LOCUS       sequence-418765-        6128 bp    DNA     circular SYN 14-APR-2025
DEFINITION  tRNA synthetase/tRNA pair for the in vivo incorporation of a 
            photocrosslinker, p-benzoyl-l-phenylalanine into proteins in E coli 
            in response to the amber codon, TAG. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6128)
  AUTHORS   Chin JW, Martin AB, King DS, Wang L, Schultz PG
  TITLE     Addition of a photocrosslinking amino acid to the genetic code of 
            Escherichiacoli.
  JOURNAL   Proc Natl Acad Sci U S A. 2002 Aug 20;99(17):11020-4. Epub 2002 Aug 
            1.
  PUBMED    12154230
REFERENCE   2  (bases 1 to 6128)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2002-08-20"; volume: "99(17)"; pages: 
            "11020-4. Epub 2002 Aug 1"
COMMENT     Sequence Label: pEVOL-pBpF
FEATURES             Location/Qualifiers
     source          1..6128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..306
                     /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     229..248
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1273..1290
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1346..1363)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(3266..3925)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3840..3859
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3926..4028)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4420..4437)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4554..5099)
                     /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(4634..4653)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5245..6123)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPSDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 tcatactccc gccattcaga gaagaaacca attgtccata ttgcatcaga cattgccgtc
       61 actgcgtctt ttactggctc ttctcgctaa ccaaaccggt aaccccgctt attaaaagca
      121 ttctgtaaca aagcgggacc aaagccatga caaaaacgcg taacaaaagt gtctataatc
      181 acggcagaaa agtccacatt gattatttgc acggcgtcac actttgctat gccatagcat
      241 ttttatccat aagattagcg gatcctacct gacgcttttt atcgcaactc tctactgttt
      301 ctccataccc gtttttttgg gctaacagga ggaattagat ctatggacga atttgaaatg
      361 ataaagagaa acacatctga aattatcagc gaggaagagt taagagaggt tttaaaaaaa
      421 gatgaaaaat ctgctggtat aggttttgaa ccaagtggta aaatacattt agggcattat
      481 ctccaaataa aaaagatgat tgatttacaa aatgctggat ttgatataat tatattgttg
      541 gctgatttac acgcctattt aaaccagaaa ggagagttgg atgagattag aaaaatagga
      601 gattataaca aaaaagtttt tgaagcaatg gggttaaagg caaaatatct ttatggaagt
      661 cctttccagc ttgataagga ttatacactg aatgtctata gattggcttt aaaaactacc
      721 ttaaaaagag caagaaggag tatggaactt atagcaagag aggatgaaaa tccaaaggtt
      781 gctgaagtta tctatccaat aatgcaggtt aatacgagtc attatctggg cgttgatgtt
      841 gcagttggag ggatggagca gagaaaaata cacatgttag caagggagct tttaccaaaa
      901 aaggttgttt gtattcacaa ccctgtctta acgggtttgg atggagaagg aaagatgagt
      961 tcttcaaaag ggaattttat agctgttgat gactctccag aagagattag ggctaagata
     1021 aagaaagcat actgcccagc tggagttgtt gaaggaaatc caataatgga gatagctaaa
     1081 tacttccttg aatatccttt aaccataaaa aggccagaaa aatttggtgg agatttgaca
     1141 gttaatagct atgaggagtt agagagttta tttaaaaata aggaattgca tccaatggat
     1201 ttaaaaaatg ctgtagctga agaacttata aagattttag agccaattag aaagagatta
     1261 taataagtcg accatcatca tcatcatcat tgagtttaaa cggtctccag cttggctgtt
     1321 ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc
     1381 tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga
     1441 actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag
     1501 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc cttgtttgtg
     1561 agctcccggt catcaatcat ccccataatc cttgttagat tatcaatttt aaaaaactaa
     1621 cagttgtcag cctgtcccgc tttaatatca tacgccgtta tacgttgttt acgctttgag
     1681 gaatcccata tggacgaatt tgaaatgata aagagaaaca catctgaaat tatcagcgag
     1741 gaagagttaa gagaggtttt aaaaaaagat gaaaaatctg ctggtatagg ttttgaacca
     1801 agtggtaaaa tacatttagg gcattatctc caaataaaaa agatgattga tttacaaaat
     1861 gctggatttg atataattat attgttggct gatttacacg cctatttaaa ccagaaagga
     1921 gagttggatg agattagaaa aataggagat tataacaaaa aagtttttga agcaatgggg
     1981 ttaaaggcaa aatatcttta tggaagtcct ttccagcttg ataaggatta tacactgaat
     2041 gtctatagat tggctttaaa aactacctta aaaagagcaa gaaggagtat ggaacttata
     2101 gcaagagagg atgaaaatcc aaaggttgct gaagttatct atccaataat gcaggttaat
     2161 acgagtcatt atctgggcgt tgatgttgca gttggaggga tggagcagag aaaaatacac
     2221 atgttagcaa gggagctttt accaaaaaag gttgtttgta ttcacaaccc tgtcttaacg
     2281 ggtttggatg gagaaggaaa gatgagttct tcaaaaggga attttatagc tgttgatgac
     2341 tctccagaag agattagggc taagataaag aaagcatact gcccagctgg agttgttgaa
     2401 ggaaatccaa taatggagat agctaaatac ttccttgaat atcctttaac cataaaaagg
     2461 ccagaaaaat ttggtggaga tttgacagtt aatagctatg aggagttaga gagtttattt
     2521 aaaaataagg aattgcatcc aatggattta aaaaatgctg tagctgaaga acttataaag
     2581 attttagagc caattagaaa gagattataa ctgcagtttc aaacgctaaa ttgcctgatg
     2641 cgctacgctt atcaggccta catgatctct gcaatatatt gagtttgcgt gcttttgtag
     2701 gccggataag gcgttcacgc cgcatccggc aagaaacagc aaacaatcca aaacgccgcg
     2761 ttcagcggcg ttttttctgc ttttcttcgc gaattaattc cgcttcgcaa catgtgagca
     2821 ccggtttatt gactaccgga agcagtgtga ccgtgtgctt ctcaaatgcc tgaggccagt
     2881 ttgctcaggc tctccccgtg gaggtaataa ttgacgatat gatcagtgca cggctaacta
     2941 agcggcctgc tgactttctc gccgatcaaa aggcattttg ctattaaggg attgacgagg
     3001 gcgtatctgc gcagtaagat gcgccccgca ttccggcggt agttcagcag ggcagaacgg
     3061 cggactctaa atccgcatgg caggggttca aatcccctcc gccggaccaa attcgaaaag
     3121 cctgctcaac gagcaggctt ttttgcatgc tcgagcagct cagggtcgaa tttgctttcg
     3181 aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaacca ggcgtttaag
     3241 ggcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg cagtactgtt
     3301 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
     3361 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg
     3421 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
     3481 gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
     3541 caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
     3601 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
     3661 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaat tccggatgag
     3721 cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct
     3781 ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag
     3841 caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg
     3901 tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact
     3961 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     4021 gccgatcaac gtctcatttt cgccaaaagt tggcccaggg cttcccggta tcaacaggga
     4081 caccaggatt tatttattct gcgaagtgat cttccgtcac aggtatttat tcggcgcaaa
     4141 gtgcgtcggg tgatgctgcc aacttactga tttagtgtat gatggtgttt ttgaggtgct
     4201 ccagtggctt ctgtttctat cagctgtccc tcctgttcag ctactgacgg ggtggtgcgt
     4261 aacggcaaaa gcaccgccgg acatcagcgc tagcggagtg tatactggct tactatgttg
     4321 gcactgatga gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt
     4381 gcgtcagcag aatatgtgat acaggatata ttccgcttcc tcgctcactg actcgctacg
     4441 ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa cggggcggag atttcctgga
     4501 agatgccagg aagatactta acagggaagt gagagggccg cggcaaagcc gtttttccat
     4561 aggctccgcc cccctgacaa gcatcacgaa atctgacgct caaatcagtg gtggcgaaac
     4621 ccgacaggac tataaagata ccaggcgttt ccccctggcg gctccctcgt gcgctctcct
     4681 gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc
     4741 acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta tgcacgaacc
     4801 ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccgga
     4861 aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag gagttagtct
     4921 tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac tgcgctcctc
     4981 caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa aaaccgccct
     5041 gcaaggcggt tttttcgttt tcagagcaag agattacgcg cagaccaaaa cgatctcaag
     5101 aagatcatct tattaatcag ataaaatatt tctagatttc agtgcaattt atctcttcaa
     5161 atgtagcacc tgaagtcagc cccatacgat ataagttgta attctcatgt ttgacagctt
     5221 atcatcgata agcttggtac ccaattatga caacttgacg gctacatcat tcactttttc
     5281 ttcacaaccg gcacggaact cgctcgggct ggccccggtg cattttttaa atacccgcga
     5341 gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag gcatccgggt
     5401 ggtgctcaaa agcagcttcg cctggctgat acgttggtcc tcgcgccagc ttaagacgct
     5461 aatccctaac tgctggcgga aaagatgtga cagacgcgac ggcgacaagc aaacatgctg
     5521 tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt actgacaagc
     5581 ctcgcgtacc cgattatcca tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg
     5641 cagtaacaat tgctcaagca gatttatcgc cagcagctcc gaatagcgcc cttccccttg
     5701 cccggcgtta atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg
     5761 gcgaaagaac cccgtattgg caaatattga cggccagtta agccattcat gccagtaggc
     5821 gcgcggacga aagtaaaccc actggtgata ccattcgcga gcctccggat gacgaccgta
     5881 gtgatgaatc tctcctggcg ggaacagcaa aatatcactc ggtcggcaaa caaattctcg
     5941 tccctgattt ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat aacctttcat
     6001 tcccagcggt cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc
     6061 cgccaccaga tgggcattaa acgagtatcc cggcagcagg ggatcatttt gcgcttcagc
     6121 catacttt
//
