LOCUS       sequence-165559-        7014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Machinery plasmid expressing permissive pCNF synthetase and cognate 
            amber suppressing tRNA. Incorporates azidoPhenylalanine..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7014)
  AUTHORS   Miyake-Stoner SJ, Miller AM, Hammill JT, Peeler JC, Hess KR, Mehl 
            RA, Brewer SH
  TITLE     Probing protein folding using site-specifically encoded unnatural 
            amino acids as FRET donors with tryptophan.
  JOURNAL   Biochemistry. 2009 Jun 30;48(25):5953-62. doi: 10.1021/bi900426d.
  PUBMED    19492814
REFERENCE   2  (bases 1 to 7014)
  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"; journalName: 
            "Biochemistry"; date: "2009-06-30- 30"; volume: "48"; issue: "25"; 
            pages: "5953-62"
COMMENT     Sequence Label: pDule2-pCNF
FEATURES             Location/Qualifiers
     source          1..7014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1161..1190
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1206..1223
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1279..1296)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1449..1535
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1742..1759)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1876..2421)
                     /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(1956..1975)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2572..2663
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2664..3455
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     primer_bind     complement(3642..3661)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4787..4809
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4802..4818
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5802..5824
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5980..6858)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aactcagaat aagaaatgag gccgctcatg gcgttctgtt gcccgtctca ctggtgaaaa
       61 gaaaaacaac cctggcgccg cttctttgag cgaacgatca aaaataagtg gcgccccatc
      121 aaaaaaatat tctcaacata aaaaactttg tgtaatactt gtaacgctga gtttacgctt
      181 tgaggaatcc cccatggacg aatttgaaat gataaagaga aacacatctg aaattatcag
      241 cgaggaagag ttaagagagg ttttaaaaaa agatgaaaaa tctgctctga taggttttga
      301 accaagtggt aaaatacatt tagggcatta tctccaaata aaaaagatga ttgatttaca
      361 aaatgctgga tttgatataa ttatagtgtt ggctgattta catgcctatt taaaccagaa
      421 aggagagttg gatgagatta gaaaaatagg agattataac aaaaaagttt ttgaagcaat
      481 ggggttaaag gcaaaatatg tttatggaag tgaatggatg cttgataagg attatacact
      541 gaatgtctat agattggctt taaaaactac cttaaaaaga gcaagaagga gtatggaact
      601 tatagcaaga gaggatgaaa atccaaaggt tgctgaagtt atctatccaa taatgcaggt
      661 taatggtcct cattatcttg gcgttgatgt tgcagttgga gggatggagc agagaaaaat
      721 acacatgtta gcaagggagc ttttaccaaa aaaggttgtt tgtattcaca accctgtctt
      781 aacgggtttg gatggagaag gaaagatgag ttcttcaaaa gggaatttta tagctgttga
      841 tgactctcca gaagagatta gggctaagat aaagaaagca tactgcccag ctggagttgt
      901 tgaaggaaat ccaataatgg agatagctaa atacttcctt gaatatcctt taaccataaa
      961 aaggccagaa aaatttggtg gagatttgac agttaatagc tatgaggagt tagagagttt
     1021 atttaaaaat aaggaattgc atccaatgga tttaaaaaat gctgtagctg aagaacttat
     1081 aaagatttta gagccaatta gaaagagatt ataactgcag tttcaaacgg gtaccatatg
     1141 ggaattcgaa gcttgggccc gaacaaaaac tcatctcaga agaggatctg aatagcgccg
     1201 tcgaccatca tcatcatcat cattgagttt aaacggtctc cagcttggct gttttggcgg
     1261 atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa
     1321 acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga
     1381 agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg
     1441 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     1501 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagctgtc cctcctgttc
     1561 agctactgac ggggtggtgc gtaacggcaa aagcaccgcc ggacatcagc gctagcggag
     1621 tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct tcatgtggca
     1681 ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata tattccgctt
     1741 cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg
     1801 aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa gtgagagggc
     1861 cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg aaatctgacg
     1921 ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1981 cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca ttccgctgtt
     2041 atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca gttcgctcca
     2101 agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta tccggtaact
     2161 atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca gccactggta
     2221 attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca
     2281 agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt ggtagctcag
     2341 agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca agagattacg
     2401 cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata tttctagatt
     2461 tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg
     2521 taattctcat gtttgacagc ttatcatcga tgcgaccgag tgagctagct atttgtttat
     2581 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2641 aataatattg aaaaaggaag agtatgaggg aagcggtgat cgccgaagta tcgactcaac
     2701 tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt
     2761 tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta
     2821 cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa
     2881 cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc
     2941 acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc
     3001 agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta
     3061 tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac
     3121 tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat
     3181 ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca
     3241 tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa
     3301 tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg
     3361 gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg
     3421 tgaaaggcga gatcaccaag gtagtcggca aataatgtct aacaatgtct aacaattcgt
     3481 tcaagccgag gggccgcaag atccggccac gatgacccgg tcgtcggttc agggcagggt
     3541 cgttaaatag ccgcttatgt ctattgctgg tttaccgggg atcctctacg ccggacgcat
     3601 cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac
     3661 cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat
     3721 ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct
     3781 tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc
     3841 gcataaggga gagcgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg
     3901 gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact
     3961 cgtaggacag gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag
     4021 cgcgacgatg atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc
     4081 cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat
     4141 ggcggccgac gcgctgggct acgtcttgct ggcgttcgcg acgcgaggct ggatggcctt
     4201 ccccattatg attcttctcg cttccggcgg catcgggatg cccgcgttgc aggccatgct
     4261 gtccaggcag gtagatgacg accatcaggg acagcttcaa ggatcgctcg cggctcttac
     4321 cagcctaact tcgatcattg gaccgctgat cgtcacggcg atttatgccg cctcggcgag
     4381 cacatggaac gggttggcat ggattgtagg cgccgcccta taccttgtct gcctccccgc
     4441 gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga atggaagccg gcggcacctc
     4501 gctaacggat tcaccactcc aagaattgga gccaatcaat tcttgcggag aactgtgaat
     4561 gcgcaaacca acccttggca gaacatatcc atcgcgtccg ccatctccag cagccgcacg
     4621 cggcgcatct cgggctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc
     4681 aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg tctggcgaaa
     4741 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     4801 tgtaaaacga cggccagtgc caagcttaaa aaaaatcctt agctttcgct aaggatctgc
     4861 agtggtccgg cgggccggat ttgaaccagc gccatgcgga tttagagtcc gccgttctgc
     4921 cctgctgaac taccgccggg aattcagcgt tacaagtatt acacaaagtt ttttatgttg
     4981 agaatatttt tttgatgggg cgccacttat ttttgatcgt tcgctcaaag aagcggcgcc
     5041 agggttgttt ttcttttcac cagtgagacg ggcaacagaa cgccatgagc ggcctcattt
     5101 cttattctga gttacaacag tccgcaccgc tgccggtagc tccttccggt gggcgcgggg
     5161 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     5221 gccggcagcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     5281 agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg
     5341 aacacctaca tctgtattaa cgaagcgcta accgttttta tcatgctctg ggaggcagaa
     5401 taaatgatca tatcgtcaat tattacctcc acggggagag cctgagcaaa ctggcctcag
     5461 gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa
     5521 tagacataag cggctattta acgaccctgc cctgaaccga cgaccgggtc gaatttgctt
     5581 tcgaatttct gccattcatc cgcttattat cacttattca ggcgtagcaa ccaggcgttt
     5641 aagggcacca ataactgcct taaaaaaatt acgccccgcc ctgccactca tcgcagttga
     5701 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     5761 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     5821 gaggttttca ccgtcatcac cgaaacgcgc gaggcagcag atcaattcgc gcgcgaaggc
     5881 gaagcggcat gcataatgtg cctgtcaaat ggacgaagca gggattctgc aaaccctatg
     5941 ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat tatgacaact tgacggctac
     6001 atcattcact ttttcttcac aaccggcacg gaactcgctc gggctggccc cggtgcattt
     6061 tttaaatacc cgcgagaaat agagttgatc gtcaaaacca acattgcgac cgacggtggc
     6121 gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg
     6181 ccagcttaag acgctaatcc ctaactgctg gcggaaaaga tgtgacagac gcgacggcga
     6241 caagcaaaca tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct
     6301 gatgtactga caagcctcgc gtacccgatt atccatcggt ggatggagcg actcgttaat
     6361 cgcttccatg cgccgcagta acaattgctc aagcagattt atcgccagca gctccgaata
     6421 gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga aatgcggctg
     6481 gtgcgcttca tccgggcgaa agaaccccgt attggcaaat attgacggcc agttaagcca
     6541 ttcatgccag taggcgcgcg gacgaaagta aacccactgg tgataccatt cgcgagcctc
     6601 cggatgacga ccgtagtgat gaatctctcc tggcgggaac agcaaaatat cacccggtcg
     6661 gcaaacaaat tctcgtccct gatttttcac caccccctga ccgcgaatgg tgagattgag
     6721 aatataacct ttcattccca gcggtcggtc gataaaaaaa tcgagataac cgttggcctc
     6781 aatcggcgtt aaacccgcca ccagatgggc attaaacgag tatcccggca gcaggggatc
     6841 attttgcgct tcagccatac ttttcatact cccgccattc agagaagaaa ccaattgtcc
     6901 atattgcatc agacattgcc gtcactgcgt cttttactgg ctcttctcgc taaccaaacc
     6961 ggtaacccgg gaccaagttt actcatatat acggacagcg gtgcggactg ttgt
//
