LOCUS       sequence-359663-        5652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N‐acetyl lysyl‐tRNA synthetase with cognate tRNA for genetic code 
            expansion with ochre suppression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5652)
  AUTHORS   Wu B, Wang Z, Huang Y, Liu WR
  TITLE     Catalyst-free and site-specific one-pot dual-labeling of a protein 
            directed by two genetically incorporated noncanonical amino acids.
  JOURNAL   Chembiochem. 2012 Jul 9;13(10):1405-8. doi: 10.1002/cbic.201200281. 
            Epub 2012 May 24.
  PUBMED    22628069
REFERENCE   2  (bases 1 to 5652)
  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: 
            "10.1002/cbic.201200281"; journalName: "Chembiochem"; date: 
            "2012-07-9- 9"; volume: "13"; issue: "10"; pages: "1405-8"
COMMENT     Sequence Label: pEVOL-MmAcKRS1-PylTUUA
FEATURES             Location/Qualifiers
     source          1..5652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(480..497)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(614..1159)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(694..713)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1305..2183)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPSDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2210..2494
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     2417..2436
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3902..3919
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3975..3992)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(4978..5637)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5552..5571
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(5638..5652,1..88))
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
       61 aaagttggaa cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg
      121 cttcccggta tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac
      181 aggtatttat tcggcgcaaa gtgcgtcggg tgatgctgcc aacttactga tttagtgtat
      241 gatggtgttt ttgaggtgct ccagtggctt ctgtttctat cagctgtccc tcctgttcag
      301 ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg acatcagcgc tagcggagtg
      361 tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc atgtggcagg
      421 agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata ttccgcttcc
      481 tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa
      541 cggggcggag atttcctgga agatgccagg aagatactta acagggaagt gagagggccg
      601 cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct
      661 caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg
      721 gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat
      781 ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag
      841 ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat
      901 cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat
      961 tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag
     1021 ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag
     1081 aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg
     1141 cagaccaaaa cgatctcaag aagatcatct tattaatcag ataaaatatt tctagatttc
     1201 agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgta
     1261 attctcatgt ttgacagctt atcatcgata agcttggtac ccaattatga caacttgacg
     1321 gctacatcat tcactttttc ttcacaaccg gcacggaact cgctcgggct ggccccggtg
     1381 cattttttaa atacccgcga gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg
     1441 gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg cctggctgat acgttggtcc
     1501 tcgcgccagc ttaagacgct aatccctaac tgctggcgga aaagatgtga cagacgcgac
     1561 ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga
     1621 tcgctgatgt actgacaagc ctcgcgtacc cgattatcca tcggtggatg gagcgactcg
     1681 ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca gatttatcgc cagcagctcc
     1741 gaatagcgcc cttccccttg cccggcgtta atgatttgcc caaacaggtc gctgaaatgc
     1801 ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg caaatattga cggccagtta
     1861 agccattcat gccagtaggc gcgcggacga aagtaaaccc actggtgata ccattcgcga
     1921 gcctccggat gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa aatatcactc
     1981 ggtcggcaaa caaattctcg tccctgattt ttcaccaccc cctgaccgcg aatggtgaga
     2041 ttgagaatat aacctttcat tcccagcggt cggtcgataa aaaaatcgag ataaccgttg
     2101 gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa acgagtatcc cggcagcagg
     2161 ggatcatttt gcgcttcagc catacttttc atactcccgc cattcagaga agaaaccaat
     2221 tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt actggctctt ctcgctaacc
     2281 aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca
     2341 aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag tccacattga ttatttgcac
     2401 ggcgtcacac tttgctatgc catagcattt ttatccataa gattagcgga tcctacctga
     2461 cgctttttat cgcaactctc tactgtttct ccatacccgt ttttttgggc taacaggagg
     2521 aattactagt atggataaga aaccgctgaa tactctgatt tctgcaactg gtctgtggat
     2581 gagccatacc ggcaccatcc acaagatcaa acaccgcgag gtttcccgaa gcaaaatcta
     2641 catcgaaatg gcgtgcggtg accacctggt ggtaaacaac tcccgttctt ctcgtactgc
     2701 acgtgctctg cgccaccaca agtaccgtaa gacctgcaag cgctgtcgcg tgtctgatga
     2761 agacctgaac aaattcctga ctaaagcgaa cgaagatcag acttctgtga aggtgaaagt
     2821 tgtttctgcc ccaacccgca ccaagaaagc gatgccgaag tccgttgcac gcgctccgaa
     2881 accgctggag aactccgaag ccgcacaggc ccagccgtct ggttctaagt tttctccggc
     2941 aatcccggtt tctactcagg agtctgtgtc tgtgccagct tctgttagca cttctatttc
     3001 ctctatcagc actggtgcga ctgcgtccgc tctggtaaaa ggtaacacta acccgatcac
     3061 cagcatgtct gctccggttc aggcttctgc accggcactg actaaaagcc agactgaccg
     3121 tctggaggtt ctgctgaacc cgaaagatga aatcagcctg aactctggca aaccgttccg
     3181 tgaactggaa tccgaactgc tgtctcgtcg taagaaagac ctgcaacaaa tctatgctga
     3241 agagcgtgaa aactacctgg gtaaactgga acgtgaaatc acccgtttct ttgtggaccg
     3301 tggtttcctg gaaatcaagt ctccgatcct gatcccgctg gaatacatcg agcgcatggg
     3361 tattgataac gacaccgaac tgtccaagca gattttccgt gtggacaaga acttctgcct
     3421 gcgtccgatg atggcaccga acctgctgaa ttacgcacgt aaactggatc gtgcactgcc
     3481 ggacccgatc aaaatctttg aaatcggtcc atgctatcgt aaggagagcg acggtaaaga
     3541 acacctggaa gagttcacta tgctgaactt ttttcagatg ggttctggct gcacccgtga
     3601 aaatctggaa tctatcatca ccgacttcct gaaccacctg ggcattgact tcaaaatcgt
     3661 tggtgattcc tgcatggttt acggtgacac tctggacgtt atgcatggtg atctggaact
     3721 gagcagcgct gttgtgggtc cgattccgct ggatcgtgaa tggggtatcg ataaaccgtg
     3781 gattggtgct ggcttcggtc tggaacgtct gctgaaagtt aagcacgact ttaagaacat
     3841 caaacgtgct gcgcgttccg agtcctatta caacggcatt agcactaacc tgtaagtcga
     3901 ccatcatcat catcatcatt gagtttaaac ggtctccagc ttggctgttt tggcggatga
     3961 gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag
     4021 aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgtcgaa ctcagaagtg
     4081 aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag
     4141 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc ttgtttgtga gctcccggtc
     4201 atcaatcatc cccataatcc ttgttagatt atcaatttta aaaaactaac agttgtcagc
     4261 ctgtcccgct ttaatatcat acgccgttat acgttgttta cgctttgagg aatcccatat
     4321 gatacgcggc cgctttcaaa cgctaaattg cctgatgcgc tacgcttatc aggcctacat
     4381 gatctctgca atatattgag tttgcgtgct tttgtaggcc ggataaggcg ttcacgccgc
     4441 atccggcaag aaacagcaaa caatccaaaa cgccgcgttc agcgacgttt tttctgcttt
     4501 tcttcgcgaa ttaattccgc ttcgcaacat gtgagcaccg gtttattgac taccggaagc
     4561 agtgtgaccg tgtgcttctc aaatgcctga ggccagtttg ctcaggctct ccccgtggag
     4621 gtaataattg acgatatgat cagtgcacgg ctaactaagc ggcctgctga ctttctcgcc
     4681 gatcaaaagg cattttgcta ttaagggatt gacgagggcg tatctgcgca gtaagatgcg
     4741 ccccgcattg gaaacctgat catgtagatc gaatggactt taaatccgtt cagccgggtt
     4801 agattcccgg ggtttccgcc aaattcgaaa agcctgctca acgagcaggc ttttttgcat
     4861 gctcgagcag ctcagggtcg aatttgcttt cgaatttctg ccattcatcc gcttattatc
     4921 acttattcag gcgtagcaac caggcgttta agggcaccaa taactgcctt aaaaaaatta
     4981 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
     5041 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
     5101 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac
     5161 gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc
     5221 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata
     5281 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc
     5341 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc
     5401 gtctttcatt gccatacgga attccggatg agcattcatc aggcgggcaa gaatgtgaat
     5461 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc
     5521 cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc
     5581 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt
     5641 agcttcctta gc
//
