LOCUS       40924_24188        8653 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHC36, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8653)
  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  JOURNAL   Unpublished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 8653)
  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 8653)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8653)
  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            "Unpublished"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..8653
                     /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    1054..1659
                     /note="pntAB.1145 allele of Methylobacterium extorquens
                     AM1"
     CDS             complement(2268..2636)
                     /label=traJ
                     /note="oriT-recognizing protein"
     oriT            complement(2669..2778)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     promoter        3415..3443
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             3491..4678
                     /label=TcR
                     /note="tetracycline efflux protein"
     CDS             complement(4862..5518)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             complement(5619..7037)
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     promoter        complement(7038..7483)
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     rep_origin      complement(7895..8483)
                     /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 gagttctgga tcttgcccag ttcctcggcg
     1081 ttgggacggc ggaccttcag gacgaggtcg gcgccggaga gagcctcatc ggcgctcccg
     1141 gcgatcttgg cgccggcggc ttcgtactcg ccatcgggaa cgccggcctt ggcgccggcc
     1201 cccgattgca cgacgacatc ggaccccaga cctatgaact tcttgaccgt ttctgggacg
     1261 gccgcgaccc gtggttctat cgggtccgtc tccgtcaaga cggcaatacg catcagttgc
     1321 tcccgttctt acagccgcct ttcggcgtgc cggtatccga acttgctggg aagttcagat
     1381 ctggaaaggg ccgacgcgag gggtcggccc ggaagtctcg atcctcagct gaggaagaga
     1441 atcgcgagga gtgcggcgaa aacgaaggct tgggccttcc agccgatggc ggggacgaag
     1501 gcaccgacgg ccgcggccac ccatgagacc agcacgccga tgatcgcggt cacccacgct
     1561 tcgtgcacgc cgccgatggc gagagccgtc acccagcaca acaccgtgac ggtaccgatt
     1621 tcgacgaagc ggacgaagcc ccgataggtc tgctcgtggg agctcactag aggatccagc
     1681 cgaccaggct ttccacgccc gcgtgccgct ccatgtcgtt cgcgcggttc tcggaaacgc
     1741 gctgccgcgt ttcgtgattg tcacgctcaa gcccgtagtc ccgttcgagc gtcgcgcaga
     1801 ggtcagcgag ggcgcggtag gcccgatacg gctcatggat ggtgtttcgg gtcgggtgaa
     1861 tcttgttgat ggcgatatgg atgtgcaggt tgtcggtgtc gtgatgcacg gcactgacgc
     1921 gctgatgctc ggcgaagcca agcccagcgc agatgcggtc ctcaatcgcg cgcaacgtct
     1981 ccgcgtcggg cttctctccc gcgcggaagc taaccagcag gtgataggtc ttgtcggcct
     2041 cggaacgggt gttgccgtgc tgggtcgcca tcacctcggc catgacagcg ggcagggtgt
     2101 ttgcctcgca gttcgtgacg cgcacgtgac ccaggcgctc ggtcttgcct tgctcgtcgg
     2161 tgatgtactt caccagctcc gcgaagtcgc tcttcttgat ggagcgcatg gggacgtgct
     2221 tggcaatcac gcgcaccccc cggccgtttt agcggctaaa aaagtcatgg ctctgccctc
     2281 gggcggacca cgcccatcat gaccttgcca agctcgtcct gcttctcttc gatcttcgcc
     2341 agcagggcga ggatcgtggc atcaccgaac cgcgccgtgc gcgggtcgtc ggtgagccag
     2401 agtttcagca ggccgcccag gcggcccagg tcgccattga tgcgggccag ctcgcggacg
     2461 tgctcatagt ccacgacgcc cgtgattttg tagccctggc cgacggccag caggtaggcc
     2521 gacaggctca tgccggccgc cgccgccttt tcctcaatcg ctcttcgttc gtctggaagg
     2581 cagtacacct tgataggtgg gctgcccttc ctggttggct tggtttcatc agccatccgc
     2641 ttgccctcat ctgttacgcc ggcggtagcc ggccagcctc gcagagcagg attcccgttg
     2701 agcaccgcca ggtgcgaata agggacagtg aagaaggaac acccgctcgc gggtgggcct
     2761 acttcaccta tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc tacacgaacc
     2821 ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata
     2881 atgaccccga agcagggtta tgcagcggaa aagcgctgct tccctgctgt tttgtggaat
     2941 atctaccgac tggaaacagg caaatgcagg aaattactga actgagggga caggcgagag
     3001 acgatgccaa agagctacac cgacgagctg gccgagtggg ttgaatcccg cgcggccaag
     3061 aagcgccggc gtgatgaggc tgcggttgcg ttcctggcgg tgagggcgga tgtcgaggcg
     3121 gcgttagcgt ccggctatgc gctcgtcacc atttgggagc acatgcggga aacggggaag
     3181 gtcaagttct cctacgagac gttccgctcg cacgccaggc ggcacatcaa ggccaagccc
     3241 gccgatgtgc ccgcaccgca ggccaaggct gcggaacccg cgccggcacc caagacgccg
     3301 gagccacggc ggccgaagca ggggggcaag gctgaaaagc cggcccccgc tgcggccccg
     3361 accggcttca ccttcaaccc aacaccggac aaaaaggatc cccaattctc atgtttgaca
     3421 gcttatcatc gataagcttt aatgcggtag tttatcacag ttaaattgct aacgcagtca
     3481 ggcaccgtgt atgaaatcta acaatgcgct catcgtcatc ctcggcaccg tcaccctgga
     3541 tgctgtaggc ataggcttgg ttatgccggt actgccgggc ctcttgcggg atatcgtcca
     3601 ttccgacagc atcgccagtc actatggcgt gctgctagcg ctatatgcgt tgatgcaatt
     3661 tctatgcgca cccgttctcg gagcactgtc cgaccgcttt ggccgccgcc cagtcctgct
     3721 cgcttcgcta cttggagcca ctatcgacta cgcgatcatg gcgaccacac ccgtcctgtg
     3781 gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg
     3841 cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag
     3901 cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat
     3961 ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg
     4021 ctgcttccta atgcaggagt cgcataaggg agagcgtcga ccgatgccct tgagagcctt
     4081 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     4141 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg tcattttcgg
     4201 cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat
     4261 cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa
     4321 gcaggccatt atcgccggca tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc
     4381 gacgcgaggc tggatggcct tccccattat gattcttctc gcttccggcg gcatcgggat
     4441 gcccgcgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg gacagcttca
     4501 aggatcgctc gcggctctta ccagcctaac ttcgatcact ggaccgctga tcgtcacggc
     4561 gatttatgcc gcctcggcga gcacatggaa cgggttggca tggattgtag gcgccgccct
     4621 ataccttgtc tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca cctcgacctg
     4681 aatggaagcc ggcggcacct cgctaacgga ttcaccactc caagaattgg agccaatcaa
     4741 ttcttgcgga gaactgtgaa tgcgcaaacc aacccttggc agaacatatc catcgcgtcc
     4801 gccatctcca gcagccgcac gcggcgcatc tcgggcagcg ttgggtcctg gccgggaatt
     4861 acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat
     4921 ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc
     4981 cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca
     5041 cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct
     5101 caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat
     5161 atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt
     5221 cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac
     5281 cgtctttcat tgccatacgg aattccggat gagcattcat caggcgggca agaatgtgaa
     5341 taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat
     5401 ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt
     5461 ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt
     5521 tagcttcctt agctcctgcc ctatgggatt cacctttatg ttgataagaa ataaaagaaa
     5581 atgccaatag gatatcggca ttttcttttg cgtttttatt tgttaactgt taattgtcct
     5641 tgttcaagga tgctgtcttt gacaacagat gttttcttgc ctttgatgtt cagcaggaag
     5701 cttggcgcaa acgttgattg tttgtctgcg tagaatcctc tgtttgtcat atagcttgta
     5761 atcacgacat tgtttccttt cgcttgaggt acagcgaagt gtgagtaagt aaaggttaca
     5821 tcgttaggat caagatccat ttttaacaca aggccagttt tgttcagcgg cttgtatggg
     5881 ccagttaaag aattagaaac ataaccaagc atgtaaatat cgttagacgt aatgccgtca
     5941 atcgtcattt ttgatccgcg ggagtcagtg aacaggtacc atttgccgtt cattttaaag
     6001 acgttcgcgc gttcaatttc atctgttact gtgttagatg caatcagcgg tttcatcact
     6061 tttttcagtg tgtaatcatc gtttagctca atcataccga gagcgccgtt tgctaactca
     6121 gccgtgcgtt ttttatcgct ttgcagaagt ttttgacttt cttgacggaa gaatgatgtg
     6181 cttttgccat agtatgcttt gttaaataaa gattcttcgc cttggtagcc atcttcagtt
     6241 ccagtgtttg cttcaaatac taagtatttg tggcctttat cttctacgta gtgaggatct
     6301 ctcagcgtat ggttgtcgcc tgagctgtag ttgccttcat cgatgaactg ctgtacattt
     6361 tgatacgttt ttccgtcacc gtcaaagatt gatttataat cctctacacc gttgatgttc
     6421 aaagagctgt ctgatgctga tacgttaact tgtgcagttg tcagtgtttg tttgccgtaa
     6481 tgtttaccgg agaaatcagt gtagaataaa cggatttttc cgtcagatgt aaatgtggct
     6541 gaacctgacc attcttgtgt ttggtctttt aggatagaat catttgcatc gaatttgtcg
     6601 ctgtctttaa agacgcggcc agcgtttttc cagctgtcaa tagaagtttc gccgactttt
     6661 tgatagaaca tgtaaatcga tgtgtcatcc gcatttttag gatctccggc taatgcaaag
     6721 acgatgtggt agccgtgata gtttgcgaca gtgccgtcag cgttttgtaa tggccagctg
     6781 tcccaaacgt ccaggccttt tgcagaagag atatttttaa ttgtggacga atcgaattca
     6841 ggaacttgat atttttcatt tttttgctgt tcagggattt gcagcatatc atggcgtgta
     6901 atatgggaaa tgccgtatgt ttccttatat ggcttttggt tcgtttcttt cgcaaacgct
     6961 tgagttgcgc ctcctgccag cagtgcggta gtaaaggtta atactgttgc ttgttttgca
     7021 aactttttga tgttcatcgt tcatgtctcc ttttttatgt actgtgttag cggtctgctt
     7081 cttccagccc tcctgtttga agatggcaag ttagttacgc acaataaaaa aagacctaaa
     7141 atatgtaagg ggtgacgcca aagtatacac tttgcccttt acacatttta ggtcttgcct
     7201 gctttatcag taacaaaccc gcgcgattta cttttcgacc tcattctatt agactctcgt
     7261 ttggattgca actggtctat tttcctcttt tgtttgatag aaaatcataa aaggatttgc
     7321 agactacggg cctaaagaac taaaaaatct atctgtttct tttcattctc tgtatttttt
     7381 atagtttctg ttgcatgggc ataaagttgc ctttttaatc acaattcaga aaatatcata
     7441 atatctcatt tcactaaata atagtgaacg gcaggtatat gtgatgggtt aaaaaggatc
     7501 gatcctctag ccacgggtgc gcatgatcgt gctcctgtcg ttgaggaccc ggctaggctg
     7561 gcggggttgc cttactggtt agcagaatga atcaccgata cgcgagcgaa cgtgaagcga
     7621 ctgctgctgc aaaacgtctg cgacctgagc aacaacatga atggtcttcg gtttccgtgt
     7681 ttcgtaaagt ctggaaacgc ggaagtcagc gctcttccgc ttcctcgctc actgactcgc
     7741 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     7801 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     7861 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7921 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     7981 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     8041 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     8101 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     8161 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     8221 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     8281 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     8341 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     8401 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     8461 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     8521 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     8581 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     8641 cttggtctga cag
//
