LOCUS       Exported                3323 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAM3G, complete sequence.
ACCESSION   GU074522
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3323)
  AUTHORS   Kang Y, Norris MH, Hoang TT.
  TITLE     Knock-out and pull-out recombineering protocols for natural 
            transformable Burkholderia thailandensis and Burkholderia 
            pseudomallei
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3323)
  AUTHORS   Kang Y, Norris MH, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-OCT-2009) Molecular Biosciences and Biological 
            Engineering, University of Hawaii at Manoa, 2538 McCarthy Mall, 
            Snyder Hall 207, Honolulu, HI 96822, USA
REFERENCE   3  (bases 1 to 3323)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3323
                     /organism="Cloning vector pAM3G"
                     /mol_type="other DNA"
                     /label=vector based on glyphosate resistance
                     /note="vector based on glyphosate resistance"
                     /db_xref="taxon:908463"
     rep_origin      12..363
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from 
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     rep_origin      15..370
                     /note="ori1600; broad host range origin of replication"
     CDS             377..1210
                     /codon_start=1
                     /gene="rep"
                     /product="replication protein"
                     /label=rep
                     /note="recognizes ori1600 for broad-host-range replication"
                     /protein_id="ADO63832.1"
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     gene            377..1210
                     /gene="rep"
                     /label=rep
     CDS             complement(1207..1461)
                     /codon_start=1
                     /gene="lacZa"
                     /product="LacZa"
                     /label=lacZa
                     /note="alpha subunit for blue-white selection"
                     /protein_id="ADO63833.1"
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLKGRPGRGGHGL"
     gene            complement(1207..1461)
                     /gene="lacZa"
                     /label=lacZa
     primer_bind     1371..1387
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1388..1444
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1457..1473)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(1474..1659)
                     /regulatory_class="promoter"
                     /note="Plac; lac-promoter"
     protein_bind    1481..1497
                     /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)."
     promoter        complement(1505..1535)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1550..1571
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     oriT            1803..2070
     oriT            1955..2063
                     /note="incP origin of transfer"
     rep_origin      complement(2131..2719)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      2143..2668
                     /label=ColE1 origin of replication
                     /note="ColE1 origin of replication"
     CDS             complement(2803..3243)
                     /codon_start=1
                     /gene="gat"
                     /product="glyphosate acetyl transferase"
                     /label=gat
                     /note="confers resistance to glyphosate"
                     /protein_id="ADO63834.1"
                     /translation="MIEVKPINAEDTYDLRHRVLRPNQPIEACMFESDLTRSAFHLGGF
                     YGGKLISVASFHQAEHSELQGKKQYQLRGVATLEGYREQKAGSSLVKHAEEILRKRGAD
                     MIWCNARTSASGYYRKLGFSEQGEVFDTPPVGPHILMYKRIT"
     gene            complement(2803..3243)
                     /gene="gat"
                     /label=gat
     regulatory      3282..3323
                     /regulatory_class="promoter"
                     /note="PCS12; rpsL promoter from Burkholderia cenocepacia"