LOCUS       40924_29311        9064 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLW18, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9064)
  AUTHORS   Delaney NF, Kaczmarek ME, Ward LM, Swanson PK, Lee MC, Marx CJ.
  TITLE     Development of an Optimized Medium, Strain and High-Throughput 
            Culturing Methods for Methylobacterium extorquens
  JOURNAL   PLoS ONE 8 (4), E62957 (2013)
  PUBMED    23646164
REFERENCE   2  (bases 1 to 9064)
  AUTHORS   Delaney NF.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-MAR-2013) Organismic and Evolutionary Biology, Harvard
            University, 16 Divinity Ave., Cambridge, MA 02138, USA
REFERENCE   3  (bases 1 to 9064)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9064)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2013"; volume: "8"; issue: "4"; pages: "E62957"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-MAR-2013) Organismic and Evolutionary Biology, Harvard 
            University, 16 Divinity Ave., Cambridge, MA 02138, 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..9064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(4..861)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(862..966)
                     /label=AmpR promoter
     misc_feature    997..1548
                     /label=region upstream of celA gene
                     /note="region upstream of celA gene"
     misc_feature    1549..2075
                     /label=region downstream of celC gene
                     /note="region downstream of celC gene"
     CDS             complement(2679..3047)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(3080..3189)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     promoter        3826..3854
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             3902..5089
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     CDS             complement(5273..5929)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             complement(6030..7448)
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     promoter        complement(7449..7894)
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     rep_origin      complement(8306..8894)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
       61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
      121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
      181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
      661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
      721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
      781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
      841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
      901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
      961 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgct cctactcgac caagttcaac
     1021 gcctactatt cccgtttccg cgtcggctac atggtcgccg acggcgtcta tatcggcccg
     1081 gaggcgctgt tcctcggcga cgacttcttc cgccagtacc gcgtcggcgc ccatctctcg
     1141 ggcctgagct tcggccccgt ccagatgtcg ctcgcggccg gctacgtgcg ggaccgcgtc
     1201 cagggcacgg gctactattc cagcatcgag gcacgcgcga acttttagag cgcattccga
     1261 cggcgagcgg gccgccggtc gcggcggcgg ccgaagtcaa gcgcttggat caatgcggac
     1321 aatccgccgc cttgtctccg cgtctcacgg caaagtactg ctgttccaag cgaagaatgg
     1381 ccttcatatc aggaagacat tcctcgccat cgcatgtcat ttccgcgaaa ttcctgccga
     1441 atcttcggac attttctacg gatgacgcta cgctctgcat cgcaaaatca accttgttcg
     1501 cagagcggga cggcggcgtc gtgaacgcat ccggaaatcg agatctcata tgcacgaagt
     1561 gaggacgcct tcgatctcta caagtcggtc ctcgatggtc ctgccgatgc aggcgcgcgg
     1621 ctcgccacga tccagaaggc gatgagccat ctcaagatgg atcaggccga gcggctcatc
     1681 gccatgggac gcaccaaccc ggagggcgct tcggaattcg acgcgatccg cctcgacatc
     1741 acccgcgccc gcatcgccgc cttcctgcac gacgagccgg cccagatgcc gaccctggcc
     1801 gacctcaccg ccttccagac ctatgcccgc aagagcggcg acccgagcca gaccggcctg
     1861 ctcggctggt acgcctacaa gcggcgccag ttccgcgagg cgctggaatg gttcaagctc
     1921 gccatcggcc gcggcggcga cgccatggtc gcccacggcc tcgcccatac cctgcgcgaa
     1981 ctgaacatgc tgcgcgaggc ggaggaggtc gcctatgcct ggcgcgaccg cttcgtcggc
     2041 aacgagctgc tgttcatcga cgtgctcgaa gagctcacta gaggatccag ccgaccaggc
     2101 tttccacgcc cgcgtgccgc tccatgtcgt tcgcgcggtt ctcggaaacg cgctgccgcg
     2161 tttcgtgatt gtcacgctca agcccgtagt cccgttcgag cgtcgcgcag aggtcagcga
     2221 gggcgcggta ggcccgatac ggctcatgga tggtgtttcg ggtcgggtga atcttgttga
     2281 tggcgatatg gatgtgcagg ttgtcggtgt cgtgatgcac ggcactgacg cgctgatgct
     2341 cggcgaagcc aagcccagcg cagatgcggt cctcaatcgc gcgcaacgtc tccgcgtcgg
     2401 gcttctctcc cgcgcggaag ctaaccagca ggtgataggt cttgtcggcc tcggaacggg
     2461 tgttgccgtg ctgggtcgcc atcacctcgg ccatgacagc gggcagggtg tttgcctcgc
     2521 agttcgtgac gcgcacgtga cccaggcgct cggtcttgcc ttgctcgtcg gtgatgtact
     2581 tcaccagctc cgcgaagtcg ctcttcttga tggagcgcat ggggacgtgc ttggcaatca
     2641 cgcgcacccc ccggccgttt tagcggctaa aaaagtcatg gctctgccct cgggcggacc
     2701 acgcccatca tgaccttgcc aagctcgtcc tgcttctctt cgatcttcgc cagcagggcg
     2761 aggatcgtgg catcaccgaa ccgcgccgtg cgcgggtcgt cggtgagcca gagtttcagc
     2821 aggccgccca ggcggcccag gtcgccattg atgcgggcca gctcgcggac gtgctcatag
     2881 tccacgacgc ccgtgatttt gtagccctgg ccgacggcca gcaggtaggc cgacaggctc
     2941 atgccggccg ccgccgcctt ttcctcaatc gctcttcgtt cgtctggaag gcagtacacc
     3001 ttgataggtg ggctgccctt cctggttggc ttggtttcat cagccatccg cttgccctca
     3061 tctgttacgc cggcggtagc cggccagcct cgcagagcag gattcccgtt gagcaccgcc
     3121 aggtgcgaat aagggacagt gaagaaggaa cacccgctcg cgggtgggcc tacttcacct
     3181 atcctgcccg gctgacgccg ttggatacac caaggaaagt ctacacgaac cctttggcaa
     3241 aatcctgtat atcgtgcgaa aaaggatgga tataccgaaa aaatcgctat aatgaccccg
     3301 aagcagggtt atgcagcgga aaagcgctgc ttccctgctg ttttgtggaa tatctaccga
     3361 ctggaaacag gcaaatgcag gaaattactg aactgagggg acaggcgaga gacgatgcca
     3421 aagagctaca ccgacgagct ggccgagtgg gttgaatccc gcgcggccaa gaagcgccgg
     3481 cgtgatgagg ctgcggttgc gttcctggcg gtgagggcgg atgtcgaggc ggcgttagcg
     3541 tccggctatg cgctcgtcac catttgggag cacatgcggg aaacggggaa ggtcaagttc
     3601 tcctacgaga cgttccgctc gcacgccagg cggcacatca aggccaagcc cgccgatgtg
     3661 cccgcaccgc aggccaaggc tgcggaaccc gcgccggcac ccaagacgcc ggagccacgg
     3721 cggccgaagc aggggggcaa ggctgaaaag ccggcccccg ctgcggcccc gaccggcttc
     3781 accttcaacc caacaccgga caaaaaggat ccccaattct catgtttgac agcttatcat
     3841 cgataagctt taatgcggta gtttatcaca gttaaattgc taacgcagtc aggcaccgtg
     3901 tatgaaatct aacaatgcgc tcatcgtcat cctcggcacc gtcaccctgg atgctgtagg
     3961 cataggcttg gttatgccgg tactgccggg cctcttgcgg gatatcgtcc attccgacag
     4021 catcgccagt cactatggcg tgctgctagc gctatatgcg ttgatgcaat ttctatgcgc
     4081 acccgttctc ggagcactgt ccgaccgctt tggccgccgc ccagtcctgc tcgcttcgct
     4141 acttggagcc actatcgact acgcgatcat ggcgaccaca cccgtcctgt ggatcctcta
     4201 cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat
     4261 cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt
     4321 cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca
     4381 tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct
     4441 aatgcaggag tcgcataagg gagagcgtcg accgatgccc ttgagagcct tcaacccagt
     4501 cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt
     4561 tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg
     4621 ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc
     4681 cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat
     4741 tatcgccggc atggcggccg acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg
     4801 ctggatggcc ttccccatta tgattcttct cgcttccggc ggcatcggga tgcccgcgtt
     4861 gcaggccatg ctgtccaggc aggtagatga cgaccatcag ggacagcttc aaggatcgct
     4921 cgcggctctt accagcctaa cttcgatcac tggaccgctg atcgtcacgg cgatttatgc
     4981 cgcctcggcg agcacatgga acgggttggc atggattgta ggcgccgccc tataccttgt
     5041 ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct gaatggaagc
     5101 cggcggcacc tcgctaacgg attcaccact ccaagaattg gagccaatca attcttgcgg
     5161 agaactgtga atgcgcaaac caacccttgg cagaacatat ccatcgcgtc cgccatctcc
     5221 agcagccgca cgcggcgcat ctcgggcagc gttgggtcct ggccgggaat tacgccccgc
     5281 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
     5341 cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
     5401 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
     5461 caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc
     5521 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta
     5581 gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct
     5641 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
     5701 ttgccatacg gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg
     5761 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa
     5821 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat
     5881 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
     5941 tagctcctgc cctatgggat tcacctttat gttgataaga aataaaagaa aatgccaata
     6001 ggatatcggc attttctttt gcgtttttat ttgttaactg ttaattgtcc ttgttcaagg
     6061 atgctgtctt tgacaacaga tgttttcttg cctttgatgt tcagcaggaa gcttggcgca
     6121 aacgttgatt gtttgtctgc gtagaatcct ctgtttgtca tatagcttgt aatcacgaca
     6181 ttgtttcctt tcgcttgagg tacagcgaag tgtgagtaag taaaggttac atcgttagga
     6241 tcaagatcca tttttaacac aaggccagtt ttgttcagcg gcttgtatgg gccagttaaa
     6301 gaattagaaa cataaccaag catgtaaata tcgttagacg taatgccgtc aatcgtcatt
     6361 tttgatccgc gggagtcagt gaacaggtac catttgccgt tcattttaaa gacgttcgcg
     6421 cgttcaattt catctgttac tgtgttagat gcaatcagcg gtttcatcac ttttttcagt
     6481 gtgtaatcat cgtttagctc aatcataccg agagcgccgt ttgctaactc agccgtgcgt
     6541 tttttatcgc tttgcagaag tttttgactt tcttgacgga agaatgatgt gcttttgcca
     6601 tagtatgctt tgttaaataa agattcttcg ccttggtagc catcttcagt tccagtgttt
     6661 gcttcaaata ctaagtattt gtggccttta tcttctacgt agtgaggatc tctcagcgta
     6721 tggttgtcgc ctgagctgta gttgccttca tcgatgaact gctgtacatt ttgatacgtt
     6781 tttccgtcac cgtcaaagat tgatttataa tcctctacac cgttgatgtt caaagagctg
     6841 tctgatgctg atacgttaac ttgtgcagtt gtcagtgttt gtttgccgta atgtttaccg
     6901 gagaaatcag tgtagaataa acggattttt ccgtcagatg taaatgtggc tgaacctgac
     6961 cattcttgtg tttggtcttt taggatagaa tcatttgcat cgaatttgtc gctgtcttta
     7021 aagacgcggc cagcgttttt ccagctgtca atagaagttt cgccgacttt ttgatagaac
     7081 atgtaaatcg atgtgtcatc cgcattttta ggatctccgg ctaatgcaaa gacgatgtgg
     7141 tagccgtgat agtttgcgac agtgccgtca gcgttttgta atggccagct gtcccaaacg
     7201 tccaggcctt ttgcagaaga gatattttta attgtggacg aatcgaattc aggaacttga
     7261 tatttttcat ttttttgctg ttcagggatt tgcagcatat catggcgtgt aatatgggaa
     7321 atgccgtatg tttccttata tggcttttgg ttcgtttctt tcgcaaacgc ttgagttgcg
     7381 cctcctgcca gcagtgcggt agtaaaggtt aatactgttg cttgttttgc aaactttttg
     7441 atgttcatcg ttcatgtctc cttttttatg tactgtgtta gcggtctgct tcttccagcc
     7501 ctcctgtttg aagatggcaa gttagttacg cacaataaaa aaagacctaa aatatgtaag
     7561 gggtgacgcc aaagtataca ctttgccctt tacacatttt aggtcttgcc tgctttatca
     7621 gtaacaaacc cgcgcgattt acttttcgac ctcattctat tagactctcg tttggattgc
     7681 aactggtcta ttttcctctt ttgtttgata gaaaatcata aaaggatttg cagactacgg
     7741 gcctaaagaa ctaaaaaatc tatctgtttc ttttcattct ctgtattttt tatagtttct
     7801 gttgcatggg cataaagttg cctttttaat cacaattcag aaaatatcat aatatctcat
     7861 ttcactaaat aatagtgaac ggcaggtata tgtgatgggt taaaaaggat cgatcctcta
     7921 gccacgggtg cgcatgatcg tgctcctgtc gttgaggacc cggctaggct ggcggggttg
     7981 ccttactggt tagcagaatg aatcaccgat acgcgagcga acgtgaagcg actgctgctg
     8041 caaaacgtct gcgacctgag caacaacatg aatggtcttc ggtttccgtg tttcgtaaag
     8101 tctggaaacg cggaagtcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     8161 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     8221 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     8281 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     8341 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     8401 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     8461 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     8521 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     8581 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     8641 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     8701 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     8761 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     8821 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     8881 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     8941 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     9001 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     9061 acag
//
