LOCUS       Exported                8181 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pNOV076, complete sequence.
ACCESSION   KX986298
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8181)
  AUTHORS   Jones AK, Woods AL, Takeoka KT, Shen X, Wei J-R., Caughlan RE, Dean 
            CR.
  TITLE     Determinants of antibacterial spectrum and resistance potential of 
            the elongation factor G inhibitor argyrin B in key Gram negative 
            pathogens
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8181)
  AUTHORS   Wei J-R.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-OCT-2016) Infectious Diseases, Bacteriology, Novartis 
            Institutes for Biomedical Research, 5300 Chiron Way, Emeryville, CA 
            94608, USA
REFERENCE   3  (bases 1 to 8181)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8181
                     /organism="Cloning vector pNOV076"
                     /mol_type="other DNA"
                     /db_xref="taxon:1931146"
     rep_origin      7..1337
     CDS             complement(1471..2562)
                     /codon_start=1
                     /gene="lacI"
                     /product="LacI"
                     /label=lacI
                     /note="lac operon repressor"
                     /protein_id="APS85741.1"
                     /translation="MVNVKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAM
                     AELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVER
                     SGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
                     FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGD
                     WSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTED
                     SSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTA
                     SPRALADSLMQLARQVSRLESGQ"
     gene            complement(1471..2562)
                     /gene="lacI"
                     /label=lacI
     CDS             complement(1471..2553)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(2554..2631)
                     /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."
     promoter        2861..2889
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2897..2913
                     /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             2936..5056
                     /codon_start=1
                     /gene="fusA"
                     /product="elongation factor G"
                     /label=fusA
                     /note="S459F"
                     /protein_id="APS85742.1"
                     /translation="MARTTPINRYRNIGICAHVDAGKTTTTERVLFYTGVNHKLGEVHD
                     GAATTDWMVQEQERGITITSAAVTTFWKGSRGQYDNYRVNVIDTPGHVDFTIEVERSLR
                     VLDGAVVVFCGTSGVEPQSETVWRQANKYGVPRIVYVNKMDRQGANFLRVVEQIKKRLG
                     HTPVPVQLAIGAEENFVGQVDLIKMKAIYWNDDDKGMTYREEEIPAELKDLAEEWRSSM
                     VEAAAEANEELMNKYLEEGELSEAEIKEGLRLRTLACEIVPAVCGSSFKNKGVPLVLDA
                     VIDYLPAPTEIPAIKGVSPDDETVEDERHADDNEPFSSLAFKIATDPFVGTLTFARVYS
                     GVLSSGDSVLNSVKGKKERVGRMVQMHANQREEIKEVRAGDIAALIGMKDVTTGDTLCS
                     IEKPIILERMDFPEPVISVAVEPKTKADQEKMGIALGKLAQEDPSFRVKTDEESGQTII
                     FGMGELHLDIIVDRMKREFGVEANIGKPQVAYRETITKDNVEIEGKFVRQSGGRGQFGH
                     CWIRFSAADVDEKGNITEGLVFENEVVGGVVPKEYIPAIQKGIEEQMKNGVVAGYPLIG
                     LKATVFDGSYHDVDSNEMAFKIAASMATKQLAQKGGGKVLEPIMKVEVVTPEDYMGDVM
                     GDLNRRRGLIQGMEDTVSGKVIRAEVPLGEMFGYATDVRSMSQGRASYSMEFSKYAEAP
                     SNIVEALVKKQG"
     gene            2936..5056
                     /gene="fusA"
                     /label=fusA
                     /note="S459F"
     primer_bind     complement(5066..5082)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5342..5428
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5520..5547
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5579..5607
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             5796..6329
                     /codon_start=1
                     /gene="accC1"
                     /product="AccC1"
                     /label=accC1
                     /note="gentamicin resistant protein"
                     /protein_id="APS85743.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            5796..6329
                     /gene="accC1"
                     /label=accC1
     rep_origin      6348..7509
     rep_origin      6351..6806
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      6917..7505
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    7691..7831
                     /label=bom
                     /note="basis of mobility region from pBR322"