LOCUS       sequence-334781-        6125 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tRNA synthetase/tRNA pair for the in vivo incorporation of a 
            photocrosslinker, p-azido-l-phenylalanine, into proteins in E coli 
            response to the amber codon, TAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6125)
  AUTHORS   Chin JW, Santoro SW, Martin AB, King DS, Wang L, Schultz PG
  TITLE     Addition of p-azido-L-phenylalanine to the genetic code of 
            Escherichia coli.
  JOURNAL   J Am Chem Soc. 2002 Aug 7;124(31):9026-7.
  PUBMED    12148987
REFERENCE   2  (bases 1 to 6125)
  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: "J Am Chem 
            Soc."; date: "2002-08-7"; volume: "124(31)"; pages: "9026-7"
COMMENT     Sequence Label: pEVOL-pAzF
FEATURES             Location/Qualifiers
     source          1..6125
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             585..602
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(658..675)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(2578..3237)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3152..3171
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3238..3340)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3732..3749)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3866..4411)
                     /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(3946..3965)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4557..5435)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPSDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        5462..5746
                     /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     5669..5688
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 cgttgaatgt gtaccgtctg gcgctgaaaa ccacgctgaa acgcgcccgt cgttccatgg
       61 agctgattgc gcgcgaggat gagaatccaa aagttgctga ggttatttac cctattatgc
      121 aagttaatcc gttgcactac cagggtgttg atgttgccgt cggtggtatg gagcaacgca
      181 aaattcacat gctggcacgt gaactgctgc cgaaaaaggt tgtctgtatt cataatccgg
      241 tcctgaccgg cctggatggc gagggtaaaa tgagcagcag caagggtaac tttattgcag
      301 ttgacgatag cccggaagaa atccgtgcga agatcaagaa agcgtactgc ccggcaggcg
      361 tggttgaggg taacccgatc atggaaatcg ccaagtattt tctggaatac ccactgacga
      421 ttaagcgccc ggagaaattt ggcggcgacc tgaccgtcaa cagctacgag gagctggaaa
      481 gcttgtttaa gaacaaagaa ctgcatccga tgcgcctgaa aaacgccgtg gcggaagagc
      541 tgattaagat tctggaacca attcgcaaac gtctgtaagt cgaccatcat catcatcatc
      601 attgagttta aacggtctcc agcttggctg ttttggcgga tgagagaaga ttttcagcct
      661 gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag
      721 tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga
      781 tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa
      841 aggctcagtc gaaagactgg gccttgtttg tgagctcccg gtcatcaatc atccccataa
      901 tccttgttag attatcaatt ttaaaaaact aacagttgtc agcctgtccc gctttaatat
      961 catacgccgt tatacgttgt ttacgctttg aggaatccca tatggacgag ttcgaaatga
     1021 ttaaacgcaa caccagcgaa attatctctg aagaagagct gcgcgaggtg ctgaagaaag
     1081 acgagaagag cgcgactatt ggctttgagc cgtccggtaa aattcacctg ggtcactacc
     1141 tgcaaatcaa gaagatgatt gatctgcaaa acgctggttt tgacatcatt atcctgctgg
     1201 cggacctgca cgcctacctg aatcaaaagg gcgagctgga tgagattcgc aagatcggcg
     1261 actacaataa gaaagtcttc gaagccatgg gtttgaaggc taaatacgtc tacggtagca
     1321 attttcagct ggataaggat tacacgttga atgtgtaccg tctggcgctg aaaaccacgc
     1381 tgaaacgcgc ccgtcgttcc atggagctga ttgcgcgcga ggatgagaat ccaaaagttg
     1441 ctgaggttat ttaccctatt atgcaagtta atccgttgca ctaccagggt gttgatgttg
     1501 ccgtcggtgg tatggagcaa cgcaaaattc acatgctggc acgtgaactg ctgccgaaaa
     1561 aggttgtctg tattcataat ccggtcctga ccggcctgga tggcgagggt aaaatgagca
     1621 gcagcaaggg taactttatt gcagttgacg atagcccgga agaaatccgt gcgaagatca
     1681 agaaagcgta ctgcccggca ggcgtggttg agggtaaccc gatcatggaa atcgccaagt
     1741 attttctgga atacccactg acgattaagc gcccggagaa atttggcggc gacctgaccg
     1801 tcaacagcta cgaggagctg gaaagcttgt ttaagaacaa agaactgcat ccgatgcgcc
     1861 tgaaaaacgc cgtggcggaa gagctgatta agattctgga accaattcgc aaacgtctgt
     1921 aactgcagtt tcaaacgcta aattgcctga tgcgctacgc ttatcaggcc tacatgatct
     1981 ctgcaatata ttgagtttgc gtgcttttgt aggccggata aggcgttcac gccgcatccg
     2041 gcaagaaaca gcaaacaatc caaaacgccg cgttcagcgg cgttttttct gcttttcttc
     2101 gcgaattaat tccgcttcgc aacatgtgag caccggttta ttgactaccg gaagcagtgt
     2161 gaccgtgtgc ttctcaaatg cctgaggcca gtttgctcag gctctccccg tggaggtaat
     2221 aattgacgat atgatcagtg cacggctaac taagcggcct gctgactttc tcgccgatca
     2281 aaaggcattt tgctattaag ggattgacga gggcgtatct gcgcagtaag atgcgccccg
     2341 cattccggcg gtagttcagc agggcagaac ggcggactct aaatccgcat ggcaggggtt
     2401 caaatcccct ccgccggacc aaattcgaaa agcctgctca acgagcaggc ttttttgcat
     2461 gctcgagcag ctcagggtcg aatttgcttt cgaatttctg ccattcatcc gcttattatc
     2521 acttattcag gcgtagcaac caggcgttta agggcaccaa taactgcctt aaaaaaatta
     2581 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
     2641 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
     2701 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac
     2761 gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc
     2821 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata
     2881 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc
     2941 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc
     3001 gtctttcatt gccatacgga attccggatg agcattcatc aggcgggcaa gaatgtgaat
     3061 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc
     3121 cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc
     3181 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt
     3241 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     3301 ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccaaaa
     3361 gttggcccag ggcttcccgg tatcaacagg gacaccagga tttatttatt ctgcgaagtg
     3421 atcttccgtc acaggtattt attcggcgca aagtgcgtcg ggtgatgctg ccaacttact
     3481 gatttagtgt atgatggtgt ttttgaggtg ctccagtggc ttctgtttct atcagctgtc
     3541 cctcctgttc agctactgac ggggtggtgc gtaacggcaa aagcaccgcc ggacatcagc
     3601 gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct
     3661 tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata
     3721 tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa
     3781 atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa
     3841 gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg
     3901 aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt
     3961 ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca
     4021 ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca
     4081 gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta
     4141 tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca
     4201 gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac
     4261 tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt
     4321 ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca
     4381 agagattacg cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata
     4441 tttctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg
     4501 atataagttg taattctcat gtttgacagc ttatcatcga taagcttggt acccaattat
     4561 gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa ctcgctcggg
     4621 ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc aaaaccaaca
     4681 ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt cgcctggctg
     4741 atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg gaaaagatgt
     4801 gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg
     4861 tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc catcggtgga
     4921 tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag cagatttatc
     4981 gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg cccaaacagg
     5041 tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt
     5101 gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac ccactggtga
     5161 taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg cgggaacagc
     5221 aaaatatcac tcggtcggca aacaaattct cgtccctgat ttttcaccac cccctgaccg
     5281 cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat aaaaaaatcg
     5341 agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt aaacgagtat
     5401 cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc gccattcaga
     5461 gaagaaacca attgtccata ttgcatcaga cattgccgtc actgcgtctt ttactggctc
     5521 ttctcgctaa ccaaaccggt aaccccgctt attaaaagca ttctgtaaca aagcgggacc
     5581 aaagccatga caaaaacgcg taacaaaagt gtctataatc acggcagaaa agtccacatt
     5641 gattatttgc acggcgtcac actttgctat gccatagcat ttttatccat aagattagcg
     5701 gatcctacct gacgcttttt atcgcaactc tctactgttt ctccataccc gtttttttgg
     5761 gctaacagga ggaattagat ctatggacga gttcgaaatg attaaacgca acaccagcga
     5821 aattatctct gaagaagagc tgcgcgaggt gctgaagaaa gacgagaaga gcgcgactat
     5881 tggctttgag ccgtccggta aaattcacct gggtcactac ctgcaaatca agaagatgat
     5941 tgatctgcaa aacgctggtt ttgacatcat tatcctgctg gcggacctgc acgcctacct
     6001 gaatcaaaag ggcgagctgg atgagattcg caagatcggc gactacaata agaaagtctt
     6061 cgaagccatg ggtttgaagg ctaaatacgt ctacggtagc aattttcagc tggataagga
     6121 ttaca
//
