LOCUS       Exported                7003 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Machinery plasmid expressing permissive pCNF synthetase and cognate 
            amber suppressing tRNA. Incorporates azidoPhenylalanine..
ACCESSION   .
VERSION     .
KEYWORDS    pDule2-pCNF
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7003)
  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 7003)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Biochemistry"; date: "2009-06-30- 30"; volume: "48"; issue: "25"; 
            pages: "5953-62"
FEATURES             Location/Qualifiers
     source          1..7003
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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(3631..3650)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4776..4798
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4790..4807
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4791..4807
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5791..5813
                     /label=pGEX 3
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5969..6847)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"