LOCUS       Exported                5652 bp ds-DNA     circular SYN 28-JUL-2021
DEFINITION  For E.Coli expression, 306L309L348I384F411W.
ACCESSION   .
VERSION     .
KEYWORDS    pEVOL-pylT-AznLRS
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5652)
  AUTHORS   Wang ZA, Kurra Y, Wang X, Zeng Y, Lee YJ, Sharma V, Lin H, Dai SY, 
            Liu WR
  TITLE     A Versatile Approach for Site-Specific Lysine Acylation in Proteins.
  JOURNAL   Angew Chem Int Ed Engl. 2017 Feb 1;56(6):1643-1647. doi: 
            10.1002/anie.201611415. Epub 2017 Jan 2.
  PUBMED    28042700
REFERENCE   2  (bases 1 to 5652)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 28, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5652
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(597..1475)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPSDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1502..1786
                     /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     1709..1728
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3194..3211
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3267..3284)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(4270..4929)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4844..4863
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4930..5032)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(5424..5441)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5558..451)
                     /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(5638..5)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca
       61 ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca
      121 gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta
      181 tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca
      241 gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac
      301 tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt
      361 ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca
      421 agagattacg cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata
      481 tttctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg
      541 atataagttg taattctcat gtttgacagc ttatcatcga taagcttggt acccaattat
      601 gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa ctcgctcggg
      661 ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc aaaaccaaca
      721 ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt cgcctggctg
      781 atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg gaaaagatgt
      841 gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg
      901 tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc catcggtgga
      961 tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag cagatttatc
     1021 gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg cccaaacagg
     1081 tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt
     1141 gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac ccactggtga
     1201 taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg cgggaacagc
     1261 aaaatatcac tcggtcggca aacaaattct cgtccctgat ttttcaccac cccctgaccg
     1321 cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat aaaaaaatcg
     1381 agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt aaacgagtat
     1441 cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc gccattcaga
     1501 gaagaaacca attgtccata ttgcatcaga cattgccgtc actgcgtctt ttactggctc
     1561 ttctcgctaa ccaaaccggt aaccccgctt attaaaagca ttctgtaaca aagcgggacc
     1621 aaagccatga caaaaacgcg taacaaaagt gtctataatc acggcagaaa agtccacatt
     1681 gattatttgc acggcgtcac actttgctat gccatagcat ttttatccat aagattagcg
     1741 gatcctacct gacgcttttt atcgcaactc tctactgttt ctccataccc gtttttttgg
     1801 gctaacagga ggaattacta gtatggataa gaaaccgctg aatactctga tttctgcaac
     1861 tggtctgtgg atgagccgta ccggcaccat ccacaagatc aaacaccacg aggtttcccg
     1921 tagcaaaatc tacatcgaaa tggcgtgcgg tgaccacctg gtggtaaaca actcccgttc
     1981 ttctcgtact gcacgtgctc tgcgccacca caagtaccgt aagacctgca agcgctgtcg
     2041 cgtgtctgat gaagacctga acaaattcct gactaaagcg aacgaagatc agacttctgt
     2101 gaaggtgaaa gttgtttctg ccccaacccg caccaagaaa gcgatgccga agtccgttgc
     2161 acgcgctccg aaaccgctgg agaacaccga agccgcacag gcccagccgt ctggttctaa
     2221 gttttctccg gcaatcccgg tttctactca ggagtctgtg tctgtgccag cttctgttag
     2281 cacttctatt tcctctatca gcactggtgc gactgcgtcc gctctggtaa aaggtaacac
     2341 taacccgatc accagcatgt ctgctccggt tcaggcttct gcaccggcac tgactaaaag
     2401 ccagactgac cgtctggagg ttctgctgaa cccgaaagat gaaatcagcc tgaactctgg
     2461 caaaccgttc cgtgaactgg aatccgaact gctgtctcgt cgtaagaaag acctgcaaca
     2521 aatctatgct gaagagcgtg aaaactacct gggtaaactg gaacgtgaaa tcacccgttt
     2581 ctttgtggac cgtggtttcc tggaaatcaa gtctccgatc ctgatcccgc tggaatacat
     2641 cgagcgcatg ggtattgata acgacaccga actgtccaag cagattttcc gtgtggacaa
     2701 gaacttctgc ctgcgtccga tgctggcacc gaacctgctg aattacctgc gtaaactgga
     2761 tcgtgcactg ccggacccga tcaaaatctt tgaaatcggt ccatgctatc gtaaggagag
     2821 cgacggtaaa gaacacctgg aagagttcac tatgctgaac tttattcaga tgggttctgg
     2881 ctgcacccgt gaaaatctgg aatctatcat caccgacttc ctgaaccacc tgggcattga
     2941 cttcaaaatc gttggtgatt cctgcatggt ttttggtgac actctggacg ttatgcatgg
     3001 tgatctggaa ctgagcagcg ctgttgtggg tccgattccg ctggatcgtg aatggggtat
     3061 cgataaaccg tggattggtg ctggcttcgg tctggaacgt ctgctgaaag ttaagcacga
     3121 ctttaagaac atcaaacgtg ctgcgcgttc cgagtcctat tacaacggca ttagcactaa
     3181 cctgtaagtc gaccatcatc atcatcatca ttgagtttaa acggtctcca gcttggctgt
     3241 tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     3301 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     3361 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     3421 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg ccttgtttgt
     3481 gagctcccgg tcatcaatca tccccataat ccttgttaga ttatcaattt taaaaaacta
     3541 acagttgtca gcctgtcccg ctttaatatc atacgccgtt atacgttgtt tacgctttga
     3601 ggaatcccat atgatacgcg gccgctttca aacgctaaat tgcctgatgc gctacgctta
     3661 tcaggcctac atgatctctg caatatattg agtttgcgtg cttttgtagg ccggataagg
     3721 cgttcacgcc gcatccggca agaaacagca aacaatccaa aacgccgcgt tcagcggcgt
     3781 tttttctgct tttcttcgcg aattaattcc gcttcgcaac atgtgagcac cggtttattg
     3841 actaccggaa gcagtgtgac cgtgtgcttc tcaaatgcct gaggccagtt tgctcaggct
     3901 ctccccgtgg aggtaataat tgacgatatg atcagtgcac ggctaactaa gcggcctgct
     3961 gactttctcg ccgatcaaaa ggcattttgc tattaaggga ttgacgaggg cgtatctgcg
     4021 cagtaagatg cgccccgcat tggaaacctg atcatgtaga tcgaatggac tctaaatccg
     4081 ttcagccggg ttagattccc ggggtttccg ccaaattcga aaagcctgct caacgagcag
     4141 gcttttttgc atgctcgagc agctcagggt cgaatttgct ttcgaatttc tgccattcat
     4201 ccgcttatta tcacttattc aggcgtagca accaggcgtt taagggcacc aataactgcc
     4261 ttaaaaaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat
     4321 tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag
     4381 caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc
     4441 catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa
     4501 aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac
     4561 atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga
     4621 tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat
     4681 caccagctca ccgtctttca ttgccatacg gaattccgga tgagcattca tcaggcgggc
     4741 aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa
     4801 ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc
     4861 ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt
     4921 tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     4981 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca
     5041 ttttcgccaa aagttggccc agggcttccc ggtatcaaca gggacaccag gatttattta
     5101 ttctgcgaag tgatcttccg tcacaggtat ttattcggcg caaagtgcgt cgggtgatgc
     5161 tgccaactta ctgatttagt gtatgatggt gtttttgagg tgctccagtg gcttctgttt
     5221 ctatcagctg tccctcctgt tcagctactg acggggtggt gcgtaacggc aaaagcaccg
     5281 ccggacatca gcgctagcgg agtgtatact ggcttactat gttggcactg atgagggtgt
     5341 cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac cggtgcgtca gcagaatatg
     5401 tgatacagga tatattccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg
     5461 cggcgagcgg aaatggctta cgaacggggc ggagatttcc tggaagatgc caggaagata
     5521 cttaacaggg aagtgagagg gccgcggcaa agccgttttt ccataggctc cgcccccctg
     5581 acaagcatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca ggactataaa
     5641 gataccaggc gt
//
