LOCUS       40924_24208        8855 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Promoter-probe vector pHC42, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8855)
  AUTHORS   Chou H-H., Berthet J, Marx CJ.
  TITLE     Parallel transpositions of an insertion sequence drove adaptation of
            Methylobacterium in a metal-limited environment
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8855)
  AUTHORS   Chou H-H.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-APR-2008) Organismic and Evolutionary Biology, Harvard
            University, 16 Divinity Avenue, Biolab 3079, Cambridge, MA 02138, 
            USA
REFERENCE   3  (bases 1 to 8855)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8855)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-APR-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..8855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      3..712
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      1105..1693
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    1981..2002
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2017..2047
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2055..2071
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2079..2095
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2123..2836)
                     /label=GFPuv
                     /note="GFP variant optimized for excitation by UV light"
     regulatory      complement(2837..2851)
                     /note="derived from Methylobacterium extorquens AM1 fae
                     gene"
                     /regulatory_class="ribosome_binding_site"
     terminator      complement(3026..3053)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(3145..3231)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     complement(3413..3429)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3874..4515)
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
     CDS             complement(5296..6108)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             complement(6783..7928)
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
     CDS             complement(8200..8571)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(8604..8713)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
ORIGIN
        1 gaccctttcc gacgctcacc gggctggttg ccctcgccgc tgggctggcg gccgtctatg
       61 gccctgcaaa cgcgccagaa acgccgtcga agccgtgtgc gagacaccgc ggccgccggc
      121 gttgtggata cctcgcggaa aacttggccc tcactgacag atgaggggcg gacgttgaca
      181 cttgaggggc cgactcaccc ggcgcggcgt tgacagatga ggggcaggct cgatttcggc
      241 cggcgacgtg gagctggcca gcctcgcaaa tcggcgaaaa cgcctgattt tacgcgagtt
      301 tcccacagat gatgtggaca agcctgggga taagtgccct gcggtattga cacttgaggg
      361 gcgcgactac tgacagatga ggggcgcgat ccttgacact tgaggggcag agtgctgaca
      421 gatgaggggc gcacctattg acatttgagg ggctgtccac aggcagaaaa tccagcattt
      481 gcaagggttt ccgcccgttt ttcggccacc gctaacctgt cttttaacct gcttttaaac
      541 caatatttat aaaccttgtt tttaaccagg gctgcgccct gtgcgcgtga ccgcgcacgc
      601 cgaagggggg tgccccccct tctcgaaccc tcccggcccg ctaacgcggg cctcccatcc
      661 ccccaggggc tgcgcccctc ggccgcgaac ggcctcaccc caaaaatggc agccaagctg
      721 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
      781 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
      841 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
      901 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
      961 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     1021 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1081 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1141 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1201 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     1261 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1321 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1381 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1441 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1501 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1561 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1621 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1681 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1741 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     1801 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     1861 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     1921 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     1981 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     2041 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     2101 ttacgccaag cttgcatgct tatttgtaga gctcatccat gccatgtgta atcccagcag
     2161 cagttacaaa ctcaagaagg accatgtggt cacgcttttc gttgggatct ttcgaaaggg
     2221 cagattgtgt cgacaggtaa tggttgtctg gtaaaaggac agggccatcg ccaattggag
     2281 tattttgttg ataatggtct gctagttgaa cggatccatc ttcaatgttg tggcgaattt
     2341 tgaagttagc tttgattcca ttcttttgtt tgtctgccgt gatgtataca ttgtgtgagt
     2401 tatagttgta ctcgagtttg tgtccgagaa tgtttccatc ttctttaaaa tcaatacctt
     2461 ttaactcgat acgattaaca agggtatcac cttcaaactt gacttcagca cgcgtcttgt
     2521 agttcccgtc atctttgaaa gatatagtgc gttcctgtac ataaccttcg ggcatggcac
     2581 tcttgaaaaa gtcatgccgt ttcatatgat ccggataacg ggaaaagcat tgaacaccat
     2641 aagagaaagt agtgacaagt gttggccatg gaacaggtag ttttccagta gtgcaaataa
     2701 atttaagggt aagttttccg tatgttgcat caccttcacc ctctccactg acagaaaatt
     2761 tgtgcccatt aacatcacca tctaattcaa caagaattgg gacaactcca gtgaaaagtt
     2821 cttctccttt actcattcgg ggtctctccc tgctagcggg cccggatccg atatcttaag
     2881 atcttccgtg cagtttaagc cgaattgatc cccgggtacc gagctcgaat tccctgcttt
     2941 cctgatgcaa aaacgaggct agtttaccgt atctgtgggg ggatggcttg tagatatgac
     3001 gacaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttaat
     3061 ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcgc
     3121 aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca
     3181 acagataaaa cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgcctggc
     3241 agttccctac tctcgcatgg ggagacccca cactaccatc ggcgctacgg cgtttcactt
     3301 ctgagttcgg catggggtca ggtgggacca ccgcgctact gccgccaggc aaattctgtt
     3361 ttatcagacc gcttctgcgt tctgatttaa tctgtatcag gctgaaaaat tcactggccg
     3421 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     3481 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     3541 aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc
     3601 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     3661 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3721 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3781 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     3841 aggttaatgt catgataata atggtttctt agcacccttt ctcggtcctt caacgttcct
     3901 gacaacgagc ctccttttcg ccaatccatc gacaatcacc gcgagtccct gctcgaacgc
     3961 tgcgtccgga ccggcttcgt cgaaggcgtc tatcgcggcc cgcaacagcg gcgagagcgg
     4021 agcctgttca acggtgccgc cgcgctcgcc ggcatcgctg tcgccggcct gctcctcaag
     4081 cacggcccca acagtgaagt agctgattgt catcagcgca ttgacggcgt ccccggccga
     4141 aaaacccgcc tcgcagagga agcgaagctg cgcgtcggcc gtttccatct gcggtgcgcc
     4201 cggtcgcgtg ccggcatgga tgcgcgcgcc atcgcggtag gcgagcagcg cctgcctgaa
     4261 gctgcgggca ttcccgatca gaaatgagcg ccagtcgtcg tcggctctcg gcaccgaatg
     4321 cgtatgattc tccgccagca tggcttcggc cagtgcgtcg agcagcgccc gcttgttcct
     4381 gaagtgccag taaagcgccg gctgctgaac ccccaaccgt tccgccagtt tgcgtgtcgt
     4441 cagaccgtct acgccgacct cgttcaacag gtccagggcg gcacggatca ctgtattcgg
     4501 ctgcaacttt gtcatgattg acactttatc actgataaac ataatatgtc caccaactta
     4561 tcagtgataa agaatccgcg cgttcaatcg gaccagcgga ggctggtccg gaggccagac
     4621 gtgaaaccca acatacccct gatcgtaatt ctgagcactg tcgcgctcga cgctgtcggc
     4681 atcggcctga ttatgccggt gctgccgggc ctcctgcgcg atctggttca ctcgaacgac
     4741 gtcaccgccc actatggcat tctgctggcg ctgtatgcgt tggtgcaatt tgcctgcgca
     4801 cctgtgctgg gcgcgctgtc ggatcgtttc gggcggcggc caatcttgct cgtctcgctg
     4861 gccggcgcca ctgtcgacta cgccatcatg gcgacagcgc ctttcctttg ggttctctat
     4921 atcgggcgga tcgtggccgg catcaccggg gcgactgggg cggtagccgg cgcttatatt
     4981 gccgatgacc tgcagggggg ggggggcgct gaggtctgcc tcgtgaagaa ggtgttgctg
     5041 actcatacca ggcctgaatc gccccatcat ccagccagaa agtgagggag ccacggttga
     5101 tgagagcttt gttgtaggtg gaccagttgg tgattttgaa cttttgcttt gccacggaac
     5161 ggtctgcgtt gtcgggaaga tgcgtgatct gatccttcaa ctcagcaaaa gttcgattta
     5221 ttcaacaaag ccgccgtccc gtcaagtcag cgtaatgctc tgccagtgtt acaaccaatt
     5281 aaccaattct gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc
     5341 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     5401 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     5461 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     5521 atgagtgacg actgaatccg gtgagaatgg caaaagctta tgcatttctt tccagacttg
     5581 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt
     5641 cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca
     5701 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc
     5761 tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag
     5821 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     5881 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     5941 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc
     6001 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     6061 atttaatcgc ggcctcgagc aagacgtttc ccgttgaata tggctcataa caccccttgt
     6121 attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc
     6181 aatgtaacat cagagatttt gagacacaac gtggctttcc cccccccccc tgcaggtccg
     6241 acacggggat ggatggcgtt cccgatcatg gtcctgcttg cttcgggtgg catcggaatg
     6301 ccggcgctgc aagcaatgtt gtccaggcag gtggatgagg aacgtcaggg gcagctgcaa
     6361 ggctcactgg cggcgctcac cagcctgacc tcgatcgtcg gacccctcct cttcacggcg
     6421 atctatgcgg cttctataac aacgtggaac gggtgggcat ggattgcagg cgctgccctc
     6481 tacttgctct gcctgccggc gctgcgtcgc gggctttgga gcggcgcagg gcaacgagcc
     6541 gatcgctgat cgtggaaacg ataggcctat gccatgcggg tcaaggcgac ttccggcaag
     6601 ctatacgcgc cctagaattg tcaattttaa tcctctgttt atcggcagtt cgtagagcgc
     6661 gccgtgcgtc ccgagcgata ctgagcgaag caagtgcgtc gagcagtgcc cgcttgttcc
     6721 tgaaatgcca gtaaagcgct ggctgctgaa cccccagccg gaactgaccc cacaaggccc
     6781 tagcgtttgc aatgcaccag gtcatcattg acccaggcgt gttccaccag gccgctgcct
     6841 cgcaactctt cgcaggcttc gccgacctgc tcgcgccact tcttcacgcg ggtggaatcc
     6901 gatccgcaca tgaggcggaa ggtttccagc ttgagcgggt acggctcccg gtgcgagctg
     6961 aaatagtcga acatccgtcg ggccgtcggc gacagcttgc ggtacttctc ccatatgaat
     7021 ttcgtgtagt ggtcgccagc aaacagcacg acgatttcct cgtcgatcag gacctggcaa
     7081 cgggacgttt tcttgccacg gtccaggacg cggaagcggt gcagcagcga caccgattcc
     7141 aggtgcccaa cgcggtcgga cgtgaagccc atcgccgtcg cctgtaggcg cgacaggcat
     7201 tcctcggcct tcgtgtaata ccggccattg atcgaccagc ccaggtcctg gcaaagctcg
     7261 tagaacgtga aggtgatcgg ctcgccgata ggggtgcgct tcgcgtactc caacacctgc
     7321 tgccacacca gttcgtcatc gtcggcccgc agctcgacgc cggtgtaggt gatcttcacg
     7381 tccttgttga cgtggaaaat gaccttgttt tgcagcgcct cgcgcgggat tttcttgttg
     7441 cgcgtggtga acagggcaga gcgggccgtg tcgtttggca tcgctcgcat cgtgtccggc
     7501 cacggcgcaa tatcgaacaa ggaaagctgc atttccttga tctgctgctt cgtgtgtttc
     7561 agcaacgcgg cctgcttggc ctcgctgacc tgttttgcca ggtcctcgcc ggcggttttt
     7621 cgcttcttgg tcgtcatagt tcctcgcgtg tcgatggtca tcgacttcgc caaacctgcc
     7681 gcctcctgtt cgagacgacg cgaacgctcc acggcggccg atggcgcggg cagggcaggg
     7741 ggagccagtt gcacgctgtc gcgctcgatc ttggccgtag cttgctggac catcgagccg
     7801 acggactgga aggtttcgcg gggcgcacgc atgacggtgc ggcttgcgat ggtttcggca
     7861 tcctcggcgg aaaaccccgc gtcgatcagt tcttgcctgt atgccttccg gtcaaacgtc
     7921 cgattcattc accctccttg cgggattgcc ccgactcacg ccggggcaat gtgcccttat
     7981 tcctgatttg acccgcctgg tgccttggtg tccagataat ccaccttatc ggcaatgaag
     8041 tcggtcccgt agaccgtctg gccgtccttc tcgtacttgg tattccgaat cttgccctgc
     8101 acgaatacca gctccgcgaa gtcgctcttc ttgatggagc gcatggggac gtgcttggca
     8161 atcacgcgca ccccccggcc gttttagcgg ctaaaaaagt catggctctg ccctcgggcg
     8221 gaccacgccc atcatgacct tgccaagctc gtcctgcttc tcttcgatct tcgccagcag
     8281 ggcgaggatc gtggcatcac cgaaccgcgc cgtgcgcggg tcgtcggtga gccagagttt
     8341 cagcaggccg cccaggcggc ccaggtcgcc attgatgcgg gccagctcgc ggacgtgctc
     8401 atagtccacg acgcccgtga ttttgtagcc ctggccgacg gccagcaggt aggcctacag
     8461 gctcatgccg gccgccgccg ccttttcctc aatcgctctt cgttcgtctg gaaggcagta
     8521 caccttgata ggtgggctgc ccttcctggt tggcttggtt tcatcagcca tccgcttgcc
     8581 ctcatctgtt acgccggcgg tagccggcca gcctcgcaga gcaggattcc cgttgagcac
     8641 cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg ggcctacttc
     8701 acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac gaaccctttg
     8761 gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg ctataatgac
     8821 cccgaagcag ggttatgcag cggaaaagat ccgtc
//
