LOCUS       sequence-477156-        5652 bp    DNA     circular SYN 16-JUL-2025
DEFINITION  Site-specific incorporation of L-lactyl-lysine into target proteins 
            in E. coli cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5652)
  AUTHORS   Shao C, Tang S, Yu S, Liu C, Zhang Y, Wan T, He Z, Yuan Q, Wu S, 
            Zhang H, Wan N, Zhan M, Tan RX, Hao H, Ye H, Wang N
  TITLE     Genetic code expansion reveals site-specific lactylation in living 
            cells reshapes protein functions.
  JOURNAL   Nat Commun. 2025 Jan 8;16(1):227. doi: 10.1038/s41467-024-55165-2.
  PUBMED    39779673
REFERENCE   2  (bases 1 to 5652)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2025-01-8"; volume: "16(1):227. doi"; pages: " 
            10.1038/s41467-024-55165-2"
COMMENT     Sequence Label: pEvol-MmKlacRS1
FEATURES             Location/Qualifiers
     source          1..5652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..879)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPSDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        906..1190
                     /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     1113..1132
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2598..2615
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2671..2688)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(3674..4333)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4248..4267
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4334..4436)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4828..4845)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4962..5507)
                     /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(5042..5061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac ggaactcgct
       61 cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc
      121 aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg
      181 gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct ggcggaaaag
      241 atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga tatcaaaatt
      301 gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat tatccatcgg
      361 tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct caagcagatt
      421 tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga tttgcccaaa
      481 caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg tattggcaaa
      541 tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt aaacccactg
      601 gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc ctggcgggaa
      661 cagcaaaata tcactcggtc ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg
      721 accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt cgataaaaaa
      781 atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg cattaaacga
      841 gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac tcccgccatt
      901 cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg tcttttactg
      961 gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt aacaaagcgg
     1021 gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca
     1081 cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat ccataagatt
     1141 agcggatcct acctgacgct ttttatcgca actctctact gtttctccat acccgttttt
     1201 ttgggctaac aggaggaatt actagtatgg ataaaaagcc tttgaacact ctgatttctg
     1261 cgaccggtct gtggatgtcc cgcaccggca ccatccacaa aatcaaacac catgaagtta
     1321 gccgttccaa aatctacatt gaaatggctt gcggcgatca cctggttgtc aacaactccc
     1381 gttcttctcg taccgctcgc gcactgcgcc accacaaata tcgcaaaacc tgcaaacgtt
     1441 gccgtgttag cgatgaggac ctgaacaaat tcctgaccaa agctaacgag gatcagacct
     1501 ccgtaaaagt gaaggtagta agcgctccga cccgtactaa aaaggctatg ccaaaaagcg
     1561 tggcccgtgc cccgaaacct ctggaaaaca ccgaggcggc tcaggctcaa ccatccggtt
     1621 ctaaattttc tccggcgatc ccagtgtcca cccaagaatc tgtttccgta ccagcaagcg
     1681 tgtctaccag cattagcagc atttctaccg gtgctaccgc ttctgcgctg gtaaaaggta
     1741 acactaaccc gattactagc atgtctgcac cggtacaggc aagcgcccca gctctgacta
     1801 aatcccagac ggaccgtctg gaggtgctgc tgaacccaaa ggatgaaatc tctctgaaca
     1861 gcggcaagcc tttccgtgag ctggaaagcg agctgctgtc tcgtcgtaaa aaggatctgc
     1921 aacagatcta cgctgaggaa cgcgagaact atctgggtaa gctggagcgc gaaattactc
     1981 gcttcttcgt ggatcgcggt ttcctggaga tcaaatctcc gattctgatt ccgctggaat
     2041 acattgaacg tatgggcatc gataatgata ccgaactgtc taaacagatc ttccgtgtgg
     2101 ataaaaactt ctgtctgcgt ccgatgatgg ccccgaacct gctgaactat gcgcgtaaac
     2161 tggaccgtgc cctgccggac ccgatcaaaa ttttcgagat cggtccttgc taccgtaaag
     2221 agtccgacgg taaagagcac ctggaagaat tcaccatgct gaacttcttt cagatgggta
     2281 gcggttgcac gcgtgaaaac ctggaatcca ttatcaccga cttcctgaat cacctgggta
     2341 tcgatttcaa aattgttggt gacagctgta tggtgtatgg cgatacgctg gatgttatgc
     2401 acggcgatct ggagctgtct tccgcagtag tgggcccaat cccgctggat cgtgagtggg
     2461 gtatcgacaa accttggatc ggtgcgggtt ttggtctgga gcgtctgctg aaagtaaaac
     2521 acgacttcaa gaacatcaaa cgtgctgcac gttccgagtc ctattacaat ggtatttcta
     2581 ctaacctgta agtcgaccat catcatcatc atcattgagt ttaaacggtc tccagcttgg
     2641 ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag
     2701 cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat
     2761 gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag
     2821 agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggccttgt
     2881 ttgtgagctc ccggtcatca atcatcccca taatccttgt tagattatca attttaaaaa
     2941 actaacagtt gtcagcctgt cccgctttaa tatcatacgc cgttatacgt tgtttacgct
     3001 ttgaggaatc ccatatgata cgcggccgct ttcaaacgct aaattgcctg atgcgctacg
     3061 cttatcaggc ctacatgatc tctgcaatat attgagtttg cgtgcttttg taggccggat
     3121 aaggcgttca cgccgcatcc ggcaagaaac agcaaacaat ccaaaacgcc gcgttcagcg
     3181 gcgttttttc tgcttttctt cgcgaattaa ttccgcttcg caacatgtga gcaccggttt
     3241 attgactacc ggaagcagtg tgaccgtgtg cttctcaaat gcctgaggcc agtttgctca
     3301 ggctctcccc gtggaggtaa taattgacga tatgatcagt gcacggctaa ctaagcggcc
     3361 tgctgacttt ctcgccgatc aaaaggcatt ttgctattaa gggattgacg agggcgtatc
     3421 tgcgcagtaa gatgcgcccc gcattggaaa cctgatcatg tagatcgaat ggactctaaa
     3481 tccgttcagc cgggttagat tcccggggtt tccgccaaat tcgaaaagcc tgctcaacga
     3541 gcaggctttt ttgcatgctc gagcagctca gggtcgaatt tgctttcgaa tttctgccat
     3601 tcatccgctt attatcactt attcaggcgt agcaaccagg cgtttaaggg caccaataac
     3661 tgccttaaaa aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa
     3721 gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca
     3781 tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt
     3841 tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga
     3901 cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg
     3961 ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga
     4021 gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc
     4081 atatcaccag ctcaccgtct ttcattgcca tacggaattc cggatgagca ttcatcaggc
     4141 gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta
     4201 aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa
     4261 atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga
     4321 tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc
     4381 ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt
     4441 ctcattttcg ccaaaagttg gcccagggct tcccggtatc aacagggaca ccaggattta
     4501 tttattctgc gaagtgatct tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg
     4561 atgctgccaa cttactgatt tagtgtatga tggtgttttt gaggtgctcc agtggcttct
     4621 gtttctatca gctgtccctc ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc
     4681 accgccggac atcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg
     4741 gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa
     4801 tatgtgatac aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg
     4861 actgcggcga gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa
     4921 gatacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc
     4981 cctgacaagc atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta
     5041 taaagatacc aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt
     5101 cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact
     5161 cagttccggg taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc
     5221 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa
     5281 agcaccactg gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg
     5341 ccggttaagg ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac
     5401 ctcggttcaa agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt
     5461 tttcgttttc agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta
     5521 ttaatcagat aaaatatttc tagatttcag tgcaatttat ctcttcaaat gtagcacctg
     5581 aagtcagccc catacgatat aagttgtaat tctcatgttt gacagcttat catcgataag
     5641 cttggtaccc aa
//
