LOCUS       Exported               10173 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pVK-lacIQ-Ptac-MuAB, complete sequence.
ACCESSION   MG014199
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10173)
  AUTHORS   Gorshkova NV, Lobanova JS, Tokmakova IL, Smirnov SV, Akhverdyan VZ, 
            Krylov AA, Mashko SV.
  TITLE     Mu-driven transposition of recombinant mini-Mu unit DNA in the 
            Corynebacterium glutamicum chromosome
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10173)
  AUTHORS   Gorshkova NV, Lobanova JS, Tokmakova IL, Smirnov SV, Akhverdyan VZ, 
            Krylov AA, Mashko SV.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-SEP-2017) Ajinomoto-Genetika Research Institute, 1st 
            Dorozhny pr. 1-1, Moscow 117545, Russian Federation
REFERENCE   3  (bases 1 to 10173)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..10173
                     /organism="Cloning vector pVK-lacIQ-Ptac-MuAB"
                     /mol_type="other DNA"
                     /db_xref="taxon:2049511"
     rep_origin      complement(404..1858)
                     /direction=LEFT
                     /label=ori_pCG1
                     /note="ori_pCG1"
     CDS             complement(2997..3530)
                     /codon_start=1
                     /product="gentamicin resistance protein"
                     /label=gentamicin resistance protein
                     /note="GmR"
                     /protein_id="ATO98418.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(3719..3747)
                     /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
                     /product="LacI"
                     /label=LacI
                     /protein_id="ATO98417.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     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      complement(5302..5371)
                     /regulatory_class="promoter"
                     /note="PlacIQ"
     regulatory      5361..5444
                     /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    5424..5444
                     /label=O lac binding site
                     /bound_moiety="O lac"
     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
                     /product="mu phage transposase subunit A"
                     /label=mu phage transposase subunit A
                     /note="MuA"
                     /protein_id="ATO98415.1"
                     /translation="MELWVSPKECANLPGLPKTSAGVIYVAKKQGWQNRTRAGVKGGKA
                     IEYNANSLPVEAKAALLLRQGEIETSLGYFEIARPTLEAHDYDREALWSKWDNASDSQR
                     RLAEKWLPAVQAADEMLNQGISTKTAFATVAGHYQVSASTLRDKYYQVQKFAKPDWAAA
                     LVDGRGASRRNVHKSEFDEDAWQFLIADYLRPEKPAFRKCYERLELAAREHGWSIPSRA
                     TAFRRIQQLDEAMVVACREGEHALMHLIPAQQRTVEHLDAMQWINGDGYLHNVFVRWFN
                     GDVIRPKTWFWQDVKTRKILGWRCDVSENIDSIRLSFMDVVTRYGIPEDFHITIDNTRG
                     AANKWLTGGAPNRYRFKVKEDDPKGLFLLMGAKMHWTSVVAGKGWGQAKPVERAFGVGG
                     LEEYVDKHPALAGAYTGPNPQAKPDNYGDRAVDAELFLKTLAEGVAMFNARTGRETEMC
                     GGKLSFDDVFEREYARTIVRKPTEEQKRMLLLPAEAVNVSRKGEFTLKVGGSLKGAKNV
                     YYNMALMNAGVKKVVVRFDPQQLHSTVYCYTLDGRFICEAECLAPVAFNDAAAGREYRR
                     RQKQLKSATKAAIKAQKQMDALEVAELLPQIAEPAAPESRIVGIFRPSGNTERVKNQER
                     DDEYETERDEYLNHSLDILEQNRRKKAI"
     CDS             7510..8448
                     /codon_start=1
                     /product="mu phage transposase subunit B"
                     /label=mu phage transposase subunit B
                     /note="MuB"
                     /protein_id="ATO98416.1"
                     /translation="MNISDIRAGLRTLVENEETTFKQIALESGLSTGTISSFINDKYNG
                     DNERVSQMLQRWLEKYHAVAELPEPPRFVETQTVKQIWTSMRFASLTESIAVVCGNPGV
                     GKTEAAREYRRTNNNVWMITITPSCASVLECLTELAFELGMNDAPRRKGPLSRALRRRL
                     EGTQGLVIIDEADHLGAEVLEELRLLQESTRIGLVLMGNHRVYSNMTGGNRTVEFARLF
                     SRIAKRTAINKTKKADVKAIADAWQINGEKELELLQQIAQKPGALRILNHSLRLAAMTA
                     HGKGERVNEDYLRQAFRELDLDVDISTLLRN"
     primer_bind     complement(8944..8960)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8968..8984
                     /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(8992..9022)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9037..9058
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(9530..10118)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(9531..10082)
                     /direction=LEFT