LOCUS       sequence-299370-        3059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid with single copy of Mma PylT (UUA anticodon).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3059)
  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 3059)
  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 Mma PylT (UUA)
FEATURES             Location/Qualifiers
     source          1..3059
                     /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(885..901)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(885..901)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(898..920)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    909..925
                     /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(933..939,940..957,958..963))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    978..999
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1116..1133)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1287..1875)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1367..1386)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2046..2861)
                     /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     2769..2788
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2862..2964)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3032..3050
                     /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 ccttggagct gcagtattta gcatgcccca cccatctgca
      541 aggcattctg gatagtgtca aaacagccgg aaatcaagtc cgtttatctc aaactttagc
      601 attttgggaa taaatgatat ttgctatgct ggttaaatta gattttagtt aaatttcctg
      661 ctgaagctct agtacgataa gtaacttgac ctaagtgtaa agttgagatt tccttcaggt
      721 ttatatagct tgtgcgccgc ctgggtacct cggaaacctg atcatgtaga tcgaacggac
      781 tttaaatccg ttcagccggg ttagattccc ggggtttccg gacaagtgcg gtttttatca
      841 gttctggacc agcgagctgt gctgcgactc gtggcgtaat catggtcata gctgtttcct
      901 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
      961 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     1021 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     1081 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     1141 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     1201 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     1261 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1321 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1381 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgtcgct taccggatac
     1441 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1501 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1561 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1621 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1681 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     1741 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     1801 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     1861 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     1921 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     1981 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2041 gacagttaga aaaactcatc gagcatcaag tgaaactgca atttattcat atcaggatta
     2101 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     2161 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     2221 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     2281 acgactgaat ccggtgagaa tggcaagagt ttatgcattt ctttccagac ttgttcaaca
     2341 ggccagccat tacgctcgtc atcaaaatca ctcgcaccaa ccaaaccgtt attcattcgt
     2401 gattgcgcct gagcgagacg aaatacgcga tcgccgttaa aaggacaatt acaaacagga
     2461 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     2521 ggatattctt ctaatacctg gaatgctgtt ttccctggga tcgcagtggt gagtaaccat
     2581 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     2641 cagtttagcc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     2701 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     2761 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     2821 cgcggcttca agcaagacgt ttcccgttga atatggctca ttttagcttc cttagctcct
     2881 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag
     2941 ttggaacctc ttacgtgccg atcacacatt tccccgaaaa gtgccacctg acgtctaaga
     3001 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc
//
