LOCUS       40924_46538        4531 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pwFRT-pheS-gat, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4531)
  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  JOURNAL   Unpublished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 4531)
  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 4531)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4531)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (09-OCT-2009) Molecular Biosciences and Biological Engineering, 
            University of Hawaii at Manoa, 2538 McCarthy Mall, Snyder Hall 207, 
            Honolulu, HI 96822, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..200
                     /label=AmpR promoter
     CDS             201..1058
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1232..1820
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2144..2174
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2289..2356
                     /note="wild type FRT; Flp recognition target"
     protein_bind    complement(2306..2353)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      2393..2434
                     /note="PCS12; rpsL promoter from Burkholderia cenocepacia"
                     /regulatory_class="promoter"
     RBS             2445..2467
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2478..3488
                     /gene="pheS"
                     /label=pheS
                     /note="Phenylalanine--tRNA ligase alpha subunit from 
                     Burkholderia pseudomallei (strain 1710b). Accession#: 
                     Q3JT08"
     CDS             3527..3967
                     /codon_start=1
                     /gene="gat"
                     /product="glyphosate acetyl transferase"
                     /label=gat
                     /note="confers resistance to glyphosate"
                     /protein_id="ADO63837.1"
                     /translation="MIEVKPINAEDTYDLRHRVLRPNQPIEACMFESDLTRSAFHLGGF
                     YGGKLISVASFHQAEHSELQGKKQYQLRGVATLEGYREQKAGSSLVKHAEEILRKRGAD
                     MIWCNARTSASGYYRKLGFSEQGEVFDTPPVGPHILMYKRIT"
     gene            3527..3967
                     /gene="gat"
                     /label=gat
     protein_bind    complement(4023..4070)
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(4137..4153)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt gcatgcctgc aggtcgactc taggatcccc gggtaccgag
     2281 ctcgaattag cttcaaaagc gctctgaagt tcctatactt tctagagaat aggaacttcg
     2341 gaataggaac ttcaagatcc cctgattccc tttgtcaaca gcaatggata tcagccgttg
     2401 acttagttgg tatttccgga atatcatgct gggttccgaa taattttgtt taactttaag
     2461 aaggagatat accatgtatg gacctcgatc agatcgtggc ggatgcccag cagagctttg
     2521 agggcgcggc cgacattacg accttggaaa atgaaaaggc ccgcttcctg ggcaagagcg
     2581 gtgccctcac ggaactcttg aaggggctgg gtaagctgga cccggaaacc cggaagaccg
     2641 agggtgcccg gattaacgtg gccaagcagc aggtggaggc ggccttgaat gcccggcggc
     2701 aggccttggc ggatgccctt ctcaatcagc gcctggcggc tgaagccatt gatgtgaccc
     2761 tccccggtcg cggggcgggc gccggctcgc tgcaccccgt catgcgaacc tgggaacgtg
     2821 tggagcagat tttccgtagc attggcttcg atgtcgcgga tggaccggaa attgagacgg
     2881 attggtataa tttcacggcg ctcaactcgc ccgaaaacca ccccgcacgg tccatgcagg
     2941 acaccttcta tgtggatggt aaggacgccg atgggcggcc cctcctcctc cggacccata
     3001 cctcgcccat gcaggtgcgg tacgcccgca tgaatcgacc cccgattaag gtgattgctc
     3061 ccggtcggac ctatcgggtc gattcggacg ccacccatag ccccatgttt aatcaggtgg
     3121 aggggctctg gattgatgaa aacgtgtcgt ttgccgatct caagggtgcc tacaccgact
     3181 ttctcaagaa atttttcgaa cgagatgaca tcttggtccg tttccgcccg agctacttcc
     3241 ccttcacgga acccagcgcg gaaattgata tgatgtttga acatgggaag aatgctggta
     3301 agtggttgga aatcagcggc agcggtcagg tccaccccac ggtcattcgg aatatgggcc
     3361 tcgacccgga gcgttacatt ggcttcggct tcggttccgg gttggaacgc cttaccatgc
     3421 tccggtatgg tgtccaggac ctccggctct ttttcgaaaa tgacctccgg ttcctccgcc
     3481 agttcgccta aagatccatg ggatcgagct tcgaaaggac aagcatatga tcgaggtcaa
     3541 gcccatcaac gccgaggaca cctacgacct gcggcatcgg gtcctccggc ccaaccagcc
     3601 gatcgaggcg tgcatgttcg aaagcgatct cacgcggagc gccttccacc tgggcgggtt
     3661 ctacgggggc aagctgattt cggtcgcttc gttccaccag gccgagcact cggagcttca
     3721 ggggaagaag cagtaccagc tgcgcggtgt ggccaccctg gagggctacc gggagcagaa
     3781 ggccgggtcg tcgctggtca agcacgccga agagatcctg cggaagcggg gggccgacat
     3841 gatctggtgc aacgcccgga cctcggcctc ggggtactac cggaagctgg ggttcagcga
     3901 gcagggggag gtcttcgaca ccccgcccgt ggggccccac atcctgatgt acaagcggat
     3961 cacctaactc gagctaattc cattgagtaa gtttttaagc acatcagctt caaaagcgct
     4021 ctgaagttcc tatactttct agagaatagg aacttcggaa taggaacttc aagatcccca
     4081 attcgagctc ggtacccggg gatcctagag tcgacctgca ggcatgcaag cttggcactg
     4141 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     4201 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     4261 tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg
     4321 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     4381 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     4441 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     4501 aggttttcac cgtcatcacc gaaacgcgcg a
//
