LOCUS       40924_24258        9191 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHC68, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9191)
  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  JOURNAL   Unpublished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 9191)
  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 9191)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9191)
  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            "Unpublished"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..9191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(4..861)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(862..966)
                     /label=AmpR promoter
     misc_feature    1005..1628
                     /note="5' upstream region of Methylobacterium extorquens
                     AM1 icuA"
     misc_feature    1633..2197
                     /note="3' downstream region of Methylobacterium extorquens
                     AM1 icuB"
     CDS             complement(2806..3174)
                     /label=traJ
                     /note="oriT-recognizing protein"
     oriT            complement(3207..3316)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     promoter        3953..3981
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             4029..5216
                     /label=TcR
                     /note="tetracycline efflux protein"
     CDS             complement(5400..6056)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             complement(6157..7575)
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     promoter        complement(7576..8021)
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     rep_origin      complement(8433..9021)
                     /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 atctgcgaat acagcctgcc
     1021 ttagcagccg atttattggc acccagaaca tatcttgttt tcaacctata gcagcaactt
     1081 aacgaacgtg gctagcccag tttgaatacg gggtattact tatccaaatg atcagtttcg
     1141 cgcataagtt gttgcaataa agggagattc gcgctgggct tggcggcccc tcacacttac
     1201 ggcctcggcg ccgaacggtt tccccatttc ggagcgggag gggcgccgag ttgcgacgca
     1261 agagttgcaa ggatcgctcc gccccgcatt ggaatgctcg attttagggc tgctgctccg
     1321 gctgcgatgg cctccggcca tcgtacgatg tgaagctcag aagccactgg cctcgaagac
     1381 gtttcgcgcc tgatggaccg aagacttggt ggccggcagg acatgccacg ccgtatcagc
     1441 accgatgctc taagggaggc aattttaccg agacggttat cgtcaaaaga tgctgcgacc
     1501 gagattgcgg gggctgctaa aggctgcaac atctcgatga gacgtacgaa tacgaaaaat
     1561 tcctatcttg tgcctgccca taatttgcag tttagtagcg cccggatcac tgagtgccta
     1621 ggttgcactc gagcgcgagg ctcccttgag atcaggcggc atcggctctc aaagccttcg
     1681 ccgcctcgat caaggtccac gacgccgaac ggcatcgctt gctcgcccag acagagcgag
     1741 gccttacgtc ttcaagactg gtgagggctc acaggccgca acgagaggcg aggcgcttta
     1801 cggcgtccgc caagagagcg gatgaactca tggcgtcgca gcgcggtgag cgttcatcgc
     1861 cctgaggcac ggggatgtgg cgggtgcgtt cggagagcac cgccacgcga gcggagttcg
     1921 cctgaattca ggcgaactcc gtcgatccgc tgcgctcccg gattgatgct gccgtgctcg
     1981 catcacgggg cggttccagg cgaggacttt cggatcggct gcaaaggaag aagcggatgc
     2041 aggccgaaga ctcggcatgc cgtgccgagg acgagaccgc ggcgacgcag gtgcgccggg
     2101 tcgaccgcga cgaacaacac gcaggctggt gggcttctcg cacgtgcggt cgttccgggc
     2161 ggtctgaggg cggccgcgat acgctctcac ggcctccgag ctcactagag gatccagccg
     2221 accaggcttt ccacgcccgc gtgccgctcc atgtcgttcg cgcggttctc ggaaacgcgc
     2281 tgccgcgttt cgtgattgtc acgctcaagc ccgtagtccc gttcgagcgt cgcgcagagg
     2341 tcagcgaggg cgcggtaggc ccgatacggc tcatggatgg tgtttcgggt cgggtgaatc
     2401 ttgttgatgg cgatatggat gtgcaggttg tcggtgtcgt gatgcacggc actgacgcgc
     2461 tgatgctcgg cgaagccaag cccagcgcag atgcggtcct caatcgcgcg caacgtctcc
     2521 gcgtcgggct tctctcccgc gcggaagcta accagcaggt gataggtctt gtcggcctcg
     2581 gaacgggtgt tgccgtgctg ggtcgccatc acctcggcca tgacagcggg cagggtgttt
     2641 gcctcgcagt tcgtgacgcg cacgtgaccc aggcgctcgg tcttgccttg ctcgtcggtg
     2701 atgtacttca ccagctccgc gaagtcgctc ttcttgatgg agcgcatggg gacgtgcttg
     2761 gcaatcacgc gcaccccccg gccgttttag cggctaaaaa agtcatggct ctgccctcgg
     2821 gcggaccacg cccatcatga ccttgccaag ctcgtcctgc ttctcttcga tcttcgccag
     2881 cagggcgagg atcgtggcat caccgaaccg cgccgtgcgc gggtcgtcgg tgagccagag
     2941 tttcagcagg ccgcccaggc ggcccaggtc gccattgatg cgggccagct cgcggacgtg
     3001 ctcatagtcc acgacgcccg tgattttgta gccctggccg acggccagca ggtaggccga
     3061 caggctcatg ccggccgccg ccgccttttc ctcaatcgct cttcgttcgt ctggaaggca
     3121 gtacaccttg ataggtgggc tgcccttcct ggttggcttg gtttcatcag ccatccgctt
     3181 gccctcatct gttacgccgg cggtagccgg ccagcctcgc agagcaggat tcccgttgag
     3241 caccgccagg tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac
     3301 ttcacctatc ctgcccggct gacgccgttg gatacaccaa ggaaagtcta cacgaaccct
     3361 ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat
     3421 gaccccgaag cagggttatg cagcggaaaa gcgctgcttc cctgctgttt tgtggaatat
     3481 ctaccgactg gaaacaggca aatgcaggaa attactgaac tgaggggaca ggcgagagac
     3541 gatgccaaag agctacaccg acgagctggc cgagtgggtt gaatcccgcg cggccaagaa
     3601 gcgccggcgt gatgaggctg cggttgcgtt cctggcggtg agggcggatg tcgaggcggc
     3661 gttagcgtcc ggctatgcgc tcgtcaccat ttgggagcac atgcgggaaa cggggaaggt
     3721 caagttctcc tacgagacgt tccgctcgca cgccaggcgg cacatcaagg ccaagcccgc
     3781 cgatgtgccc gcaccgcagg ccaaggctgc ggaacccgcg ccggcaccca agacgccgga
     3841 gccacggcgg ccgaagcagg ggggcaaggc tgaaaagccg gcccccgctg cggccccgac
     3901 cggcttcacc ttcaacccaa caccggacaa aaaggatccc caattctcat gtttgacagc
     3961 ttatcatcga taagctttaa tgcggtagtt tatcacagtt aaattgctaa cgcagtcagg
     4021 caccgtgtat gaaatctaac aatgcgctca tcgtcatcct cggcaccgtc accctggatg
     4081 ctgtaggcat aggcttggtt atgccggtac tgccgggcct cttgcgggat atcgtccatt
     4141 ccgacagcat cgccagtcac tatggcgtgc tgctagcgct atatgcgttg atgcaatttc
     4201 tatgcgcacc cgttctcgga gcactgtccg accgctttgg ccgccgccca gtcctgctcg
     4261 cttcgctact tggagccact atcgactacg cgatcatggc gaccacaccc gtcctgtgga
     4321 tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg
     4381 cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg
     4441 cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct
     4501 ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct
     4561 gcttcctaat gcaggagtcg cataagggag agcgtcgacc gatgcccttg agagccttca
     4621 acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg
     4681 tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg
     4741 aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct
     4801 tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc
     4861 aggccattat cgccggcatg gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga
     4921 cgcgaggctg gatggccttc cccattatga ttcttctcgc ttccggcggc atcgggatgc
     4981 ccgcgttgca ggccatgctg tccaggcagg tagatgacga ccatcaggga cagcttcaag
     5041 gatcgctcgc ggctcttacc agcctaactt cgatcactgg accgctgatc gtcacggcga
     5101 tttatgccgc ctcggcgagc acatggaacg ggttggcatg gattgtaggc gccgccctat
     5161 accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa
     5221 tggaagccgg cggcacctcg ctaacggatt caccactcca agaattggag ccaatcaatt
     5281 cttgcggaga actgtgaatg cgcaaaccaa cccttggcag aacatatcca tcgcgtccgc
     5341 catctccagc agccgcacgc ggcgcatctc gggcagcgtt gggtcctggc cgggaattac
     5401 gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg
     5461 aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct
     5521 tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg
     5581 tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca
     5641 ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat
     5701 atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca
     5761 gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg
     5821 tctttcattg ccatacggaa ttccggatga gcattcatca ggcgggcaag aatgtgaata
     5881 aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc
     5941 agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct
     6001 ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta
     6061 gcttccttag ctcctgccct atgggattca cctttatgtt gataagaaat aaaagaaaat
     6121 gccaatagga tatcggcatt ttcttttgcg tttttatttg ttaactgtta attgtccttg
     6181 ttcaaggatg ctgtctttga caacagatgt tttcttgcct ttgatgttca gcaggaagct
     6241 tggcgcaaac gttgattgtt tgtctgcgta gaatcctctg tttgtcatat agcttgtaat
     6301 cacgacattg tttcctttcg cttgaggtac agcgaagtgt gagtaagtaa aggttacatc
     6361 gttaggatca agatccattt ttaacacaag gccagttttg ttcagcggct tgtatgggcc
     6421 agttaaagaa ttagaaacat aaccaagcat gtaaatatcg ttagacgtaa tgccgtcaat
     6481 cgtcattttt gatccgcggg agtcagtgaa caggtaccat ttgccgttca ttttaaagac
     6541 gttcgcgcgt tcaatttcat ctgttactgt gttagatgca atcagcggtt tcatcacttt
     6601 tttcagtgtg taatcatcgt ttagctcaat cataccgaga gcgccgtttg ctaactcagc
     6661 cgtgcgtttt ttatcgcttt gcagaagttt ttgactttct tgacggaaga atgatgtgct
     6721 tttgccatag tatgctttgt taaataaaga ttcttcgcct tggtagccat cttcagttcc
     6781 agtgtttgct tcaaatacta agtatttgtg gcctttatct tctacgtagt gaggatctct
     6841 cagcgtatgg ttgtcgcctg agctgtagtt gccttcatcg atgaactgct gtacattttg
     6901 atacgttttt ccgtcaccgt caaagattga tttataatcc tctacaccgt tgatgttcaa
     6961 agagctgtct gatgctgata cgttaacttg tgcagttgtc agtgtttgtt tgccgtaatg
     7021 tttaccggag aaatcagtgt agaataaacg gatttttccg tcagatgtaa atgtggctga
     7081 acctgaccat tcttgtgttt ggtcttttag gatagaatca tttgcatcga atttgtcgct
     7141 gtctttaaag acgcggccag cgtttttcca gctgtcaata gaagtttcgc cgactttttg
     7201 atagaacatg taaatcgatg tgtcatccgc atttttagga tctccggcta atgcaaagac
     7261 gatgtggtag ccgtgatagt ttgcgacagt gccgtcagcg ttttgtaatg gccagctgtc
     7321 ccaaacgtcc aggccttttg cagaagagat atttttaatt gtggacgaat cgaattcagg
     7381 aacttgatat ttttcatttt tttgctgttc agggatttgc agcatatcat ggcgtgtaat
     7441 atgggaaatg ccgtatgttt ccttatatgg cttttggttc gtttctttcg caaacgcttg
     7501 agttgcgcct cctgccagca gtgcggtagt aaaggttaat actgttgctt gttttgcaaa
     7561 ctttttgatg ttcatcgttc atgtctcctt ttttatgtac tgtgttagcg gtctgcttct
     7621 tccagccctc ctgtttgaag atggcaagtt agttacgcac aataaaaaaa gacctaaaat
     7681 atgtaagggg tgacgccaaa gtatacactt tgccctttac acattttagg tcttgcctgc
     7741 tttatcagta acaaacccgc gcgatttact tttcgacctc attctattag actctcgttt
     7801 ggattgcaac tggtctattt tcctcttttg tttgatagaa aatcataaaa ggatttgcag
     7861 actacgggcc taaagaacta aaaaatctat ctgtttcttt tcattctctg tattttttat
     7921 agtttctgtt gcatgggcat aaagttgcct ttttaatcac aattcagaaa atatcataat
     7981 atctcatttc actaaataat agtgaacggc aggtatatgt gatgggttaa aaaggatcga
     8041 tcctctagcc acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc
     8101 ggggttgcct tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact
     8161 gctgctgcaa aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt
     8221 cgtaaagtct ggaaacgcgg aagtcagcgc tcttccgctt cctcgctcac tgactcgctg
     8281 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     8341 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     8401 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     8461 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     8521 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     8581 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     8641 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     8701 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     8761 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     8821 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     8881 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     8941 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     9001 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     9061 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     9121 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     9181 tggtctgaca g
//
