LOCUS       Exported               10172 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pVK9PtacMuAB, complete sequence.
ACCESSION   KP272129
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10172)
  AUTHORS   Gorshkova NV, Gak ER, Tokmakova IL, Mashko SV.
  TITLE     Genetic tools C. glutamicum
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10172)
  AUTHORS   Gorshkova NV, Gak ER, Tokmakova IL, Mashko SV.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-DEC-2014) Lab3, AGRI, 1st Dorozhny Proezd, 1-1, Moscow
            117545, Russia
REFERENCE   3  (bases 1 to 10172)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10172
                     /organism="Cloning vector pVK9PtacMuAB"
                     /mol_type="other DNA"
                     /db_xref="taxon:1655359"
     CDS             complement(463..1857)
                     /codon_start=1
                     /gene="rep"
                     /product="Rep"
                     /label=rep
                     /note="replication protein of pCG1 plasmid from C. 
                     glutamicum"
                     /protein_id="AKN19584.1"
                     /translation="MTLADSRDIDTPAWKFSADLFDTHPELALRSRGWTSEDRREFLAH
                     LGRENFQGSKTRDFASAWIKDPDTGETQPKLYRVGSKSLARCQYVALTHAQHAAVLVLD
                     IDVPSHQAGGKIEHVNPEVYAILERWARLEKAPAWIGVNPLSGKCQLIWLIDPVYAAAG
                     MSSPNMRLLAATTEEMTRVFGADQAFSHRLSRWPLHVSDDPTAYRWHAQHNRVDRLADL
                     MEVARMISGTEKPKKRYEQEFSSGRARIEAARKATAEAKALATLEASLPSAAEASGELI
                     DGVRVLWTAPGRAARDETAFRHALTVGYQLKAAGERLKDTKIIDAYERAYTVAQAVGAD
                     GREPDLPPMRDRQTMARRVRGYVAKGQPVVPARQTETQSSRGRKALATMGRRGGKKAAE
                     RWKDPNSEYARAQREKLAKSSQRQARKAKGNRLTIAGWFMTVEGETGSWRQSMKLCLNL
                     ACHVRP"
     gene            complement(463..1857)
                     /gene="rep"
                     /label=rep
     CDS             complement(2996..3529)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     CDS             complement(2996..3460)
                     /codon_start=1
                     /gene="aacC1"
                     /product="AacC1(GmR)"
                     /label=aacC1
                     /note="aminoglycoside 3-N-acetyltransferase 
                     (AacC1)##aminoglycoside 3-N-acetyltransferase (AacC1) 
                     confers resistance to gentamicin####"
                     /protein_id="AKN19585.1"
                     /translation="MGIIRTCRLGPDQVKSMRAALDLFGREFGDVATYSQHQPDSDYLG
                     NLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRSEIYIYDLAVSGEHRRQGIATALI
                     NLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIREEVMHFDIDPSTAT"
     gene            complement(2996..3460)
                     /gene="aacC1"
                     /label=aacC1
     promoter        complement(3718..3746)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     primer_bind     4150..4166
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(4212..5294)
                     /codon_start=1
                     /gene="lacIq"
                     /product="LacIq"
                     /label=lacIq
                     /note="lactose-inducible transcriptional repressor of the 
                     lac operon (LacIq)"
                     /protein_id="AKN19586.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(4212..5294)
                     /gene="lacIq"
                     /label=lacIq
     promoter        complement(5295..5371)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     regulatory      5384..5448
                     /regulatory_class="promoter"
                     /note="Ptac"
     promoter        5390..5418
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    5426..5442
                     /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             5480..7471
                     /codon_start=1
                     /gene="muA"
                     /product="MuA"
                     /label=muA
                     /note="transposase(MuA)"
                     /protein_id="AKN19587.1"
                     /translation="MELWVSPKECANLPGLPKTSAGVIYVAKKQGWQNRTRAGVKGGKA
                     IEYNANSLPVEAKAALLLRQGEIETSLGYFEIARPTLEAHDYDREALWSKWDNASDSQR
                     RLAEKWLPAVQAADEMLNQGISTKTAFATVAGHYQVSASTLRDKYYQVQKFAKPDWAAA
                     LVDGRGASRRNVHKSEFDEDAWQFLIADYLRPEKPAFRKCYERLELAAREHGWSIPSRA
                     TAFRRIQQLDEAMVVACREGEHALMHLIPAQQRTVEHLDAMQWINGDGYLHNVFVRWFN
                     GDVIRPKTWFWQDVKTRKILGWRCDVSENIDSIRLSFMDVVTRYGIPEDFHITIDNTRG
                     AANKWLTGGAPNRYRFKVKEDDPKGLFLLMGAKMHWTSVVAGKGWGQAKPVERAFGVGG
                     LEEYVDKHPALAGAYTGPNPQAKPDNYGDRAVDAELFLKTLAEGVAMFNARTGRETEMC
                     GGKLSFDDVFEREYARTIVRKPTEEQKRMLLLPAEAVNVSRKGEFTLKVGGSLKGAKNV
                     YYNMALMNAGVKKVVVRFDPQQLHSTVYCYTLDGRFICEAECLAPVAFNDAAAGREYRR
                     RQKQLKSATKAAIKAQKQMDALEVAELLPQIAEPAAPESRIVGIFRPSGNTERVKNQER
                     DDEYETERDEYLNHSLDILEQNRRKKAI"
     gene            5480..7471
                     /gene="muA"
                     /label=muA
     CDS             7510..8448
                     /codon_start=1
                     /gene="muB"
                     /product="MuB"
                     /label=muB
                     /protein_id="AKN19588.1"
                     /translation="MNISDIRAGLRTLVENEETTFKQIALESGLSTGTISSFINDKYNG
                     DNERVSQMLQRWLEKYHAVAELPEPPRFVETQTVKQIWTSMRFASLTESIAVVCGNPGV
                     GKTEAAREYRRTNNNVWMITITPSCASVLECLTELAFELGMNDAPRRKGPLSRALRRRL
                     EGTQGLVIIDEADHLGAEVLEELRLLQESTRIGLVLMGNHRVYSNMTGGNRTVEFARLF
                     SRIAKRTAINKTKKADVKAIADAWQINGEKELELLQQIAQKPGALRILNHSLRLAAMTA
                     HGKGERVNEDYLRQAFRELDLDVDISTLLRN"
     gene            7510..8448
                     /gene="muB"
                     /label=muB
     primer_bind     complement(8943..8959)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8967..8983
                     /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)."
     promoter        complement(8991..9021)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9036..9057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(9527..10126)
                     /direction=LEFT
     rep_origin      complement(9529..10117)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"