LOCUS       Exported                5321 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector Mini-Tn4001PSpuro, complete sequence.
ACCESSION   FJ872396
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5321)
  AUTHORS   Algire MA, Lartigue C, Thomas DW, Assad-Garcia N, Glass JI, Merryman
            C.
  TITLE     New selectable marker for manipulating the simple genomes of 
            Mycoplasma species
  JOURNAL   Antimicrob. Agents Chemother. 53 (10), 4429-4432 (2009)
  PUBMED    19687239
REFERENCE   2  (bases 1 to 5321)
  AUTHORS   Merryman C.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-MAR-2009) Synthetic Biology, J. Craig Venter 
            Institute, 9704 Medical Center Dr, Rockville, MD 20850, USA
REFERENCE   3  (bases 1 to 5321)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5321
                     /organism="Cloning vector Mini-Tn4001PSpuro"
                     /mol_type="other DNA"
                     /db_xref="taxon:640477"
     CDS             complement(1..600)
                     /codon_start=1
                     /gene="pac"
                     /product="puromycin N-acetyltransferase"
                     /label=pac
                     /note="puromycin resistance; codon optimized for Mycoplasma
                     genitalium"
                     /protein_id="ACR82772.1"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     gene            complement(1..600)
                     /gene="pac"
                     /label=pac
     CDS             1105..2277
                     /codon_start=1
                     /gene="tnp"
                     /product="transposase"
                     /label=tnp
                     /protein_id="ACR82773.1"
                     /translation="MTQVHFTLKSEEIQSIIEYSVKDDVSKNILTTVFNQLMENQRTEY
                     IQAKEYERTENRQSQRNGYYERSFTTRVGTLELKVPRTRDGHFSPTVFERYQRNEKALM
                     ASMLEMYVSGVSTRKVSKIVEELCGKSVSKSFVSSLTEQLEPMVNEWQNRLLSEKNYPY
                     LMTDVLYIKVREENRVLSKSCHIAIGITKDGDREIIGFMIQSGESEETWTTFFEYLKER
                     GLQGTELVISDAHKGLVSAIRKSFTNVSWQRCQVHFLRNIFTTIPKKNSKSFREAVKGI
                     FKFTDINLAREAKNRLIHDYIDQPKYSKACASLDDGFEDAFQYTVQGNSHNRLKSTNLI
                     ERLNQEVRRREKIIRIFPNQTSANRLIGAVLMDLHDEWIYSSRKYINFDK"
     gene            1105..2277
                     /gene="tnp"
                     /label=tnp
     promoter        complement(2298..2316)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2323..2339)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2481..2936
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2962..3066
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3067..3927
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance"
                     /protein_id="ACR82774.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            3067..3927
                     /gene="bla"
                     /label=bla
     rep_origin      4098..4686
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4974..4995
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5010..5040
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5048..5064
                     /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     5072..5088
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5109..5127
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"