases 1 to 3058)
  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: "Cell Rep.";
            date: "2020-06-23"; volume: "31(12):107811. doi"; pages: " 
            10.1016/j.celrep.2020.107811"
COMMENT     Sequence Label: pUC_h7SK M15 (UUA)
FEATURES             Location/Qualifiers
     source          1..3058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(884..900)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(884..900)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(897..919)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    908..924
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(932..938,939..956,957..962))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    977..998
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1115..1132)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1286..1874)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1366..1385)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2045..2860)
                     /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="MSHIQRETSCLKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2768..2787
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2861..2963)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3031..3049
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accaaatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcatcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgcaac gcgatgacga tggatagcga
      421 ttcatcgatg agctgacccg atcgccgccg ccggagggtt gcgtttgaga cgggcgacag
      481 attttctaga gcactagtgg ccttggagtg cagtatttag catgccccac ccatctgcaa
      541 ggcattctgg atagtgtcaa aacagccgga aatcaagtcc gtttatctca aactttagca
      601 ttttgggaat aaatgatatt tgctatgctg gttaaattag attttagtta aatttcctgc
      661 tgaagctcta gtacgataag taacttgacc taagtgtaaa gttgagattt ccttcaggtt
      721 tatatagctt gtgcgccgcc tgggtacctc ggaaacctgg tcagggagac cgaacggact
      781 ttaaatccgt tcagccgggt tcgattcccg gggtttccgg acaagtgcgg tttttatcag
      841 ttctggacca gcgagctgtg ctgcgactcg tggcgtaatc atggtcatag ctgtttcctg
      901 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
      961 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     1021 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     1081 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     1141 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     1201 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1261 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1321 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1381 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgtcgctt accggatacc
     1441 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1501 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1561 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1621 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1681 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     1741 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     1801 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1861 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     1921 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     1981 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     2041 acagttagaa aaactcatcg agcatcaagt gaaactgcaa tttattcata tcaggattat
     2101 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     2161 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     2221 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     2281 cgactgaatc cggtgagaat ggcaagagtt tatgcatttc tttccagact tgttcaacag
     2341 gccagccatt acgctcgtca tcaaaatcac tcgcaccaac caaaccgtta ttcattcgtg
     2401 attgcgcctg agcgagacga aatacgcgat cgccgttaaa aggacaatta caaacaggaa
     2461 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     2521 gatattcttc taatacctgg aatgctgttt tccctgggat cgcagtggtg agtaaccatg
     2581 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     2641 agtttagcct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     2701 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     2761 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     2821 gcggcttcaa gcaagacgtt tcccgttgaa tatggctcat tttagcttcc ttagctcctg
     2881 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     2941 tggaacctct tacgtgccga tcacacattt ccccgaaaag tgccacctga cgtctaagaa
     3001 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
