LOCUS       40924_24193        8624 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHC38, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8624)
  AUTHORS   Chou H-H., Berthet J, Marx CJ.
  TITLE     Growth rate modulates the selective advantage of recurrent 
            transposition mutations arising from experimental populations grown 
            in a metal-limited environment
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8624)
  AUTHORS   Chou H-H.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-OCT-2008) Organismic and Evolutionary Biology, Harvard
            University, 16 Divinity Avenue, Biolab 3079, Cambridge, MA 02138, 
            USA
REFERENCE   3  (bases 1 to 8624)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8624)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-OCT-2008) Organismic and Evolutionary Biology, Harvard 
            University, 16 Divinity Avenue, Biolab 3079, Cambridge, MA 02138, 
            USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8624
                     /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    complement(1053..1631)
                     /label=gshA.CM1145 allele of Methylobacterium extorquens
                     /note="gshA.CM1145 allele of Methylobacterium extorquens"
     CDS             complement(2239..2607)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(2640..2749)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     promoter        3386..3414
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             3462..4649
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     CDS             complement(4833..5489)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             complement(5590..7008)
                     /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(7009..7454)
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     rep_origin      complement(7866..8454)
                     /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 tgacgtctag atctgaattc agctgtacaa
     1021 ttggtaccat ggatgcatat gctgcagctc gagatcgtct ccaccctgat cgcgcgactt
     1081 ccggtggtgc tcgcctcccg caccgagacc ggtcctgtgt tctcgcgcac atacgggtat
     1141 cccggcggtg agatcgatct gctgggtcgc ggcgcgctct cggccggcat cctgtcgggc
     1201 gtgaaagcgc ggctgatgct ccagcttctc cttcgggcgg gggtggatcg ggccgcctac
     1261 ggcgcctatt tcgcggatcg gtgatcggcg gggcagacgc cgcaaatatt agcttatgtc
     1321 atgtctgcca tgcggtcggc gggcgttgtt cactcttgca gcgtcctcta aggacacgac
     1381 atccggtgcg gctgctgacg gcccctccgg tggcggattc tcggcgcgcc gctgacccct
     1441 tcgaccgacg ttaccttcga ggcagcgcat ggcgcgcgac acgtccgaca ccaccccgct
     1501 cacgacgcgg gacgaactga tcgcgtggtt cgccgcgggc gagaaaccgc gcgcacggtt
     1561 cgcgatcggc accgagcacg agaagatccc gttctacacc gccgaccaca tgcccgttcc
     1621 atatgagggc gagctcacta gaggatccag ccgaccaggc tttccacgcc cgcgtgccgc
     1681 tccatgtcgt tcgcgcggtt ctcggaaacg cgctgccgcg tttcgtgatt gtcacgctca
     1741 agcccgtagt cccgttcgag cgtcgcgcag aggtcagcga gggcgcggta ggcccgatac
     1801 ggctcatgga tggtgtttcg ggtcgggtga atcttgttga tggcgatatg gatgtgcagg
     1861 ttgtcggtgt cgtgatgcac ggcactgacg cgctgatgct cggcgaagcc aagcccagcg
     1921 cagatgcggt cctcaatcgc gcgcaacgtc tccgcgtcgg gcttctctcc cgcgcggaag
     1981 ctaaccagca ggtgataggt cttgtcggcc tcggaacggg tgttgccgtg ctgggtcgcc
     2041 atcacctcgg ccatgacagc gggcagggtg tttgcctcgc agttcgtgac gcgcacgtga
     2101 cccaggcgct cggtcttgcc ttgctcgtcg gtgatgtact tcaccagctc cgcgaagtcg
     2161 ctcttcttga tggagcgcat ggggacgtgc ttggcaatca cgcgcacccc ccggccgttt
     2221 tagcggctaa aaaagtcatg gctctgccct cgggcggacc acgcccatca tgaccttgcc
     2281 aagctcgtcc tgcttctctt cgatcttcgc cagcagggcg aggatcgtgg catcaccgaa
     2341 ccgcgccgtg cgcgggtcgt cggtgagcca gagtttcagc aggccgccca ggcggcccag
     2401 gtcgccattg atgcgggcca gctcgcggac gtgctcatag tccacgacgc ccgtgatttt
     2461 gtagccctgg ccgacggcca gcaggtaggc cgacaggctc atgccggccg ccgccgcctt
     2521 ttcctcaatc gctcttcgtt cgtctggaag gcagtacacc ttgataggtg ggctgccctt
     2581 cctggttggc ttggtttcat cagccatccg cttgccctca tctgttacgc cggcggtagc
     2641 cggccagcct cgcagagcag gattcccgtt gagcaccgcc aggtgcgaat aagggacagt
     2701 gaagaaggaa cacccgctcg cgggtgggcc tacttcacct atcctgcccg gctgacgccg
     2761 ttggatacac caaggaaagt ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa
     2821 aaaggatgga tataccgaaa aaatcgctat aatgaccccg aagcagggtt atgcagcgga
     2881 aaagcgctgc ttccctgctg ttttgtggaa tatctaccga ctggaaacag gcaaatgcag
     2941 gaaattactg aactgagggg acaggcgaga gacgatgcca aagagctaca ccgacgagct
     3001 ggccgagtgg gttgaatccc gcgcggccaa gaagcgccgg cgtgatgagg ctgcggttgc
     3061 gttcctggcg gtgagggcgg atgtcgaggc ggcgttagcg tccggctatg cgctcgtcac
     3121 catttgggag cacatgcggg aaacggggaa ggtcaagttc tcctacgaga cgttccgctc
     3181 gcacgccagg cggcacatca aggccaagcc cgccgatgtg cccgcaccgc aggccaaggc
     3241 tgcggaaccc gcgccggcac ccaagacgcc ggagccacgg cggccgaagc aggggggcaa
     3301 ggctgaaaag ccggcccccg ctgcggcccc gaccggcttc accttcaacc caacaccgga
     3361 caaaaaggat ccccaattct catgtttgac agcttatcat cgataagctt taatgcggta
     3421 gtttatcaca gttaaattgc taacgcagtc aggcaccgtg tatgaaatct aacaatgcgc
     3481 tcatcgtcat cctcggcacc gtcaccctgg atgctgtagg cataggcttg gttatgccgg
     3541 tactgccggg cctcttgcgg gatatcgtcc attccgacag catcgccagt cactatggcg
     3601 tgctgctagc gctatatgcg ttgatgcaat ttctatgcgc acccgttctc ggagcactgt
     3661 ccgaccgctt tggccgccgc ccagtcctgc tcgcttcgct acttggagcc actatcgact
     3721 acgcgatcat ggcgaccaca cccgtcctgt ggatcctcta cgccggacgc atcgtggccg
     3781 gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg
     3841 aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt atggtggcag
     3901 gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc cttgcggcgg
     3961 cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag tcgcataagg
     4021 gagagcgtcg accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc
     4081 ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac
     4141 aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga
     4201 tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca
     4261 ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccg
     4321 acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc ttccccatta
     4381 tgattcttct cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc
     4441 aggtagatga cgaccatcag ggacagcttc aaggatcgct cgcggctctt accagcctaa
     4501 cttcgatcac tggaccgctg atcgtcacgg cgatttatgc cgcctcggcg agcacatgga
     4561 acgggttggc atggattgta ggcgccgccc tataccttgt ctgcctcccc gcgttgcgtc
     4621 gcggtgcatg gagccgggcc acctcgacct gaatggaagc cggcggcacc tcgctaacgg
     4681 attcaccact ccaagaattg gagccaatca attcttgcgg agaactgtga atgcgcaaac
     4741 caacccttgg cagaacatat ccatcgcgtc cgccatctcc agcagccgca cgcggcgcat
     4801 ctcgggcagc gttgggtcct ggccgggaat tacgccccgc cctgccactc atcgcagtac
     4861 tgttgtaatt cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc
     4921 tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa
     4981 acgggggcga agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc
     5041 cagggattgg ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg
     5101 ttttcaccgt aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg
     5161 tggtattcac tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa
     5221 gggtgaacac tatcccatat caccagctca ccgtctttca ttgccatacg gaattccgga
     5281 tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt
     5341 ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat
     5401 tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg
     5461 gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctgc cctatgggat
     5521 tcacctttat gttgataaga aataaaagaa aatgccaata ggatatcggc attttctttt
     5581 gcgtttttat ttgttaactg ttaattgtcc ttgttcaagg atgctgtctt tgacaacaga
     5641 tgttttcttg cctttgatgt tcagcaggaa gcttggcgca aacgttgatt gtttgtctgc
     5701 gtagaatcct ctgtttgtca tatagcttgt aatcacgaca ttgtttcctt tcgcttgagg
     5761 tacagcgaag tgtgagtaag taaaggttac atcgttagga tcaagatcca tttttaacac
     5821 aaggccagtt ttgttcagcg gcttgtatgg gccagttaaa gaattagaaa cataaccaag
     5881 catgtaaata tcgttagacg taatgccgtc aatcgtcatt tttgatccgc gggagtcagt
     5941 gaacaggtac catttgccgt tcattttaaa gacgttcgcg cgttcaattt catctgttac
     6001 tgtgttagat gcaatcagcg gtttcatcac ttttttcagt gtgtaatcat cgtttagctc
     6061 aatcataccg agagcgccgt ttgctaactc agccgtgcgt tttttatcgc tttgcagaag
     6121 tttttgactt tcttgacgga agaatgatgt gcttttgcca tagtatgctt tgttaaataa
     6181 agattcttcg ccttggtagc catcttcagt tccagtgttt gcttcaaata ctaagtattt
     6241 gtggccttta tcttctacgt agtgaggatc tctcagcgta tggttgtcgc ctgagctgta
     6301 gttgccttca tcgatgaact gctgtacatt ttgatacgtt tttccgtcac cgtcaaagat
     6361 tgatttataa tcctctacac cgttgatgtt caaagagctg tctgatgctg atacgttaac
     6421 ttgtgcagtt gtcagtgttt gtttgccgta atgtttaccg gagaaatcag tgtagaataa
     6481 acggattttt ccgtcagatg taaatgtggc tgaacctgac cattcttgtg tttggtcttt
     6541 taggatagaa tcatttgcat cgaatttgtc gctgtcttta aagacgcggc cagcgttttt
     6601 ccagctgtca atagaagttt cgccgacttt ttgatagaac atgtaaatcg atgtgtcatc
     6661 cgcattttta ggatctccgg ctaatgcaaa gacgatgtgg tagccgtgat agtttgcgac
     6721 agtgccgtca gcgttttgta atggccagct gtcccaaacg tccaggcctt ttgcagaaga
     6781 gatattttta attgtggacg aatcgaattc aggaacttga tatttttcat ttttttgctg
     6841 ttcagggatt tgcagcatat catggcgtgt aatatgggaa atgccgtatg tttccttata
     6901 tggcttttgg ttcgtttctt tcgcaaacgc ttgagttgcg cctcctgcca gcagtgcggt
     6961 agtaaaggtt aatactgttg cttgttttgc aaactttttg atgttcatcg ttcatgtctc
     7021 cttttttatg tactgtgtta gcggtctgct tcttccagcc ctcctgtttg aagatggcaa
     7081 gttagttacg cacaataaaa aaagacctaa aatatgtaag gggtgacgcc aaagtataca
     7141 ctttgccctt tacacatttt aggtcttgcc tgctttatca gtaacaaacc cgcgcgattt
     7201 acttttcgac ctcattctat tagactctcg tttggattgc aactggtcta ttttcctctt
     7261 ttgtttgata gaaaatcata aaaggatttg cagactacgg gcctaaagaa ctaaaaaatc
     7321 tatctgtttc ttttcattct ctgtattttt tatagtttct gttgcatggg cataaagttg
     7381 cctttttaat cacaattcag aaaatatcat aatatctcat ttcactaaat aatagtgaac
     7441 ggcaggtata tgtgatgggt taaaaaggat cgatcctcta gccacgggtg cgcatgatcg
     7501 tgctcctgtc gttgaggacc cggctaggct ggcggggttg ccttactggt tagcagaatg
     7561 aatcaccgat acgcgagcga acgtgaagcg actgctgctg caaaacgtct gcgacctgag
     7621 caacaacatg aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag
     7681 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     7741 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     7801 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     7861 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     7921 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     7981 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     8041 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     8101 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     8161 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     8221 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     8281 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     8341 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     8401 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     8461 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     8521 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     8581 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acag
//
