LOCUS       40924_24253        9157 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHC67, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9157)
  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  JOURNAL   Unpublished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 9157)
  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 9157)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9157)
  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            "Unpublished"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..9157
                     /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    1003..1593
                     /note="5' upstream region of Methylobacterium extorquens
                     AM1 icuB"
     misc_feature    1599..2163
                     /note="3' downstream region of Methylobacterium extorquens
                     AM1 icuB"
     CDS             complement(2772..3140)
                     /label=traJ
                     /note="oriT-recognizing protein"
     oriT            complement(3173..3282)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     promoter        3919..3947
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             3995..5182
                     /label=TcR
                     /note="tetracycline efflux protein"
     CDS             complement(5366..6022)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             complement(6123..7541)
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     promoter        complement(7542..7987)
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     rep_origin      complement(8399..8987)
                     /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 atctcggcac ggtgttcttc
     1021 accgacatcg ccaacgaggt cttccagccc gcacccggcc tgcccgtgca gaacctgggc
     1081 aagtcccgcc gcgacggctt cgaaatcgag gcgaaggtgc tggcctatcg tgaaggcctc
     1141 gaccgcgtgg cgttgttcgc caactattcg ggcgtctcgg cccgactgct caacggcgcg
     1201 ccatccatct tcatccccaa cgtgccggcg tatcttgcca ccgtcggcgc cgaattcgac
     1261 gtcgccgcgc cggccgccct gttgcccggc gggcgcctga ccggtgcggc ctacctgtcc
     1321 tacatcggtg ccaagccgtt gaccgaggac ggcgtattgc attcctcgcc ctatccgcgc
     1381 gtcagcggcc ggatcgccta cgtccaccag agcggctgga cgacctacgg gcaggccacc
     1441 tggtatcccg gcgaccgcac gagcgagagc atcttcaact tcggctcgtc ggtctctgcc
     1501 tcggccgccg agatcgtcac cagcccgcag ccgcgcctga cgctgctggc cggcatcacc
     1561 tccagctttg caacaggagc acctcttcga tgactcgagc gcgaggctcc cttgagatca
     1621 ggcggcatcg gctctcaaag ccttcgccgc ctcgatcaag gtccacgacg ccgaacggca
     1681 tcgcttgctc gcccagacag agcgaggcct tacgtcttca agactggtga gggctcacag
     1741 gccgcaacga gaggcgaggc gctttacggc gtccgccaag agagcggatg aactcatggc
     1801 gtcgcagcgc ggtgagcgtt catcgccctg aggcacgggg atgtggcggg tgcgttcgga
     1861 gagcaccgcc acgcgagcgg agttcgcctg aattcaggcg aactccgtcg atccgctgcg
     1921 ctcccggatt gatgctgccg tgctcgcatc acggggcggt tccaggcgag gactttcgga
     1981 tcggctgcaa aggaagaagc ggatgcaggc cgaagactcg gcatgccgtg ccgaggacga
     2041 gaccgcggcg acgcaggtgc gccgggtcga ccgcgacgaa caacacgcag gctggtgggc
     2101 ttctcgcacg tgcggtcgtt ccgggcggtc tgagggcggc cgcgatacgc tctcacggcc
     2161 tccgagctca ctagaggatc cagccgacca ggctttccac gcccgcgtgc cgctccatgt
     2221 cgttcgcgcg gttctcggaa acgcgctgcc gcgtttcgtg attgtcacgc tcaagcccgt
     2281 agtcccgttc gagcgtcgcg cagaggtcag cgagggcgcg gtaggcccga tacggctcat
     2341 ggatggtgtt tcgggtcggg tgaatcttgt tgatggcgat atggatgtgc aggttgtcgg
     2401 tgtcgtgatg cacggcactg acgcgctgat gctcggcgaa gccaagccca gcgcagatgc
     2461 ggtcctcaat cgcgcgcaac gtctccgcgt cgggcttctc tcccgcgcgg aagctaacca
     2521 gcaggtgata ggtcttgtcg gcctcggaac gggtgttgcc gtgctgggtc gccatcacct
     2581 cggccatgac agcgggcagg gtgtttgcct cgcagttcgt gacgcgcacg tgacccaggc
     2641 gctcggtctt gccttgctcg tcggtgatgt acttcaccag ctccgcgaag tcgctcttct
     2701 tgatggagcg catggggacg tgcttggcaa tcacgcgcac cccccggccg ttttagcggc
     2761 taaaaaagtc atggctctgc cctcgggcgg accacgccca tcatgacctt gccaagctcg
     2821 tcctgcttct cttcgatctt cgccagcagg gcgaggatcg tggcatcacc gaaccgcgcc
     2881 gtgcgcgggt cgtcggtgag ccagagtttc agcaggccgc ccaggcggcc caggtcgcca
     2941 ttgatgcggg ccagctcgcg gacgtgctca tagtccacga cgcccgtgat tttgtagccc
     3001 tggccgacgg ccagcaggta ggccgacagg ctcatgccgg ccgccgccgc cttttcctca
     3061 atcgctcttc gttcgtctgg aaggcagtac accttgatag gtgggctgcc cttcctggtt
     3121 ggcttggttt catcagccat ccgcttgccc tcatctgtta cgccggcggt agccggccag
     3181 cctcgcagag caggattccc gttgagcacc gccaggtgcg aataagggac agtgaagaag
     3241 gaacacccgc tcgcgggtgg gcctacttca cctatcctgc ccggctgacg ccgttggata
     3301 caccaaggaa agtctacacg aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat
     3361 ggatataccg aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc ggaaaagcgc
     3421 tgcttccctg ctgttttgtg gaatatctac cgactggaaa caggcaaatg caggaaatta
     3481 ctgaactgag gggacaggcg agagacgatg ccaaagagct acaccgacga gctggccgag
     3541 tgggttgaat cccgcgcggc caagaagcgc cggcgtgatg aggctgcggt tgcgttcctg
     3601 gcggtgaggg cggatgtcga ggcggcgtta gcgtccggct atgcgctcgt caccatttgg
     3661 gagcacatgc gggaaacggg gaaggtcaag ttctcctacg agacgttccg ctcgcacgcc
     3721 aggcggcaca tcaaggccaa gcccgccgat gtgcccgcac cgcaggccaa ggctgcggaa
     3781 cccgcgccgg cacccaagac gccggagcca cggcggccga agcagggggg caaggctgaa
     3841 aagccggccc ccgctgcggc cccgaccggc ttcaccttca acccaacacc ggacaaaaag
     3901 gatccccaat tctcatgttt gacagcttat catcgataag ctttaatgcg gtagtttatc
     3961 acagttaaat tgctaacgca gtcaggcacc gtgtatgaaa tctaacaatg cgctcatcgt
     4021 catcctcggc accgtcaccc tggatgctgt aggcataggc ttggttatgc cggtactgcc
     4081 gggcctcttg cgggatatcg tccattccga cagcatcgcc agtcactatg gcgtgctgct
     4141 agcgctatat gcgttgatgc aatttctatg cgcacccgtt ctcggagcac tgtccgaccg
     4201 ctttggccgc cgcccagtcc tgctcgcttc gctacttgga gccactatcg actacgcgat
     4261 catggcgacc acacccgtcc tgtggatcct ctacgccgga cgcatcgtgg ccggcatcac
     4321 cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg
     4381 ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt
     4441 ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct
     4501 caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg
     4561 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
     4621 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
     4681 ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg
     4741 cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc
     4801 cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct
     4861 gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca ttatgattct
     4921 tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga
     4981 tgacgaccat cagggacagc ttcaaggatc gctcgcggct cttaccagcc taacttcgat
     5041 cactggaccg ctgatcgtca cggcgattta tgccgcctcg gcgagcacat ggaacgggtt
     5101 ggcatggatt gtaggcgccg ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc
     5161 atggagccgg gccacctcga cctgaatgga agccggcggc acctcgctaa cggattcacc
     5221 actccaagaa ttggagccaa tcaattcttg cggagaactg tgaatgcgca aaccaaccct
     5281 tggcagaaca tatccatcgc gtccgccatc tccagcagcc gcacgcggcg catctcgggc
     5341 agcgttgggt cctggccggg aattacgccc cgccctgcca ctcatcgcag tactgttgta
     5401 attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg
     5461 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg
     5521 cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat
     5581 tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac
     5641 cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
     5701 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
     5761 cactatccca tatcaccagc tcaccgtctt tcattgccat acggaattcc ggatgagcat
     5821 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
     5881 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     5941 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     6001 atccagtgat ttttttctcc attttagctt ccttagctcc tgccctatgg gattcacctt
     6061 tatgttgata agaaataaaa gaaaatgcca ataggatatc ggcattttct tttgcgtttt
     6121 tatttgttaa ctgttaattg tccttgttca aggatgctgt ctttgacaac agatgttttc
     6181 ttgcctttga tgttcagcag gaagcttggc gcaaacgttg attgtttgtc tgcgtagaat
     6241 cctctgtttg tcatatagct tgtaatcacg acattgtttc ctttcgcttg aggtacagcg
     6301 aagtgtgagt aagtaaaggt tacatcgtta ggatcaagat ccatttttaa cacaaggcca
     6361 gttttgttca gcggcttgta tgggccagtt aaagaattag aaacataacc aagcatgtaa
     6421 atatcgttag acgtaatgcc gtcaatcgtc atttttgatc cgcgggagtc agtgaacagg
     6481 taccatttgc cgttcatttt aaagacgttc gcgcgttcaa tttcatctgt tactgtgtta
     6541 gatgcaatca gcggtttcat cacttttttc agtgtgtaat catcgtttag ctcaatcata
     6601 ccgagagcgc cgtttgctaa ctcagccgtg cgttttttat cgctttgcag aagtttttga
     6661 ctttcttgac ggaagaatga tgtgcttttg ccatagtatg ctttgttaaa taaagattct
     6721 tcgccttggt agccatcttc agttccagtg tttgcttcaa atactaagta tttgtggcct
     6781 ttatcttcta cgtagtgagg atctctcagc gtatggttgt cgcctgagct gtagttgcct
     6841 tcatcgatga actgctgtac attttgatac gtttttccgt caccgtcaaa gattgattta
     6901 taatcctcta caccgttgat gttcaaagag ctgtctgatg ctgatacgtt aacttgtgca
     6961 gttgtcagtg tttgtttgcc gtaatgttta ccggagaaat cagtgtagaa taaacggatt
     7021 tttccgtcag atgtaaatgt ggctgaacct gaccattctt gtgtttggtc ttttaggata
     7081 gaatcatttg catcgaattt gtcgctgtct ttaaagacgc ggccagcgtt tttccagctg
     7141 tcaatagaag tttcgccgac tttttgatag aacatgtaaa tcgatgtgtc atccgcattt
     7201 ttaggatctc cggctaatgc aaagacgatg tggtagccgt gatagtttgc gacagtgccg
     7261 tcagcgtttt gtaatggcca gctgtcccaa acgtccaggc cttttgcaga agagatattt
     7321 ttaattgtgg acgaatcgaa ttcaggaact tgatattttt catttttttg ctgttcaggg
     7381 atttgcagca tatcatggcg tgtaatatgg gaaatgccgt atgtttcctt atatggcttt
     7441 tggttcgttt ctttcgcaaa cgcttgagtt gcgcctcctg ccagcagtgc ggtagtaaag
     7501 gttaatactg ttgcttgttt tgcaaacttt ttgatgttca tcgttcatgt ctcctttttt
     7561 atgtactgtg ttagcggtct gcttcttcca gccctcctgt ttgaagatgg caagttagtt
     7621 acgcacaata aaaaaagacc taaaatatgt aaggggtgac gccaaagtat acactttgcc
     7681 ctttacacat tttaggtctt gcctgcttta tcagtaacaa acccgcgcga tttacttttc
     7741 gacctcattc tattagactc tcgtttggat tgcaactggt ctattttcct cttttgtttg
     7801 atagaaaatc ataaaaggat ttgcagacta cgggcctaaa gaactaaaaa atctatctgt
     7861 ttcttttcat tctctgtatt ttttatagtt tctgttgcat gggcataaag ttgccttttt
     7921 aatcacaatt cagaaaatat cataatatct catttcacta aataatagtg aacggcaggt
     7981 atatgtgatg ggttaaaaag gatcgatcct ctagccacgg gtgcgcatga tcgtgctcct
     8041 gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga atgaatcacc
     8101 gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac
     8161 atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt cagcgctctt
     8221 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     8281 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     8341 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     8401 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     8461 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     8521 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     8581 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     8641 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     8701 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     8761 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     8821 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     8881 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     8941 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     9001 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     9061 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     9121 caatctaaag tatatatgag taaacttggt ctgacag
//
