LOCUS       40924_33442        8181 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pNOV076, complete sequence.
ACCESSION   .
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  JOURNAL   Unpublished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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8181)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-OCT-2016) Infectious Diseases, Bacteriology, Novartis Institutes
            for Biomedical Research, 5300 Chiron Way, Emeryville, CA 94608, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..1337
     CDS             complement(1474..2553)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(2554..2631)
                     /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
                     /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
                     /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
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             5796..6326
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     rep_origin      6351..6806
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      6917..7505
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(7691..7831)
                     /label=bom
                     /note="basis of mobility region from pBR322"
ORIGIN
        1 atcgatcaac cgctctgtag gcttgaagaa cgcaacccta tagcagctat tgaaattgat
       61 gatttaattg aagaaaagac agatttactt gttgataatc gactgatggg gcgcacaggc
      121 agacagaaag atactaggga gttagtgata catccgcatt atgtggttgt atatgacatc
      181 actgatataa tacggatact cagagtgcta cacacatcgc atgagtggtc atgacttact
      241 catgtacttt ggattattta gtgttataaa atcctgattt ataaattttt ttttgttaaa
      301 aaagataaaa gccccttgca attgcttggg gctttaccgt aatttatggg gtacagatct
      361 tcgatactga catatcggca atcgaaagca ttaaggtttg acgaccgcta atgatttcac
      421 cacaggggct taatgtacct gtcttaaatt ctaaggtttt aactcgcttt gtcaagcata
      481 gaccccaaaa atttagccaa tgtctgtaac tcaatctgtc catgtgtggg tgatgaggta
      541 cagtgacgct agcacacatc ggaaaaacgc tattactagg ggaactgaac agagtagcgg
      601 acgcaatgag tagtcattta attggcggtt atgagcgtgt tcaggcggtg ctatcaatcg
      661 taatcataac agtggcagct tgatacagtg atgtcatccc tgatgcgaaa gcgaccgacc
      721 gacggtacat cgaatgggaa tactttaggg tgatttttaa gaatcgctct agggtgagta
      781 tttcccattc agctctgctc cctccctctg gtactttaat caaaagcact actaaacata
      841 tgtttttaaa taaaaaatat tgatatagag ataatattag taagaataat taaacaattg
      901 aatatagata aatcattgtt aaataaagat taattattaa aatgaatgta tacttatata
      961 taaatcaatg atttaaaata tttgataaag aaaacttttc aaaaaaaata taattgagat
     1021 tgtgtcattt cggtcaattc ttaatatgtt ccacgcaagt tttagctatg gtgctaaaca
     1081 gaaatttgct gaaaaagaac ttttcactga actggttaaa atgtaagcag cctgagagcc
     1141 gccaaaaatt ttaaaaacaa accgccttaa tcatcttcaa aaaatacctc taaaacctca
     1201 ccatttgcgt tttaagaccc atatttcatc ctgcccttat gttcccatgc tgatagctat
     1261 aaagtgtctg taatcgcttc ctatgacgtt ctaggctgtt gataactttt ggaacaacgc
     1321 cgaaacaggc tcaccctgat gcataatgtg cctgtcaaat ggacgaagca gggattctgc
     1381 aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt cgttaccaac agcttgcaat
     1441 tcgcgctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1501 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1561 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     1621 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     1681 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     1741 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     1801 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     1861 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     1921 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     1981 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     2041 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     2101 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     2161 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     2221 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     2281 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     2341 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     2401 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     2461 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     2521 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     2581 cgggcgctat catgccatac cgcgaaaggt tttgcaccat tcgatggtgt caacgtaaat
     2641 gccgcttcgc cttcgcgcgc gaattgcaag ctgatccggg cttatcgact gcacggtgca
     2701 ccaatgcttc tggcgtcagg cagccatcgg aagctgtggt atggctgtgc aggtcgtaaa
     2761 tcactgcata attcgtgtcg ctcaaggcgc actcccgttc tggataatgt tttttgcgcc
     2821 gacatcataa cggttctggc aaatattctg aaatgagctg ttgacaatta atcatcggct
     2881 cgtataatgt gtggaattgt gagcggataa caatttcaca caggaaacag aattcatggc
     2941 ccgtactaca cccatcaacc gctaccgtaa tatcggtatc tgtgcccacg ttgacgcggg
     3001 caagaccacc accacggagc gggttctgtt ctataccggc gtcaaccaca agctgggcga
     3061 agtgcatgac ggcgctgcga ccaccgactg gatggtgcag gagcaggagc gcggtatcac
     3121 cattacctcc gctgccgtta ccaccttctg gaagggttcg cgcggtcagt acgacaacta
     3181 tcgcgtaaac gtcatcgaca ctcccggcca cgtagacttc accatcgagg tggagcgttc
     3241 cctgcgtgta ctggacggcg ctgtcgtggt gttctgtggc acctccggcg tagagccgca
     3301 gtcggaaacc gtatggcgtc aggccaacaa gtacggcgta ccgcgcatcg tctacgtcaa
     3361 caagatggac cgccagggcg cgaacttcct acgcgtcgtc gagcagatca agaagcgcct
     3421 gggtcatacc ccggttccgg tccagctcgc catcggcgct gaagagaact tcgtcggtca
     3481 ggtcgacctg atcaagatga aggccatcta ctggaacgat gacgacaagg gcatgaccta
     3541 ccgcgaggaa gaaattcctg cggagctcaa ggacctggcc gaagagtggc gttccagcat
     3601 ggtcgaggct gcggccgaag ccaacgaaga gctgatgaac aagtacctcg aagagggcga
     3661 gctctccgaa gccgagatca aggaaggcct gcgtctgcgc accctggcct gcgagatcgt
     3721 tccggccgtc tgcggttcct cgttcaagaa caagggcgtt ccgctggttc tcgatgccgt
     3781 catcgactac ctcccggctc cgaccgagat ccctgcgatc aagggcgtca gcccggacga
     3841 cgagaccgtc gaagacgagc gtcatgccga cgacaacgaa ccgttctcgt ccctggcgtt
     3901 caagatcgct accgacccgt tcgtcggtac cctgaccttc gcccgcgttt actccggcgt
     3961 cctgagttcc ggcgactccg tcctgaactc ggtgaaaggc aagaaagagc gcgtcggtcg
     4021 gatggtgcag atgcacgcca accagcgtga agagatcaag gaagtgcgcg caggcgacat
     4081 cgctgccctg atcggcatga aggacgtcac caccggtgac accctgtgct cgatcgagaa
     4141 gccgatcatc ctcgagcgca tggacttccc cgagccggta atctcggtcg ctgtagagcc
     4201 gaagaccaag gctgaccagg agaagatggg tatcgccctc ggcaagctgg cccaggaaga
     4261 cccgtcgttc cgtgtcaaga ccgacgaaga gtccggtcag accatcatct tcggcatggg
     4321 cgagctgcac ctggacatca tcgtcgaccg catgaagcgc gagttcggcg tcgaggccaa
     4381 catcggcaag ccgcaggttg cctaccgtga gaccatcacc aaggacaacg tggagatcga
     4441 aggcaagttc gttcgccagt ccggtggtcg cggtcagttc ggtcattgct ggatccgttt
     4501 ctcggccgcg gacgtggacg agaagggcaa tatcaccgaa ggcctggtgt tcgagaacga
     4561 agtcgtgggc ggtgtggttc cgaaggaata catcccggcg atccagaagg gtatcgaaga
     4621 gcagatgaag aacggcgtcg ttgccggcta tccgctcatc ggcctgaagg ctaccgtttt
     4681 cgatggttcc taccatgacg tcgactccaa cgagatggcg ttcaagatcg ctgcttccat
     4741 ggcgaccaag cagttggccc agaagggcgg cggcaaggtg ctcgagccga tcatgaaggt
     4801 agaggtagtg actcctgagg actacatggg tgacgtgatg ggcgacctga accgtcgtcg
     4861 tggtctgatt caggggatgg aggacacggt ttccggtaag gttattcgtg ccgaggttcc
     4921 gctgggtgag atgttcggtt atgcgaccga cgtgcgttcc atgtcccagg gtcgcgcgag
     4981 ctactccatg gaattctcca aatacgccga agctccgtcg aacatcgtcg aagcgctggt
     5041 taaaaaacaa ggttgacatt cgtacgatcc actagttcta gagcggccgc caccgcggtg
     5101 gagctccagc ttttgttccc tttagaggca tgcaagcttg gctgttttgg cggatgagag
     5161 aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat
     5221 ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa
     5281 cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
     5341 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     5401 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     5461 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     5521 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttaccg agctcgaatt
     5581 gacataagcc tgttcggttc gtaaactgta atgcaagtag cgtatgcgct cacgcaactg
     5641 gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg
     5701 ttatgactgt ttttttgtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc
     5761 cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagc aacgatgtta
     5821 cgcagcaggg cagtcgccct aaaacaaagt taggtggctc aagtatgggc atcattcgca
     5881 catgtaggct cggccctgac caagtcaaat ccatgcgggc tgctcttgat cttttcggtc
     5941 gtgagttcgg agacgtagcc acctactccc aacatcagcc ggactccgat tacctcggga
     6001 acttgctccg tagtaagaca ttcatcgcgc ttgctgcctt cgaccaagaa gcggttgttg
     6061 gcgctctcgc ggcttacgtt ctgcccaggt ttgagcagcc gcgtagtgag atctatatct
     6121 atgatctcgc agtctccggc gagcaccgga ggcagggcat tgccaccgcg ctcatcaatc
     6181 tcctcaagca tgaggccaac gcgcttggtg cttatgtgat ctacgtgcaa gcagattacg
     6241 gtgacgatcc cgcagtggct ctctatacaa agttgggcat acgggaagaa gtgatgcact
     6301 ttgatatcga cccaagtacc gccacctaac aattcgcttt agattgattt acgcgccctg
     6361 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     6421 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     6481 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     6541 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     6601 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     6661 ccaaacttga acaacactca accctatctc gggctattct tttgatttat aagggatttt
     6721 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     6781 taacaaaata ttaacgttta caatttaaaa ggatctaggt gaagatcctt tttgataatc
     6841 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6901 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6961 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7021 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     7081 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7141 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7201 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7261 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7321 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7381 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7441 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7501 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7561 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     7621 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     7681 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     7741 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc
     7801 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
     7861 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     7921 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agcaaggaga
     7981 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     8041 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     8101 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatct
     8161 gctcatgttt gacagcttat c
//
