LOCUS       Exported               10125 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCM410.1145, complete sequence.
ACCESSION   FJ389189
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10125)
  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  JOURNAL   Unpublished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 10125)
  AUTHORS   Marx CJ, 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 10125)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10125
                     /organism="Cloning vector pCM410.1145"
                     /mol_type="other DNA"
                     /db_xref="taxon:646110"
     rep_origin      4..712
                     /label=oriV
                     /note="oriV"
     rep_origin      11..642
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      1105..1693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      1180..1735
                     /label=colE1
                     /note="colE1"
     protein_bind    1981..2002
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     regulatory      2011..2053
                     /regulatory_class="promoter"
                     /note="Plac"
     promoter        2017..2047
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2055..2071
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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"
     regulatory      2206..2306
                     /regulatory_class="promoter"
                     /note="PmxaF"
     primer_bind     2446..2462
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2475..2531
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     CDS             2622..3749
                     /codon_start=1
                     /gene="flhA"
                     /product="glutathione-dependent formaldehyde dehydrogenase"
                     /label=flhA
                     /note="GD-FALDH"
                     /protein_id="ACR45021.1"
                     /translation="MRTRAAVALEAGKPLEVMEVNLEGPKAGEVMVEIKATGICHTDEF
                     TLSGADPEGLFPSILGHEGAGVVVEVGPGVTSVKPGDHVIPLYTPECRQCASCLSGKTN
                     LCTAIRATQGQGLMPDGTSRFSMLDGTPIFHYMGCSTFSNYTVLPEIAVAKVREDAPFD
                     KICYIGCGVTTGIGAVINTAKVEIGAKAVVFGLGGIGLNVLQGLRLAGADMIIGVDLND
                     DKKPMAEHFGMTHFINPKNCENVVQEIVNLTKTPFDQIGGADYSFDCTGNVKVMRDALE
                     CTHRGWGQSIIIGVAPAGAEISTRPFQLVTGRVWKGTAFGGARGRTDVPQIVDWYMDGK
                     IEIDPMITHTLSLDDINKGFDLMHAGESIRSVVLY"
     gene            2622..3749
                     /gene="flhA"
                     /label=flhA
     CDS             3792..4631
                     /codon_start=1
                     /gene="fghA"
                     /product="S-formylglutathione hydrolase"
                     /label=fghA
                     /protein_id="ACR45022.1"
                     /translation="MTLAYETVSENRSFGGIQGVYRHQSQATGTPMTFAIYLPPDARHG
                     KVPVLWYLSGLTCTHENAMTKAGAQEWAAEYGIAVIFPDTSPRGEGVANDETYDLGQGA
                     GFYVDATEAPWAPHFRMWHYVTHELPELVFNNFPLDREAQGITGHSMGGHGALTIAMTF
                     PERYRSVSAFAPIAHPSESDWGRKQFAAYLGDDKAAWKRHDSTILMREKGYPGEVLIDQ
                     GASDQFLDLLKPEALAHAMAERRQPGTFRMQQGYDHSYFFVQSFMADHIRWHAERLG"
     gene            3792..4631
                     /gene="fghA"
                     /label=fghA
     primer_bind     complement(4683..4699)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(5120..5785)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor protein"
                     /label=tetR
                     /note="TetR"
                     /protein_id="ACR45023.1"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGAKKPLLS"
     gene            complement(5120..5785)
                     /gene="tetR"
                     /label=tetR
     CDS             complement(6563..7378)
                     /codon_start=1
                     /gene="aph"
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=aph
                     /note="Aph; kanamycin-resistance protein"
                     /protein_id="ACR45024.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(6563..7378)
                     /gene="aph"
                     /label=aph
     CDS             complement(8050..9198)
                     /codon_start=1
                     /gene="trfA"
                     /product="plasmid replication protein TrfA"
                     /label=trfA
                     /protein_id="ACR45025.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            complement(8050..9198)
                     /gene="trfA"
                     /label=trfA
     oriT            9380..10118
                     /label=origin of transfer
                     /note="origin of transfer"
     CDS             complement(9470..9841)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            9874..9983
                     /note="incP origin of transfer"