LOCUS       sequence-272335-        5652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sequence optimized N‐acetyl lysyl‐tRNA synthetase with cognate tRNA 
            for genetic code expansion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5652)
  AUTHORS   Wang WW, Angulo-Ibanez M, Lyu J, Kurra Y, Tong Z, Wu B, Zhang L, 
            Sharma V, Zhou J, Lin H, Gao YQ, Li W, Chua KF, Liu WR
  TITLE     A Click Chemistry Approach Reveals the Chromatin-Dependent Histone 
            H3K36 Deacylase Nature of SIRT7.
  JOURNAL   J Am Chem Soc. 2019 Feb 13;141(6):2462-2473. doi: 
            10.1021/jacs.8b12083. Epub 2019 Feb 4.
  PUBMED    30653310
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: 
            "10.1021/jacs.8b12083"; journalName: "J Am Chem Soc"; date: 
            "2019-02-13- 13"; volume: "141"; issue: "6"; pages: "2462-2473"
COMMENT     Sequence Label: pEVOL-AckRS
FEATURES             Location/Qualifiers
     source          1..5652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /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     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1693..1710
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1766..1783)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(2769..3428)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3343..3362
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3429..3531)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3923..3940)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4057..4602)
                     /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(4137..4156)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4748..5626)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPSDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
      301 ctaacaggag gaattactag tatggataag aaaccgctga atactctgat ttctgcaact
      361 ggtctgtgga tgagccgtac cggcaccatc cacaagatca aacaccacga ggtttcccgt
      421 agcaaaatct acatcgaaat ggcgtgcggt gaccacctgg tggtaaacaa ctcccgttct
      481 tctcgtactg cacgtgctct gcgccaccac aagtaccgta agacctgcaa gcgctgtcgc
      541 gtgtctgatg aagacctgaa caaattcctg actaaagcga acgaagatca gacttctgtg
      601 aaggtgaaag ttgtttctgc cccaacccgc accaagaaag cgatgccgaa gtccgttgca
      661 cgcgctccga aaccgctgga gaacaccgaa gccgcacagg cccagccgtc tggttctaag
      721 ttttctccgg caatcccggt ttctactcag gagtctgtgt ctgtgccagc ttctgttagc
      781 acttctattt cctctatcag cactggtgcg actgcgtccg ctctggtaaa aggtaacact
      841 aacccgatca ccagcatgtc tgctccggtt caggcttctg caccggcact gactaaaagc
      901 cagactgacc gtctggaggt tctgctgaac ccgaaagatg aaatcagcct gaactctggc
      961 aaaccgttcc gtgaactgga atccgaactg ctgtctcgtc gtaagaaaga cctgcaacaa
     1021 atctatgctg aagagcgtga aaactacctg ggtaaactgg aacgtgaaat cacccgtttc
     1081 tttgtggacc gtggtttcct ggaaatcaag tctccgatcc tgatcccgct ggaatacatc
     1141 gagcgcatgg gtattgataa cgacaccgaa ctgtccaagc agattttccg tgtggacaag
     1201 aacttctgcc tgcgtccgat gatggcaccg aacctgctga attacgcgcg taaactggat
     1261 cgtgcactgc cggacccgat caaaatcttt gaaatcggtc catgctatcg taaggagagc
     1321 gacggtaaag aacacctgga agagttcact atgctgaact tttttcagat gggttctggc
     1381 tgcacccgtg aaaatctgga atctatcatc accgacttcc tgaaccacct gggcattgac
     1441 ttcaaaatcg ttggtgattc ctgcatggtt tacggtgaca ctctggacgt tatgcatggt
     1501 gatctggaac tgagcagcgc tgttgtgggt ccgattccgc tggatcgtga atggggtatc
     1561 gataaaccgt ggattggtgc tggcttcggt ctggaacgtc tgctgaaagt taagcacgac
     1621 tttaagaaca tcaaacgtgc tgcgcgttcc gagtcctatt acaacggcat tagcactaac
     1681 ctgtaagtcg accatcatca tcatcatcat tgagtttaaa cggtctccag cttggctgtt
     1741 ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc
     1801 tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga
     1861 actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag
     1921 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc cttgtttgtg
     1981 agctcccggt catcaatcat ccccataatc cttgttagat tatcaatttt aaaaaactaa
     2041 cagttgtcag cctgtcccgc tttaatatca tacgccgtta tacgttgttt acgctttgag
     2101 gaatcccata tgatacgcgg ccgctttcaa acgctaaatt gcctgatgcg ctacgcttat
     2161 caggcctaca tgatctctgc aatatattga gtttgcgtgc ttttgtaggc cggataaggc
     2221 gttcacgccg catccggcaa gaaacagcaa acaatccaaa acgccgcgtt cagcggcgtt
     2281 ttttctgctt ttcttcgcga attaattccg cttcgcaaca tgtgagcacc ggtttattga
     2341 ctaccggaag cagtgtgacc gtgtgcttct caaatgcctg aggccagttt gctcaggctc
     2401 tccccgtgga ggtaataatt gacgatatga tcagtgcacg gctaactaag cggcctgctg
     2461 actttctcgc cgatcaaaag gcattttgct attaagggat tgacgagggc gtatctgcgc
     2521 agtaagatgc gccccgcatt ggaaacctga tcatgtagat cgaatggact ctaaatccgt
     2581 tcagccgggt tagattcccg gggtttccgc caaattcgaa aagcctgctc aacgagcagg
     2641 cttttttgca tgctcgagca gctcagggtc gaatttgctt tcgaatttct gccattcatc
     2701 cgcttattat cacttattca ggcgtagcaa ccaggcgttt aagggcacca ataactgcct
     2761 taaaaaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt
     2821 ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc
     2881 accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc
     2941 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa
     3001 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca
     3061 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
     3121 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
     3181 accagctcac cgtctttcat tgccatacgg aattccggat gagcattcat caggcgggca
     3241 agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag
     3301 gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc
     3361 tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt
     3421 ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt
     3481 agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc aacgtctcat
     3541 tttcgccaaa agttggccca gggcttcccg gtatcaacag ggacaccagg atttatttat
     3601 tctgcgaagt gatcttccgt cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct
     3661 gccaacttac tgatttagtg tatgatggtg tttttgaggt gctccagtgg cttctgtttc
     3721 tatcagctgt ccctcctgtt cagctactga cggggtggtg cgtaacggca aaagcaccgc
     3781 cggacatcag cgctagcgga gtgtatactg gcttactatg ttggcactga tgagggtgtc
     3841 agtgaagtgc ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt
     3901 gatacaggat atattccgct tcctcgctca ctgactcgct acgctcggtc gttcgactgc
     3961 ggcgagcgga aatggcttac gaacggggcg gagatttcct ggaagatgcc aggaagatac
     4021 ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc cataggctcc gcccccctga
     4081 caagcatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag gactataaag
     4141 ataccaggcg tttccccctg gcggctccct cgtgcgctct cctgttcctg cctttcggtt
     4201 taccggtgtc attccgctgt tatggccgcg tttgtctcat tccacgcctg acactcagtt
     4261 ccgggtaggc agttcgctcc aagctggact gtatgcacga accccccgtt cagtccgacc
     4321 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac
     4381 cactggcagc agccactggt aattgattta gaggagttag tcttgaagtc atgcgccggt
     4441 taaggctaaa ctgaaaggac aagttttggt gactgcgctc ctccaagcca gttacctcgg
     4501 ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg
     4561 ttttcagagc aagagattac gcgcagacca aaacgatctc aagaagatca tcttattaat
     4621 cagataaaat atttctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc
     4681 agccccatac gatataagtt gtaattctca tgtttgacag cttatcatcg ataagcttgg
     4741 tacccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa ccggcacgga
     4801 actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag agttgatcgt
     4861 caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc aaaagcagct
     4921 tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct aactgctggc
     4981 ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg ctggcgatat
     5041 caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt acccgattat
     5101 ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac aattgctcaa
     5161 gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg ttaatgattt
     5221 gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag aaccccgtat
     5281 tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga cgaaagtaaa
     5341 cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga atctctcctg
     5401 gcgggaacag caaaatatca ctcggtcggc aaacaaattc tcgtccctga tttttcacca
     5461 ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc ggtcggtcga
     5521 taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc agatgggcat
     5581 taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt ttcatactcc
     5641 cgccattcag ag
//
