LOCUS       40924_26381        4849 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pJW20, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4849)
  AUTHORS   Dean C, Barkan D, Bermingham A, Blais J, Casey F, Casarez A, Colvin 
            R, Fuller J, Jones A, Li C, Lopez S, Metzger L, Prathapam R, Rasper 
            D, Reck F, Ruzin A, Shaul J, Shen X, Simmons R, Skewes-Cox P, 
            Tamrakar P, Uehara T.
  TITLE     The monobactam LYS228: mode of action and mechanisms decreasing in 
            vitro susceptibility of Escherichia coli and Klebsiella pneumoniae
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4849)
  AUTHORS   Wei J-R., Mostafavi M, Takeoka K.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-JUL-2018) Infectious Diseases, Bacteriology, Novartis 
            Institutes for Biomedical Research, 5300 Chiron way, Emeryville, CA 
            94608, USA
REFERENCE   3  (bases 1 to 4849)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4849)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-JUL-2018) Infectious Diseases, Bacteriology, Novartis Institutes
            for Biomedical Research, 5300 Chiron way, Emeryville, CA 94608, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(83..1501)
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     promoter        1738..1766
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1955..2485
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     promoter        2794..2822
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     misc_RNA        2828..2847
                     /product="gRNA variable region"
     misc_RNA        2848..2923
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      2945..2979
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     rep_origin      3019..3474
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      3585..4173
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4359..4499)
                     /label=bom
                     /note="basis of mobility region from pBR322"
ORIGIN
        1 gttcatatgg gattcacctt tatgttgata agaaataaaa gaaaatgcca ataggatatc
       61 ggcattttct tttgcgtttt tatttgttaa ctgttaattg tccttgttca aggatgctgt
      121 ctttgacaac agatgttttc ttgcctttga tgttcagcag gaagctaggc gcaaacgttg
      181 attgtttgtc tgcgtagaat cctctgtttg tcatatagct tgtaatcacg acattgtttc
      241 ctttcgcttg aggtacagcg aagtgtgagt aagtaaaggt tacatcgtta ggatcaagat
      301 ccatttttaa cacaaggcca gttttgttca gcggcttgta tgggccagtt aaagaattag
      361 aaacataacc aagcatgtaa atatcgttag acgtaatgcc gtcaatcgtc atttttgatc
      421 cgcgggagtc agtgaacaga taccatttgc cgttcatttt aaagacgttc gcgcgttcaa
      481 tttcatctgt tactgtgtta gatgcaatca gcggtttcat cacttttttc agtgtgtaat
      541 catcgtttag ctcaatcata ccgagagcgc cgtttgctaa ctcagccgtg cgttttttat
      601 cgctttgcag aagtttttga ctttcttgac ggaagaatga tgtgcttttg ccatagtatg
      661 ctttgttaaa taaagattct tcgccttggt agccatcttc agttccagtg tttgcttcaa
      721 atactaagta tttgtggcct ttatcttcta cgtagtgagg atctctcagc gtatggttgt
      781 cgcctgagct gtagttgcct tcatcgatga actgctgtac attttgatac gtttttccgt
      841 caccgtcaaa gattgattta taatcctcta caccgttgat gttcaaagag ctgtctgatg
      901 ctgatacgtt aacttgtgca gttgtcagtg tttgtttgcc gtaatgttta ccggagaaat
      961 cagtgtagaa taaacggatt tttccgtcag atgtaaatgt ggctgaacct gaccattctt
     1021 gtgtttggtc ttttaggata gaatcatttg catcgaattt gtcgctgtct ttaaagacgc
     1081 ggccagcgtt tttccagctg tcaatagaag tttcgccgac tttttgatag aacatgtaaa
     1141 tcgatgtgtc atccgcattt ttaggatctc cggctaatgc aaagacgatg tggtagccgt
     1201 gatagtttgc gacagtgccg tcagcgtttt gtaatggcca gctgtcccaa acgtccaggc
     1261 cttttgcaga agagatattt ttaattgtgg acgaatcgaa ctcaggaact tgatattttt
     1321 catttttttg ctgttcaggg atttgcagca tatcatggcg tgtaatatgg gaaatgccgt
     1381 atgtttcctt atatggcttt tggttcgttt ctttcgcaaa cgcttgagtt gcgcctcctg
     1441 ccagcagtgc ggtagtaaag gttaatactg ttgcttgttt tgcaaacttt ttgatgttca
     1501 tcgttcatgt ctcctttttt atgtactgtg ttagcggtct gcttcttcca gccctcctgt
     1561 ttgaagatgg caagttagtt acgcacaata aaaaaagacc taaaatatgt aaggggtgac
     1621 gccaaagtat acactttgcc ctttacacat tttaggtctt gcctgcttta tcagtaacaa
     1681 acccgcgcga tttacttttc gacctcattc tattagactc tcgttaccga gctcgaattg
     1741 acataagcct gttcggttcg taaactgtaa tgcaagtagc gtatgcgctc acgcaactgg
     1801 tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt
     1861 tatgactgtt tttttgtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc
     1921 gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcagca acgatgttac
     1981 gcagcagggc agtcgcccta aaacaaagtt aggtggctca agtatgggca tcattcgcac
     2041 atgtaggctc ggccctgacc aagtcaaatc catgcgggct gctcttgatc ttttcggtcg
     2101 tgagttcgga gacgtagcca cctactccca acatcagccg gactccgatt acctcgggaa
     2161 cttgctccgt agtaagacat tcatcgcgct tgctgccttc gaccaagaag cggttgttgg
     2221 cgctctcgcg gcttacgttc tgcccaggtt tgagcagccg cgtagtgaga tctatatcta
     2281 tgatctcgca gtctccggcg agcaccggag gcagggcatt gccaccgcgc tcatcaatct
     2341 cctcaagcat gaggccaacg cgcttggtgc ttatgtgatc tacgtgcaag cagattacgg
     2401 tgacgatccc gcagtggctc tctatacaaa gttgggcata cgggaagaag tgatgcactt
     2461 tgatatcgac ccaagtaccg ccacctaaca attcgcctga attgactctc ttccgggcgc
     2521 tatcatgcca taccgcgaaa ggttttgcac cattcgatgg tgtcaacgta aatgccgctt
     2581 cgccttcgcg cgcgaattgc aagctgatcc gggcttatcg actgcacggt gcaccaatgc
     2641 ttctggcgtc aggcagccat cggaagctgt ggtatggctg tgcaggtcgt aaatcactgc
     2701 ataattcgtg tcgctcaagg cgcactcccg ttctggataa tgttttttgc gccgacatca
     2761 taacggttct ggcaaatatt ctgaaatgag ctgttgacaa ttaatcatcg gctcgtataa
     2821 tgtgtgggca acctggcata aaaaaccgtt ttagagctag aaatagcaag ttaaaataag
     2881 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgctttttac tccatctgga
     2941 tttgttcaga acgctcggtt gccgccgggc gttttttatg tcagaccaag tttactcata
     3001 tatactttag attgatttac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3061 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3121 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3181 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     3241 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3301 ccacgttctt taatagtgga ctcttgttcc aaacttgaac aacactcaac cctatctcgg
     3361 gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3421 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttaaaagg
     3481 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3541 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3601 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3661 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3721 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3781 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3841 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3901 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3961 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4021 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4081 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4141 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4201 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4261 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     4321 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     4381 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
     4441 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     4501 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     4561 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     4621 caccgaaacg cgcgaggcag caaggagatg gcgcccaaca gtcccccggc cacggggcct
     4681 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc
     4741 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
     4801 gccacgatgc gtccggcgta gaggatctgc tcatgtttga cagcttatc
//
