LOCUS       sequence-390718-        5108 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express the azidophenylalanine-specific variant of 
            Methanocaldococcus jannaschii TyrRS and its cognate amber suppressor
            tRNA and allow UAG codon to be translated into azidophenylalanine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5108)
  AUTHORS   Yamaguchi A, Matsuda T, Ohtake K, Yanagisawa T, Yokoyama S, Fujiwara
            Y, Watanabe T, Hohsaka T, Sakamoto K
  TITLE     Incorporation of a Doubly Functionalized Synthetic Amino Acid into 
            Proteins for Creating Chemical and Light-Induced Conjugates.
  JOURNAL   Bioconjug Chem. 2016 Jan 20;27(1):198-206. doi: 
            10.1021/acs.bioconjchem.5b00602. Epub 2015 Dec 16.
  PUBMED    26625213
REFERENCE   2  (bases 1 to 5108)
  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"; doi: 
            "10.1021/acs.bioconjchem.5b00602"; journalName: "Bioconjug Chem"; 
            date: "2016-01-20- 20"; volume: "27"; issue: "1"; pages: "198-206"
COMMENT     Sequence Label: pCDF-Az
FEATURES             Location/Qualifiers
     source          1..5108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             77..94
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             113..127
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     primer_bind     complement(492..511)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             503..1318
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      complement(1459..2197)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     2689..2708
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3886..4968)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4930..4949
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4969..5046)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(5092..5108,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(5092..5108,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggcacatc accaccacca tcacgtgggt accggttcga atgatgacga
      121 cgacaagagt ccggatccca attgggagct cgtgtacacg gcgcgcctgc aggtcgacaa
      181 gcttcctcgt gaagaaggtg ttgctgactc ataccaggcc tgaatcgccc catcatccag
      241 ccagaaagtg agggagccac ggttgatgag agctttgttg taggtggacc agttggtgat
      301 tttgaacttt tgctttgcca cggaacggtc tgcgttgtcg ggaagatgcg tgatctgatc
      361 cttcaactca gcaaaagttc gatttattca acaaagccac gttgtgtctc aaaatctctg
      421 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
      481 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctcgaagcc
      541 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gtgataatgt
      601 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt
      661 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
      721 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
      781 tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata
      841 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
      901 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
      961 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     1021 gcctgttgaa caagtctgga aagaaatgca taagctcttg ccattctcac cggattcagt
     1081 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     1141 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     1201 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     1261 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     1321 agaattggtt aattggttgt aacactttgg aaatttttga tccttagcga aagctaagga
     1381 ttttttttat cgcgacgcga ggctggatgg cctgctagct cactcggtcg ctacgctccg
     1441 ggcgtgagac tgcggcgggc gctgcggaca catacaaagt tacccacaga ttccgtggat
     1501 aagcagggga ctaacatgtg aggcaaaaca gcagggccgc gccggtggcg tttttccata
     1561 ggctccgccc tcctgccaga gttcacataa acagacgctt ttccggtgca tctgtgggag
     1621 ccgtgaggct caaccatgaa tctgacagta cgggcgaaac ccgacaggac ttaaagatcc
     1681 ccaccgtttc cggcgggtcg ctccctcttg cgctctcctg ttccgaccct gccgtttacc
     1741 ggatacctgt tccgcctttc tcccttacgg gaagtgtggc gctttctcat agctcacaca
     1801 ctggtatctc ggctcggtgt aggtcgttcg ctccaagctg ggctgtaagc aagaactccc
     1861 cgttcagccc gactgctgcg ccttatccgg taactgttca cttgagtcca acccggaaaa
     1921 gcacggtaaa acgccactgg cagcagccat tggtaactgg gagttcgcag aggatttgtt
     1981 tagctaaaca cgcggttgct cttgaagtgt gcgccaaagt ccggctacac tggaaggaca
     2041 gatttggttg ctgtgctctg cgaaagccag ttaccacggt taagcagttc cccaactgac
     2101 ttaaccttcg atcaaaccac ctccccaggt ggttttttcg tttacagggc aaaagattac
     2161 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctactgaac cgctctagat
     2221 ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt
     2281 gtaattctca tgttagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
     2341 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga aggctctcaa gggcatcggt
     2401 cgactaaaaa aaatccttag ctttcgctaa ggatcaaaaa tttccaatct gtgataaatg
     2461 gtcccgccta cgggatttga acccgtgcca tgcggattta gagtccgccg ttctgccctc
     2521 tgaactaagg cgggaatgcg tctgatggca gcgttacaag tattacacaa agttttttat
     2581 gttgagaata tttttttgat ggggcgccac ttatttttga tcgttcgctc aaagaagcgg
     2641 cgccgcatgc ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     2701 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     2761 gtagaggatc cagatcttat aatctctttc taattggctc taaaatcttt ataagttctt
     2821 cagctacagc attttttaaa cgcattggat gcaattcctt atttttaaat aaactctcta
     2881 actcctcata gctattaact gtcaaatctc caccaaattt ttctggcctt tttatggtta
     2941 aaggatattc aaggaagtat ttagctatct ccattattgg atttccttca acaactccag
     3001 ctgggcagta tgctttcttt atcttagccc taatctcttc tggagagtca tcaacagcta
     3061 taaaattccc ttttgaagaa ctcatctttc cttctccatc caaacccgtt aagacagggt
     3121 tgtgaataca aacaaccttt tttggtaaaa gctcccttgc taacatgtgt atttttctct
     3181 gctccatccc tccaactgca acatcaacgc cttgataatg tagaggatta acctgcatta
     3241 ttggatagat aacttcagca acctttggat tttcatcctc tcttgctata agttccatac
     3301 tccttcttgc tctttttaag gtagttttta aagccaatct atagacattc agtgtataat
     3361 ccttatcaag ctggaaatta cttccataaa catattttgc ctttaacccc attgcttcaa
     3421 aaactttttt gttataatct cctatttttc taatctcatc caactctcct ttctggttta
     3481 aataggcgtg taaatcagcc aacaatataa ttatatcaaa tccagcattt tgtaaatcaa
     3541 tcatcttttt tatttggaga taatgcccta aatgtatttt accacttggt tcaaaaccta
     3601 tggtagcaga tttttcatct ttttttaaaa cctctcttaa ctcttcctcg ctgataattt
     3661 cagatgtgtt tctctttatc atttcaaatt cgtccatatg aatctccaag tatataacga
     3721 ctgcaccttt gccggtacac gacttttcgc cagatgcgaa agagacatag aataaagtgc
     3781 cagaatcagg agtaccagcg attaaagcaa gatttttgca tcttttcagg gtgcaagacg
     3841 atcaatcttc cacgcatcag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     3901 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3961 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     4021 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     4081 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     4141 ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat
     4201 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     4261 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     4321 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     4381 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     4441 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     4501 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     4561 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     4621 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     4681 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     4741 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     4801 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     4861 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     4921 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     4981 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     5041 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaaa ttaatacgac
     5101 tcactata
//
