LOCUS       Exported                9940 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTac-Pg6MSAS-sfp, complete sequence.
ACCESSION   MH488924
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9940)
  AUTHORS   Kim B, Binkley R, Kim HU, Lee SY.
  TITLE     Metabolic engineering of Escherichia coli for the enhanced 
            production of l-tyrosine
  JOURNAL   Biotechnol. Bioeng. (2018) In press
  PUBMED    30019750
REFERENCE   2  (bases 1 to 9940)
  AUTHORS   Yang D, Kim WJ, Yoo SM, Choi JH, Ha SH, Lee MH, Lee SY.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JUN-2018) Dept. Chemical and Biomolecular Engineering,
            Korea Advanced Institute of Science and Technology, Daehak-ro 291, 
            Daejeon 34141, South Korea
REFERENCE   3  (bases 1 to 9940)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9940
                     /organism="Cloning vector pTac-Pg6MSAS-sfp"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:2301159"
     CDS             9..5333
                     /codon_start=1
                     /gene="6MSAS"
                     /product="6-methylsalicylic acid synthase"
                     /label=6MSAS
                     /note="derived from Penicillium griseofulvum"
                     /protein_id="AXN69998.1"
                     /translation="MHSAATSTYPSGKTSPAPVGTPGTEYSEYEFSNDVAVVGMACRVA
                     GGNHNPELLWQSLLSQKSAMGEIPPMRWEPYYRRDARNEKFLKNTTSRGYFLDRLEDFD
                     CQFFGISPKEAEQMDPQQRVSLEVASEALEDAGIPAKSLSGSDTAVFWGVNSDDYSKLV
                     LEDLPNVEAWMGIGTAYCGVPNRISYHLNLMGPSTAVDAACASSLVAIHHGVQAIRLGE
                     SKVAIVGGVNALCGPGLTRVLDKAGAISSDGSCKSFDDDAHGYARGEGAGALVLKSLHR
                     ALLDHDNVLAVIKGSAVCQDGKTNGIMAPNSVAQQLAANNALSAANIDPHTVRYVEAHA
                     TSTPLGDPTEISAIASVYGADRPADDPCYIGSIKPNIGHLEAGAGVMGFIKAVLAIQKG
                     VLPPQANLTKLNSRIDWKTAGVKVVQEATPWPESDPIRRAGVCSYGYGGTVSHAVIEEF
                     SPILQPDPLGNGAVSGPGLLLLSGPQEKRLALQAKTLRDWMTAEGKDHNLSDILTTLAT
                     RRDHHDYRAALVVDDYRDAEQVLQSLANGVDHTFTTQSRVLGSDISKDVVWVFSGHGAQ
                     WPDMGKQLIHNPVFFAAIQPLDELIQAEIGLSPIELLRTGDFESSDRVQILTYVMQIGL
                     SALLQSNGITPQAVIGHSVGEIAASVVAGALSPAEGALIVTRRALLYRQVMGKGGMILV
                     NLPSAETEEILGSRSDLVVAIDSSPSSCVVAGDKELVAETAEALKARGVKTFTVKSDIA
                     FHSPTLNGLVDPLRDVLAETLSPVSPNVKLYSTALADPRGQDLRDVEYWAGNMVNRVRL
                     TSAVKAAVEDGYRLFLEVSTHPVVSHSINETLMDAGMEDFAVIPTLLRKKPTEKHILHS
                     IAQLHCRGAEVNWAAQMPGRWATGVPTTTWMHKPIWRKIETAPLHTGLTHDVEKHTLLG
                     QRIPVPGTDTYVYTTRLDNDTKPFPGSHPLHGTEIVPAAGLINTFLKGTGGQMLQNVVL
                     RVPVAINAPRSVQVVVQQDQVKVVSRLIPSEPSQLDDDASWVTHTTAYWDRKVAGSEDR
                     IDFAAVKSRLVTKLADNFSIDYLDKVGVSAMGFPWAVTEHYRNDKEMLARVDVNPAISG
                     DAPLPWDSSSWAPVLDAATSVGSTIFPTPALRMPAQIERVEVFTSQDPPKISWLYVQEA
                     SDSVPTSHVSVVSEAGEVLAKFTAMRFSEIEGTPGVSGSMESLVHQIAWPPATPAEEPL
                     SIETVILVSPDATTRALYAASLPTRVNSFQFSSTQEFFSNASSLPLEKGTVVTYIPGEV
                     ASLAEVPAASESFTWNLLELIKFTVNGSLPIKVFTLTANIGEGQTPTALAQSPLYGLAR
                     VIASEHPDLGTLIDVEEPVIPLSTMRYIQGADIIRINDGIARTSRFRSLPRNKLLPASE
                     GPRLLPRPEGTYLITGGLGVLGLEVADFLVEKGARRLLLISRRALPPRRTWDQVSEDLQ
                     PTIAKIRLLESRGASVHVLPLDITKPDAVEQLTTALDRLSLPSVQGVVHAAGVLDNELV
                     MQTTRDAFNRVLAPKIAGALALHEVFPPKSVDFFVMFSSCGNLVGFTGQASYGSGNAFL
                     DTLATHRARLGDAAVSFQWTSWRGLGMGASTDFINAELESKGITDVTRDEAFAAWQHLA
                     KYDMDHGVVLRSRAFEDGEPIPVSILNDIAVRRVGTVSNTSPAAAGSSDAVPTSGPELK
                     AYLDEKIRGCVAKVLQMTAEDVDSKAALADLGVDSVMTVTLRRQLQLTLKIAVPPTLTW
                     SHPTVSHLAVWFAEKLAK"
     gene            9..5333
                     /gene="6MSAS"
                     /label=6MSAS
     regulatory      5451..5479
                     /regulatory_class="promoter"
                     /note="Tac promoter"
     promoter        5451..5479
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    5487..5503
                     /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             5526..6200
                     /codon_start=1
                     /gene="sfp"
                     /product="4-phosphopantetheinyl transferase"
                     /label=sfp
                     /note="derived from Bacillus subtilis"
                     /protein_id="AXN69999.1"
                     /translation="MKIYGIYMDRPLSQEENERFMSFISPEKREKCRRFYHKEDAHRTL
                     LGDVLVRSVISRQYQLDKSDIRFSTQEYGKPCIPDLPDAHFNISHSGRWVICAFDSQPI
                     GIDIEKTKPISLEIAKRFFSKTEYSDLLAKDKDEQTDYFYHLWSMKESFIKQEGKGLSL
                     PLDSFSVRLHQDGQVSIELPDSHSPCYIKTYEVDPGYKMAVCAVHPDFPEDITMVSYEE
                     LL"
     gene            5526..6200
                     /gene="sfp"
                     /label=sfp
     misc_feature    6201..6257
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     regulatory      6255..6572
                     /regulatory_class="terminator"
                     /note="rrnBT1T2"
     terminator      6460..6546
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6638..6665
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        6685..6776
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7089..7105)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             7348..8163
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /protein_id="AXN70000.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      8692..9237
                     /direction=RIGHT
                     /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."
     rep_origin      8745..9195
                     /label=p15A
                     /note="p15A"
     regulatory      9874..9902
                     /regulatory_class="promoter"
                     /note="Tac promoter"
     promoter        9874..9902
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    9910..9926
                     /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)."