LOCUS       40924_29306        9063 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLW17, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9063)
  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 9063)
  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 9063)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9063)
  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..9063
                     /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(2678..3046)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(3079..3188)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     promoter        3825..3853
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             3901..5088
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     CDS             complement(5272..5928)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             complement(6029..7447)
                     /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(7448..7893)
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     rep_origin      complement(8305..8893)
                     /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 ggtttgagct tcggccccgt ccagatgtcg ctcgcggcgg gctacgtgcg ggaccgcgtc
     1201 cagggcacgg gctactattc cagcatcgag gcgcgcgcga acttttagag cgcattccga
     1261 cggcgagcag gccgccgctc gcggcggcgg ccgaagtcaa gcgcttggat caatgcggac
     1321 aatccgccgc cttgtctccg cgtctcacgg caaagtactg ctgttccaag cgaagaatgg
     1381 ccctcatatc aggaagacat ccctcgccat cgcatgtcat ttccgcgaaa ttcctgccga
     1441 atcttcggac attttctacg gatgacgcta cgctctgcat cgcaaaatca accttgttcg
     1501 cagagcggga cggcggcgtc gtgaacgcat ccggaaatcg agatctcata tgcacgaagt
     1561 gaggacgcct tcgatctcta caagtcggtc ctcgatggtc ctgccgatgc gggcgcgcgg
     1621 ctcgccacga tccagaaggc gatgagccat ctcaagatgg atcaggccga gcggctcatc
     1681 gccatggggc gctccaaccc ggagggcgct tcggaattcg acgcgatccg cctcgacatc
     1741 acccgcgccc gcatcgccgc cttcctgcac gacgagccgg cccagacgcc gaccctggcc
     1801 gacctcaccg ccttccagac ctatgcccgc aagagcggag acccgagcca gaccggcctg
     1861 ctcggctggt acgcctacaa gcggcgccag ttccgcgagg cgctggaatg gttcaagctc
     1921 gccatcggcc gcggcggcga cgccatggtc gcccacggcc tcgcccacac cctgcgcgaa
     1981 ctgaacatgc tgcgcgaggc ggaagaggtc gcctatgcct ggcgcgaccg cttcgtcggc
     2041 aacgagctgc tgttcatcga cgtgctcgag agctcactag aggatccagc cgaccaggct
     2101 ttccacgccc gcgtgccgct ccatgtcgtt cgcgcggttc tcggaaacgc gctgccgcgt
     2161 ttcgtgattg tcacgctcaa gcccgtagtc ccgttcgagc gtcgcgcaga ggtcagcgag
     2221 ggcgcggtag gcccgatacg gctcatggat ggtgtttcgg gtcgggtgaa tcttgttgat
     2281 ggcgatatgg atgtgcaggt tgtcggtgtc gtgatgcacg gcactgacgc gctgatgctc
     2341 ggcgaagcca agcccagcgc agatgcggtc ctcaatcgcg cgcaacgtct ccgcgtcggg
     2401 cttctctccc gcgcggaagc taaccagcag gtgataggtc ttgtcggcct cggaacgggt
     2461 gttgccgtgc tgggtcgcca tcacctcggc catgacagcg ggcagggtgt ttgcctcgca
     2521 gttcgtgacg cgcacgtgac ccaggcgctc ggtcttgcct tgctcgtcgg tgatgtactt
     2581 caccagctcc gcgaagtcgc tcttcttgat ggagcgcatg gggacgtgct tggcaatcac
     2641 gcgcaccccc cggccgtttt agcggctaaa aaagtcatgg ctctgccctc gggcggacca
     2701 cgcccatcat gaccttgcca agctcgtcct gcttctcttc gatcttcgcc agcagggcga
     2761 ggatcgtggc atcaccgaac cgcgccgtgc gcgggtcgtc ggtgagccag agtttcagca
     2821 ggccgcccag gcggcccagg tcgccattga tgcgggccag ctcgcggacg tgctcatagt
     2881 ccacgacgcc cgtgattttg tagccctggc cgacggccag caggtaggcc gacaggctca
     2941 tgccggccgc cgccgccttt tcctcaatcg ctcttcgttc gtctggaagg cagtacacct
     3001 tgataggtgg gctgcccttc ctggttggct tggtttcatc agccatccgc ttgccctcat
     3061 ctgttacgcc ggcggtagcc ggccagcctc gcagagcagg attcccgttg agcaccgcca
     3121 ggtgcgaata agggacagtg aagaaggaac acccgctcgc gggtgggcct acttcaccta
     3181 tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc tacacgaacc ctttggcaaa
     3241 atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata atgaccccga
     3301 agcagggtta tgcagcggaa aagcgctgct tccctgctgt tttgtggaat atctaccgac
     3361 tggaaacagg caaatgcagg aaattactga actgagggga caggcgagag acgatgccaa
     3421 agagctacac cgacgagctg gccgagtggg ttgaatcccg cgcggccaag aagcgccggc
     3481 gtgatgaggc tgcggttgcg ttcctggcgg tgagggcgga tgtcgaggcg gcgttagcgt
     3541 ccggctatgc gctcgtcacc atttgggagc acatgcggga aacggggaag gtcaagttct
     3601 cctacgagac gttccgctcg cacgccaggc ggcacatcaa ggccaagccc gccgatgtgc
     3661 ccgcaccgca ggccaaggct gcggaacccg cgccggcacc caagacgccg gagccacggc
     3721 ggccgaagca ggggggcaag gctgaaaagc cggcccccgc tgcggccccg accggcttca
     3781 ccttcaaccc aacaccggac aaaaaggatc cccaattctc atgtttgaca gcttatcatc
     3841 gataagcttt aatgcggtag tttatcacag ttaaattgct aacgcagtca ggcaccgtgt
     3901 atgaaatcta acaatgcgct catcgtcatc ctcggcaccg tcaccctgga tgctgtaggc
     3961 ataggcttgg ttatgccggt actgccgggc ctcttgcggg atatcgtcca ttccgacagc
     4021 atcgccagtc actatggcgt gctgctagcg ctatatgcgt tgatgcaatt tctatgcgca
     4081 cccgttctcg gagcactgtc cgaccgcttt ggccgccgcc cagtcctgct cgcttcgcta
     4141 cttggagcca ctatcgacta cgcgatcatg gcgaccacac ccgtcctgtg gatcctctac
     4201 gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc
     4261 gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc
     4321 ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat
     4381 gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta
     4441 atgcaggagt cgcataaggg agagcgtcga ccgatgccct tgagagcctt caacccagtc
     4501 agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt
     4561 atcatgcaac tcgtaggaca ggtgccggca gcgctctggg tcattttcgg cgaggaccgc
     4621 tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc
     4681 ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt
     4741 atcgccggca tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc
     4801 tggatggcct tccccattat gattcttctc gcttccggcg gcatcgggat gcccgcgttg
     4861 caggccatgc tgtccaggca ggtagatgac gaccatcagg gacagcttca aggatcgctc
     4921 gcggctctta ccagcctaac ttcgatcact ggaccgctga tcgtcacggc gatttatgcc
     4981 gcctcggcga gcacatggaa cgggttggca tggattgtag gcgccgccct ataccttgtc
     5041 tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca cctcgacctg aatggaagcc
     5101 ggcggcacct cgctaacgga ttcaccactc caagaattgg agccaatcaa ttcttgcgga
     5161 gaactgtgaa tgcgcaaacc aacccttggc agaacatatc catcgcgtcc gccatctcca
     5221 gcagccgcac gcggcgcatc tcgggcagcg ttgggtcctg gccgggaatt acgccccgcc
     5281 ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc
     5341 acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
     5401 atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
     5461 aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc
     5521 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
     5581 aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc
     5641 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
     5701 tgccatacgg aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
     5761 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
     5821 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
     5881 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
     5941 agctcctgcc ctatgggatt cacctttatg ttgataagaa ataaaagaaa atgccaatag
     6001 gatatcggca ttttcttttg cgtttttatt tgttaactgt taattgtcct tgttcaagga
     6061 tgctgtcttt gacaacagat gttttcttgc ctttgatgtt cagcaggaag cttggcgcaa
     6121 acgttgattg tttgtctgcg tagaatcctc tgtttgtcat atagcttgta atcacgacat
     6181 tgtttccttt cgcttgaggt acagcgaagt gtgagtaagt aaaggttaca tcgttaggat
     6241 caagatccat ttttaacaca aggccagttt tgttcagcgg cttgtatggg ccagttaaag
     6301 aattagaaac ataaccaagc atgtaaatat cgttagacgt aatgccgtca atcgtcattt
     6361 ttgatccgcg ggagtcagtg aacaggtacc atttgccgtt cattttaaag acgttcgcgc
     6421 gttcaatttc atctgttact gtgttagatg caatcagcgg tttcatcact tttttcagtg
     6481 tgtaatcatc gtttagctca atcataccga gagcgccgtt tgctaactca gccgtgcgtt
     6541 ttttatcgct ttgcagaagt ttttgacttt cttgacggaa gaatgatgtg cttttgccat
     6601 agtatgcttt gttaaataaa gattcttcgc cttggtagcc atcttcagtt ccagtgtttg
     6661 cttcaaatac taagtatttg tggcctttat cttctacgta gtgaggatct ctcagcgtat
     6721 ggttgtcgcc tgagctgtag ttgccttcat cgatgaactg ctgtacattt tgatacgttt
     6781 ttccgtcacc gtcaaagatt gatttataat cctctacacc gttgatgttc aaagagctgt
     6841 ctgatgctga tacgttaact tgtgcagttg tcagtgtttg tttgccgtaa tgtttaccgg
     6901 agaaatcagt gtagaataaa cggatttttc cgtcagatgt aaatgtggct gaacctgacc
     6961 attcttgtgt ttggtctttt aggatagaat catttgcatc gaatttgtcg ctgtctttaa
     7021 agacgcggcc agcgtttttc cagctgtcaa tagaagtttc gccgactttt tgatagaaca
     7081 tgtaaatcga tgtgtcatcc gcatttttag gatctccggc taatgcaaag acgatgtggt
     7141 agccgtgata gtttgcgaca gtgccgtcag cgttttgtaa tggccagctg tcccaaacgt
     7201 ccaggccttt tgcagaagag atatttttaa ttgtggacga atcgaattca ggaacttgat
     7261 atttttcatt tttttgctgt tcagggattt gcagcatatc atggcgtgta atatgggaaa
     7321 tgccgtatgt ttccttatat ggcttttggt tcgtttcttt cgcaaacgct tgagttgcgc
     7381 ctcctgccag cagtgcggta gtaaaggtta atactgttgc ttgttttgca aactttttga
     7441 tgttcatcgt tcatgtctcc ttttttatgt actgtgttag cggtctgctt cttccagccc
     7501 tcctgtttga agatggcaag ttagttacgc acaataaaaa aagacctaaa atatgtaagg
     7561 ggtgacgcca aagtatacac tttgcccttt acacatttta ggtcttgcct gctttatcag
     7621 taacaaaccc gcgcgattta cttttcgacc tcattctatt agactctcgt ttggattgca
     7681 actggtctat tttcctcttt tgtttgatag aaaatcataa aaggatttgc agactacggg
     7741 cctaaagaac taaaaaatct atctgtttct tttcattctc tgtatttttt atagtttctg
     7801 ttgcatgggc ataaagttgc ctttttaatc acaattcaga aaatatcata atatctcatt
     7861 tcactaaata atagtgaacg gcaggtatat gtgatgggtt aaaaaggatc gatcctctag
     7921 ccacgggtgc gcatgatcgt gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc
     7981 cttactggtt agcagaatga atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc
     8041 aaaacgtctg cgacctgagc aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt
     8101 ctggaaacgc ggaagtcagc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     8161 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     8221 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     8281 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     8341 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     8401 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     8461 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     8521 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     8581 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     8641 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     8701 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     8761 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     8821 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     8881 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     8941 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     9001 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     9061 cag
//
