LOCUS       40924_24248        9177 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHC65, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9177)
  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  JOURNAL   Unpublished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 9177)
  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 9177)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9177)
  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            "Unpublished"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..9177
                     /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    1634..2182
                     /note="3' downstream region of Methylobacterium extorquens
                     AM1 icuA"
     CDS             complement(2792..3160)
                     /label=traJ
                     /note="oriT-recognizing protein"
     oriT            complement(3193..3302)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     promoter        3939..3967
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             4015..5202
                     /label=TcR
                     /note="tetracycline efflux protein"
     CDS             complement(5386..6042)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             complement(6143..7561)
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     promoter        complement(7562..8007)
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     rep_origin      complement(8419..9007)
                     /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 gagatgagac gaaccgtcat cctcaccgcg ctgctcgcct tcaccgcacc
     1681 ggccgccgcc cacgacctct ggctcgaccc ggccggcaac ggcgtgcaga tcctctacgg
     1741 gcatccccac gagccggaac tgccgagtgc tggcaagctg atgagcctga ccgcctacga
     1801 gccgtccggc gcggtggccc tgaatgccaa gctgcagacc ggcgcaatgc ccgcgctcaa
     1861 agccgcgcat cagggcgacg cgctgttcgc cgcggcctac gacaacggct actgggtgcg
     1921 cttgtccgac ggcggctacc gcaacgccag caagcggatg ctgccgcagg ccgacaagag
     1981 cctctggtcg gtgaagttcg ccaaggccgt atccggcccg acggccccgt gggacaagat
     2041 cgtcggccaa ccgctggaga tcgtcccgct ggaggagccg gccgcggctt cgggccagat
     2101 ccgggtgcgg gtgctgttcg agggccgtcc gctcgacggc gccagcgtgg tcgccaccga
     2161 cggcgtgaac ttcaagagcg agcgagctca ctagaggatc cagccgacca ggctttccac
     2221 gcccgcgtgc cgctccatgt cgttcgcgcg gttctcggaa acgcgctgcc gcgtttcgtg
     2281 attgtcacgc tcaagcccgt agtcccgttc gagcgtcgcg cagaggtcag cgagggcgcg
     2341 gtaggcccga tacggctcat ggatggtgtt tcgggtcggg tgaatcttgt tgatggcgat
     2401 atggatgtgc aggttgtcgg tgtcgtgatg cacggcactg acgcgctgat gctcggcgaa
     2461 gccaagccca gcgcagatgc ggtcctcaat cgcgcgcaac gtctccgcgt cgggcttctc
     2521 tcccgcgcgg aagctaacca gcaggtgata ggtcttgtcg gcctcggaac gggtgttgcc
     2581 gtgctgggtc gccatcacct cggccatgac agcgggcagg gtgtttgcct cgcagttcgt
     2641 gacgcgcacg tgacccaggc gctcggtctt gccttgctcg tcggtgatgt acttcaccag
     2701 ctccgcgaag tcgctcttct tgatggagcg catggggacg tgcttggcaa tcacgcgcac
     2761 cccccggccg ttttagcggc taaaaaagtc atggctctgc cctcgggcgg accacgccca
     2821 tcatgacctt gccaagctcg tcctgcttct cttcgatctt cgccagcagg gcgaggatcg
     2881 tggcatcacc gaaccgcgcc gtgcgcgggt cgtcggtgag ccagagtttc agcaggccgc
     2941 ccaggcggcc caggtcgcca ttgatgcggg ccagctcgcg gacgtgctca tagtccacga
     3001 cgcccgtgat tttgtagccc tggccgacgg ccagcaggta ggccgacagg ctcatgccgg
     3061 ccgccgccgc cttttcctca atcgctcttc gttcgtctgg aaggcagtac accttgatag
     3121 gtgggctgcc cttcctggtt ggcttggttt catcagccat ccgcttgccc tcatctgtta
     3181 cgccggcggt agccggccag cctcgcagag caggattccc gttgagcacc gccaggtgcg
     3241 aataagggac agtgaagaag gaacacccgc tcgcgggtgg gcctacttca cctatcctgc
     3301 ccggctgacg ccgttggata caccaaggaa agtctacacg aaccctttgg caaaatcctg
     3361 tatatcgtgc gaaaaaggat ggatataccg aaaaaatcgc tataatgacc ccgaagcagg
     3421 gttatgcagc ggaaaagcgc tgcttccctg ctgttttgtg gaatatctac cgactggaaa
     3481 caggcaaatg caggaaatta ctgaactgag gggacaggcg agagacgatg ccaaagagct
     3541 acaccgacga gctggccgag tgggttgaat cccgcgcggc caagaagcgc cggcgtgatg
     3601 aggctgcggt tgcgttcctg gcggtgaggg cggatgtcga ggcggcgtta gcgtccggct
     3661 atgcgctcgt caccatttgg gagcacatgc gggaaacggg gaaggtcaag ttctcctacg
     3721 agacgttccg ctcgcacgcc aggcggcaca tcaaggccaa gcccgccgat gtgcccgcac
     3781 cgcaggccaa ggctgcggaa cccgcgccgg cacccaagac gccggagcca cggcggccga
     3841 agcagggggg caaggctgaa aagccggccc ccgctgcggc cccgaccggc ttcaccttca
     3901 acccaacacc ggacaaaaag gatccccaat tctcatgttt gacagcttat catcgataag
     3961 ctttaatgcg gtagtttatc acagttaaat tgctaacgca gtcaggcacc gtgtatgaaa
     4021 tctaacaatg cgctcatcgt catcctcggc accgtcaccc tggatgctgt aggcataggc
     4081 ttggttatgc cggtactgcc gggcctcttg cgggatatcg tccattccga cagcatcgcc
     4141 agtcactatg gcgtgctgct agcgctatat gcgttgatgc aatttctatg cgcacccgtt
     4201 ctcggagcac tgtccgaccg ctttggccgc cgcccagtcc tgctcgcttc gctacttgga
     4261 gccactatcg actacgcgat catggcgacc acacccgtcc tgtggatcct ctacgccgga
     4321 cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac
     4381 atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg
     4441 ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt gcatgcacca
     4501 ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt cctaatgcag
     4561 gagtcgcata agggagagcg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
     4621 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
     4681 caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc
     4741 tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct
     4801 caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc
     4861 ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg
     4921 gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc
     4981 atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggatc gctcgcggct
     5041 cttaccagcc taacttcgat cactggaccg ctgatcgtca cggcgattta tgccgcctcg
     5101 gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg ccctatacct tgtctgcctc
     5161 cccgcgttgc gtcgcggtgc atggagccgg gccacctcga cctgaatgga agccggcggc
     5221 acctcgctaa cggattcacc actccaagaa ttggagccaa tcaattcttg cggagaactg
     5281 tgaatgcgca aaccaaccct tggcagaaca tatccatcgc gtccgccatc tccagcagcc
     5341 gcacgcggcg catctcgggc agcgttgggt cctggccggg aattacgccc cgccctgcca
     5401 ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc catcacaaac
     5461 ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt
     5521 gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact
     5581 ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg
     5641 gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg
     5701 ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa
     5761 aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat
     5821 acggaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa
     5881 cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg
     5941 gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg
     6001 ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc
     6061 tgccctatgg gattcacctt tatgttgata agaaataaaa gaaaatgcca ataggatatc
     6121 ggcattttct tttgcgtttt tatttgttaa ctgttaattg tccttgttca aggatgctgt
     6181 ctttgacaac agatgttttc ttgcctttga tgttcagcag gaagcttggc gcaaacgttg
     6241 attgtttgtc tgcgtagaat cctctgtttg tcatatagct tgtaatcacg acattgtttc
     6301 ctttcgcttg aggtacagcg aagtgtgagt aagtaaaggt tacatcgtta ggatcaagat
     6361 ccatttttaa cacaaggcca gttttgttca gcggcttgta tgggccagtt aaagaattag
     6421 aaacataacc aagcatgtaa atatcgttag acgtaatgcc gtcaatcgtc atttttgatc
     6481 cgcgggagtc agtgaacagg taccatttgc cgttcatttt aaagacgttc gcgcgttcaa
     6541 tttcatctgt tactgtgtta gatgcaatca gcggtttcat cacttttttc agtgtgtaat
     6601 catcgtttag ctcaatcata ccgagagcgc cgtttgctaa ctcagccgtg cgttttttat
     6661 cgctttgcag aagtttttga ctttcttgac ggaagaatga tgtgcttttg ccatagtatg
     6721 ctttgttaaa taaagattct tcgccttggt agccatcttc agttccagtg tttgcttcaa
     6781 atactaagta tttgtggcct ttatcttcta cgtagtgagg atctctcagc gtatggttgt
     6841 cgcctgagct gtagttgcct tcatcgatga actgctgtac attttgatac gtttttccgt
     6901 caccgtcaaa gattgattta taatcctcta caccgttgat gttcaaagag ctgtctgatg
     6961 ctgatacgtt aacttgtgca gttgtcagtg tttgtttgcc gtaatgttta ccggagaaat
     7021 cagtgtagaa taaacggatt tttccgtcag atgtaaatgt ggctgaacct gaccattctt
     7081 gtgtttggtc ttttaggata gaatcatttg catcgaattt gtcgctgtct ttaaagacgc
     7141 ggccagcgtt tttccagctg tcaatagaag tttcgccgac tttttgatag aacatgtaaa
     7201 tcgatgtgtc atccgcattt ttaggatctc cggctaatgc aaagacgatg tggtagccgt
     7261 gatagtttgc gacagtgccg tcagcgtttt gtaatggcca gctgtcccaa acgtccaggc
     7321 cttttgcaga agagatattt ttaattgtgg acgaatcgaa ttcaggaact tgatattttt
     7381 catttttttg ctgttcaggg atttgcagca tatcatggcg tgtaatatgg gaaatgccgt
     7441 atgtttcctt atatggcttt tggttcgttt ctttcgcaaa cgcttgagtt gcgcctcctg
     7501 ccagcagtgc ggtagtaaag gttaatactg ttgcttgttt tgcaaacttt ttgatgttca
     7561 tcgttcatgt ctcctttttt atgtactgtg ttagcggtct gcttcttcca gccctcctgt
     7621 ttgaagatgg caagttagtt acgcacaata aaaaaagacc taaaatatgt aaggggtgac
     7681 gccaaagtat acactttgcc ctttacacat tttaggtctt gcctgcttta tcagtaacaa
     7741 acccgcgcga tttacttttc gacctcattc tattagactc tcgtttggat tgcaactggt
     7801 ctattttcct cttttgtttg atagaaaatc ataaaaggat ttgcagacta cgggcctaaa
     7861 gaactaaaaa atctatctgt ttcttttcat tctctgtatt ttttatagtt tctgttgcat
     7921 gggcataaag ttgccttttt aatcacaatt cagaaaatat cataatatct catttcacta
     7981 aataatagtg aacggcaggt atatgtgatg ggttaaaaag gatcgatcct ctagccacgg
     8041 gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact
     8101 ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg
     8161 tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa
     8221 acgcggaagt cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     8281 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     8341 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     8401 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     8461 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     8521 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     8581 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     8641 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     8701 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     8761 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     8821 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     8881 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     8941 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     9001 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     9061 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     9121 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacag
//
