LOCUS       sequence-371914-        8395 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  biosynthesize sulfotyrosine in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8395)
  AUTHORS   Chen Y, Jin S, Zhang M, Hu Y, Wu KL, Chung A, Wang S, Tian Z, Wang 
            Y, Wolynes PG, Xiao H
  TITLE     Unleashing the potential of noncanonical amino acid biosynthesis to 
            create cells with precision tyrosine sulfation.
  JOURNAL   Nat Commun. 2022 Sep 16;13(1):5434. doi: 10.1038/s41467-022-33111-4.
  PUBMED    36114189
REFERENCE   2  (bases 1 to 8395)
  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: "Nat 
            Commun."; date: "2022-09-16"; volume: "13(1):5434. doi"; pages: " 
            10.1038/s41467-022-33111-4"
COMMENT     Sequence Label: pEvol-NnSULT1C1-cysDNCQ
FEATURES             Location/Qualifiers
     source          1..8395
                     /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             3464..3481
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3537..3554)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(7721..8380)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     8295..8314
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(8381..8395,1..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 aattagatct atggctctcg acaaaatgga ggacctttcc ttaaaagaaa ccgtcgtctc
     2581 gcgtgctgaa atctgcgaag tcgaaggtat cccgttcact aagcctattt gtagtacgtg
     2641 ggatcaagtg tggaaattca aagcccggcc tgatgactta ctcattgcga cttacacaaa
     2701 ggctggtacc acttggaccc aggagatcgt agatatgatt caacaaaatg gtgatgtaga
     2761 aaagtgccgt cgtgctacga cataccgccg gcacccgttc ctcgaatggt caatccagga
     2821 gccaccggct gcgagttatt ctgggcttga attagcggag gccatgccaa gcccacgtac
     2881 aattaaaact cacctgccgg tgcaactcct tccaccttcg ttctgggagc agaattgcaa
     2941 gatcatttat gtggcccgta atgcgaagga taacttagtg agttactatc actttcaccg
     3001 catgagcaaa gaaatgccag atcctgggac ctgggaggaa tttatggaaa aatttatgac
     3061 tgggaaagtc ttgtgggggt cctggtatga tcacgttaaa gggtggtgga aggctaagga
     3121 ccgtcaccgt atcctgtatt tgttttacga ggatatgaag gaaaatccta aacaagaaat
     3181 ccaaaaaatc ttgaaattcc tcgaaaagga tgtgaatcaa gaggttctga acaagattct
     3241 ccacaacacc tcttttgaga tcatgaagga caatccgatg actaactata caaccgagtt
     3301 tcagggcatc atggatcact cgatctcacc gtttatgcgt aagggcgtgg tgggcgactg
     3361 gaagaactac ttcacagtgg cacagaacga aaaattcgat gaggattaca aaaagaaaat
     3421 ggcagatacc tcgttggtct ttcgcactga gctgtgagtc gaccatcatc atcatcatca
     3481 ttgagtttaa acggtctcca gcttggctgt tttggcggat gagagaagat tttcagcctg
     3541 atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt
     3601 agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat
     3661 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
     3721 ggctcagtcg aaagactggg ccttgtttgt gagctcccgg tcatcaatca tccccataat
     3781 ccttgttaga ttatcaattt taaaaaacta acagttgtca gcctgtcccg ctttaatatc
     3841 atacgccgtt atacgttgtt tacgctttga ggaatcccat atggatcaaa tacgacttac
     3901 tcacctgcgg caactggagg cggaaagcat ccacattatt cgcgaggtgg cggcagaatt
     3961 ctcaaatccg gtgatgctct actctatcgg taaagattcc agcgtcatgc tgcatctggc
     4021 gcgcaaggcg ttttatccag gtacgctgcc tttcccgttg ctgcatgtcg ataccggctg
     4081 gaaattccgc gagatgtatg agttccgcga tcgtactgct aaagcctacg gctgcgaact
     4141 gctggtgcat aaaaacccgg aaggcgtggc gatggggatt aatccattcg tgcacggcag
     4201 cgcgaaacat accgatatta tgaaaactga aggcctgaaa caggcgctga acaaatacgg
     4261 ttttgatgcc gccttcggtg gtgcgcgccg tgacgaagag aaatcccgcg ctaaagagcg
     4321 aatttactct ttccgtgacc gcttccatcg ctgggatccg aaaaatcagc gcccggagct
     4381 gtggcacaac tacaacgggc aaattaacaa aggcgaaagc atccgcgtct tcccgctctc
     4441 taactggacc gagcaggata tctggcaata catctggctg gaaaatatcg acattgttcc
     4501 gctatatctc gctgcggaac gtccggttct ggaacgcgac ggtatgttga tgatgattga
     4561 tgacaaccgt atcgacctgc aaccgggcga agtgattaaa aaacggatgg tgcgtttccg
     4621 tacgctgggc tgctggccgc tgaccggtgc ggtggagtta aatgcacaaa cactgccgga
     4681 aatcatcgaa gagatgctgg tttccaccac cagtgaacgt cagggccgcg tgattgaccg
     4741 cgaccaggcg gggtctatgg agctgaaaaa acgtcagggg tatttttaag atgaacaccg
     4801 cacttgcaca acaaatcgcc aatgaaggcg gcgtcgaagc ctggatgatt gcgcaacaac
     4861 ataaaagcct gctgcgtttt ctgacctgtg gtagcgtcga tgacggcaaa agtactctga
     4921 ttggtcgtct gctgcacgat acccgccaaa tctacgaaga tcagctctca tcgctgcata
     4981 acgacagtaa gcgtcacggc acccagggcg aaaagctgga tctggctctg ctggtggacg
     5041 gcctgcaagc tgagcgcgaa cagggcatca ccattgacgt ggcctaccgc tatttctcta
     5101 ccgagaagcg taaatttatt atcgccgaca ccccagggca cgagcagtac acccgcaata
     5161 tggcgactgg cgcatcgaca tgtgaactgg cgatcttact gatcgatgcc cgtaaaggcg
     5221 tgctcgatca aacccgtcgt cacagtttta tctccacact gttggggatc aaacatctgg
     5281 tcgtggcgat caacaaaatg gatctggtgg attacagtga agagacgttc acccgtattc
     5341 gtgaagatta tttgaccttt gccgggcagc tgccgggtaa tctggatatc cgctttgtgc
     5401 cgctctctgc actggaaggc gacaacgtgg catcgcaaag tgaaagtatg ccgtggtaca
     5461 gcggtccgac actgctcgaa gtgctggaaa ccgtggagat ccagcgagtg gtggatgctc
     5521 agccaatgcg cttcccggtg cagtacgtta atcgcccgaa tctcgatttt cgtggttacg
     5581 ccggaacgct ggcatccggt cgcgtggaag tcgggcaacg tgtcaaagtg ctgccctctg
     5641 gtgtggaatc aaacgtcgcg cggatcgtga cttttgatgg tgatcgcgaa gaagcctttg
     5701 ccggagaagc gatcaccctg gtgctgacgg atgagatcga catcagccgt ggcgatctgc
     5761 tgctggcggc agacgaagcg ttaccggcgg tgcagagcgc gtcggtggat gtggtatgga
     5821 tggcggaaca gccgctttct ccagggcaga gttacgacat caaaattgcc ggtaagaaga
     5881 cgcgcgcgcg tgttgatggc attcgctatc aggttgatat taataacctt acccagcgtg
     5941 aagttgaaaa cctgccactg aatgggatcg gcctcgtgga tctcactttt gacgagccgc
     6001 tggtgttaga tcgttatcaa caaaatccgg tgacgggtgg gctgattttt atcgatcgcc
     6061 tgagcaatgt gaccgtgggt gccggtatgg tgcacgagcc agttagccag gcaactgctg
     6121 cgccatctga attcagtgca ttcgaactgg aattgaatgc tctggttcgt cgccactttc
     6181 cgcactgggg cgcgcgcgat ttgctggggg ataaataatg gcgctgcatg acgaaaacgt
     6241 cgtctggcat agccatccgg tcactgtgca acaacgcgag ctacaccacg gtcatcgtgg
     6301 tgtagtgctg tggtttaccg gcctctccgg gtccggtaaa tcaacggtcg ccggggcgct
     6361 ggaggaggcg ttacataaac tcggcgtcag tacgtatctg ctggatggcg acaatgttcg
     6421 ccacggatta tgcagcgatc tcggttttag cgatgccgat cgtaaagaga atatccgtcg
     6481 cgtcggtgaa gtggcgaatt tgatggttga agccggactg gtggtgctga ccgcatttat
     6541 ctcgccacac cgcgccgaac gccagatggt tcgcgaacgc gtaggagaag ggcgctttat
     6601 cgaagtgttt gtcgatacgc cgctggcgat ttgcgaagcc cgcgatccca aaggcttata
     6661 taagaaagcg cgtgccggtg aactgcgcaa ctttacggga atagattccg tttacgaagc
     6721 gcctgaatcg gcagaaattc atctcaatgg tgaacaatta gtaacaaatt tggtacagca
     6781 attattagat ctgttgagac agaacgatat tatcagatcc tgataagtta acaccgctca
     6841 cagagacgag gtggagaaat gttagatcaa gtatgccagc tcgcacggaa tgcaggcgat
     6901 gccattatgc aggtctacaa cgggacgaaa ccgatggacg tcgtcagcaa agcggacaat
     6961 tctccggtaa cggcagcgga tattgccgct cacaccgtta tcatggacgg tttacgtacg
     7021 ctgacaccgg atgttccggt cctttctgaa gaagatcctc ccggttggga agtccgtcag
     7081 cactggcagc gttactggct ggtagacccg ctggatggta ctaaagagtt tattaaacgt
     7141 aatggcgaat tcaccgttaa cattgcgctc attgaccatg gcaaaccgat tttaggcgtg
     7201 gtgtatgcgc cggtaatgaa cgtaatgtac agcgcggcag aaggcaaagc gtggaaagaa
     7261 gagtgcggtg tgcgcaagca gattcaggtc cgcgatgcgc gcccgccgct ggtggtgatc
     7321 agccgttccc atgcggatgc ggagctgaaa gagtatctgc aacagcttgg cgaacatcag
     7381 accacgtcca tcggctcttc gctgaaattc tgcctggtgg cggaaggaca ggcgcagctg
     7441 tacccgcgct tcggaccaac gaatatttgg gacaccgccg ctggacatgc tgtagctgca
     7501 gctgccggag cgcacgttca cgactggcag ggtaaaccgc tggattacac tccgcgtgag
     7561 tcgttcctga atccggggtt cagagtgtct atttactaag catgctcgag cagctcaggg
     7621 tcgaatttgc tttcgaattt ctgccattca tccgcttatt atcacttatt caggcgtagc
     7681 aaccaggcgt ttaagggcac caataactgc cttaaaaaaa ttacgccccg ccctgccact
     7741 catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg
     7801 catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc
     7861 ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg
     7921 tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga
     7981 aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc
     8041 ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa
     8101 cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac
     8161 ggaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact
     8221 tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt
     8281 tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg
     8341 atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagc
//
