LOCUS       40924_40112        4565 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Suicide vector pSHAFT-GFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4565)
  AUTHORS   Shastri S, Spiewak HL, Sofoluwe A, Eidsvaag VA, Asghar AH, Pereira 
            T, Bull EH, Butt AT, Thomas MS.
  TITLE     An efficient system for the generation of marked genetic mutants in 
            members of the genus Burkholderia
  JOURNAL   Plasmid (2016) In press
  PUBMED    27825973
REFERENCE   2  (bases 1 to 4565)
  AUTHORS   Shastri S, Spiewak HL, Pereira T, Sofoluwe AO, Eidsvaag VA, Bull EH,
            Asghar AH, Thomas MS.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 4565)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4565)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid 
            (2016) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-JUN-2016) Department of Infection, Immunity "
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(16..723)
                     /codon_start=1
                     /label=GFP
                     /note="Aequorea victoria green fluorescent protein"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLCYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELY"
     RBS             complement(737..759)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    898..916
                     /label=O end
                     /note="O end"
     protein_bind    complement(928..944)
                     /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)."
     rep_origin      complement(979..1367)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(1988..2356)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(2389..2498)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(3579..4436)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4437..4541)
                     /label=AmpR promoter
ORIGIN
        1 tgcagatcct cagttgtaca gttcatccat gccatgtgta atcccagcag ctgttacaaa
       61 ctcaagaagg accatgtggt ctctcttttc gttgggatct ttcgaaaggg cagattgtgt
      121 ggacaggtaa tggttgtctg gtaaaaggac agggccatcg ccaattggag tattttgttg
      181 ataatggtct gctagttgaa cgcttccatc ttcaatgttg tggcgggtct tgaagttcac
      241 tttgattcca ttcttttgtt tgtctgccat gatgtataca ttgtgtgagt tatagttgta
      301 ttccaatttg tgtcccagaa tgttgccatc ttccttgaag tcaatacctt ttaactcgat
      361 tctattaaca agggtatcac cttcaaactt gacttcagca cgtgtcttgt agttgccgtc
      421 atctttgaag aagatggtcc tttcctgtac ataaccttcg ggcatggcac tcttgaaaaa
      481 gtcatgccgt ttcatatgat ccgggtatct tgaaaagcat tgaacaccat agcacagagt
      541 agtgactagt gttggccatg gaacaggcag tttgccagta gtgcagatga acttcagggt
      601 aagttttccg tatgttgcat caccttcacc ctctccactg acagagaact tgtggccgtt
      661 aacatcacca tctaattcaa caagaattgg gacaactcca gtgaagagtt cttctccttt
      721 gctagccata tgtatatctc cttcttaaag ttaaacaaaa ttatttctac aggggaaggg
      781 cgaattatta tagaaggcga atctgcaaat gtcaaaggcc tttctcgagt tggtaccttt
      841 ctagattaga tctttcccgg gttgtcgaca agcttcggcc gcctaggccg cggccgcact
      901 tgtgtataag agtcagaatt ccacaaattg ttatccgctc acaattccac atgtggaatt
      961 ccacatgtgg aattcccatg tcagccgtta agtgttcctg tgtcactcaa aattgctttg
     1021 agaggctcta agggcttctc agtgcgttac atccctggct tgttgtccac aaccgttaaa
     1081 ccttaaaagc tttaaaagcc ttatatattc ttttttttct tataaaactt aaaaccttag
     1141 aggctattta agttgctgat ttatattaat tttattgttc aaacatgaga gcttagtacg
     1201 tgaaacatga gagcttagta cgttagccat gagagcttag tacgttagcc atgagggttt
     1261 agttcgttaa acatgagagc ttagtacgtt aaacatgaga gcttagtacg tgaaacatga
     1321 gagcttagta cgtactatca acaggttgaa ctgctgatct tcagatcctc tacgccggac
     1381 gcatcgtggc cggatccagc cgaccaggct ttccacgccc gcgtgccgct ccatgtcgtt
     1441 cgcgcggttc tcggaaacgc gctgccgcgt ttcgtgattg tcacgctcaa gcccgtagtc
     1501 ccgttcgagc gtcgcgcaga ggtcagcgag ggcgcggtag gcccgatacg gctcatggat
     1561 ggtgtttcgg gtcgggtgaa tcttgttgat ggcgatatgg atgtgcaggt tgtcggtgtc
     1621 gtgatgcacg gcactgacgc gctgatgctc ggcgaagcca agcccagcgc agatgcggtc
     1681 ctcaatcgcg cgcaacgtct ccgcgtcggg cttctctccc gcgcggaagc taaccagcag
     1741 gtgataggtc ttgtcggcct cggaacgggt gttgccgtgc tgggtcgcca tcacctcggc
     1801 catgacagcg ggcagggtgt ttgcctcgca gttcgtgacg cgcacgtgac ccaggcgctc
     1861 ggtcttgcct tgctcgtcgg tgatgtactt caccagctcc gcgaagtcgc tcttcttgat
     1921 ggagcgcatg gggacgtgct tggcaatcac gcgcaccccc cggccgtttt agcggctaaa
     1981 aaagtcatgg ctctgccctc gggcggacca cgcccatcat gaccttgcca agctcgtcct
     2041 gcttctcttc gatcttcgcc agcagggcga ggatcgtggc atcaccgaac cgcgccgtgc
     2101 gcgggtcgtc ggtgagccag agtttcagca ggccgcccag gcggcccagg tcgccattga
     2161 tgcgggccag ctcgcggacg tgctcatagt ccacgacgcc cgtgattttg tagccctggc
     2221 cgacggccag caggtaggcc gacaggctca tgccggccgc cgccgccttt tcctcaatcg
     2281 ctcttcgttc gtctggaagg cagtacacct tgataggtgg gctgcccttc ctggttggct
     2341 tggtttcatc agccatccgc ttgccctcat ctgttacgcc ggcggtagcc ggccagcctc
     2401 gcagagcagg attcccgttg agcaccgcca ggtgcgaata agggacagtg aagaaggaac
     2461 acccgctcgc gggtgggcct acttcaccta tcctgcccgg ctgacgccgt tggatacacc
     2521 aaggaaagtc tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat
     2581 ataccgaaaa aatcgctata atgaccccga agcagggtta tgcagcggaa aagcgctgct
     2641 tccctgctgt tttgtggaat atctaccgac tggaaacagg caaatgcagg aaattactga
     2701 actgagggga caggcgagag acgatgccaa agagctacac cgacgagctg gccgagtggg
     2761 ttgaatcccg cgcggccaag aagcgccggc gtgatgaggc tgcggttgcg ttcctggcgg
     2821 tgagggcgga tgtcgaggcg gcgttagcgt ccggctatgc gctcgtcacc atttgggagc
     2881 acatgcggga aacggggaag gtcaagttct cctacgagac gttccgctcg cacgccaggc
     2941 ggcacatcaa ggccaagccc gccgatgtgc ccgcaccgca ggccaaggct gcggaacccg
     3001 cgccggcacc caagacgccg gagccacggc ggccgaagca ggggggcaag gctgaaaagc
     3061 cggcccccgc tgcggccccg accggcttca ccttcaaccc aacaccggac aaaaaggatc
     3121 ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc
     3181 tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct
     3241 tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc
     3301 ttgctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     3361 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     3421 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     3481 gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccataaatca
     3541 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3601 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3661 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3721 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3781 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3841 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc
     3901 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3961 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4021 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4081 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4141 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat
     4201 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4261 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4321 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4381 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4441 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4501 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4561 ccacc
//
