LOCUS       40924_21593        6157 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGFPbglS, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6157)
  AUTHORS   Bisicchia P, Botella E, Devine KM.
  TITLE     Suite of novel vectors for ectopic insertion of GFP, CFP and IYFP 
            transcriptional fusions in single copy at the amyE and bglS loci in 
            Bacillus subtilis
  JOURNAL   Plasmid 64 (3), 143-149 (2010)
  PUBMED    20600285
REFERENCE   2  (bases 1 to 6157)
  AUTHORS   Bisicchia P.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-MAY-2010) Trinity College Dublin, 13 Meades Terrace, 
            Dublin 2, Ireland
REFERENCE   3  (bases 1 to 6157)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6157)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid"; 
            date: "2010"; volume: "64"; issue: "3"; pages: "143-149"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-MAY-2010) Trinity College Dublin, 13 Meades Terrace, Dublin 2, 
            Ireland"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(6..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     677..693
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_recomb     695..1121
                     /label=bglS front
                     /note="bglS front"
     misc_feature    1134..1169
                     /label=LIC site
                     /note="LIC site"
     CDS             1170..1883
                     /codon_start=1
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1906..1999
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2571..3362
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     primer_bind     complement(3522..3538)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_recomb     3541..3938
                     /label=bglS back
                     /note="bglS back"
     promoter        complement(3972..3990)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4011..4027)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4035..4051)
                     /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)."
     promoter        complement(4059..4089)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4104..4125)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4413..5001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5175..6032)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6033..6137)
                     /label=AmpR promoter
ORIGIN
        1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
       61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
      181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
      241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggtggc ggccgctcta gaactagtgg atccgcttca ctattatcgg ttcgtcaccc
      721 acttaaagtt tttcgcgcag cgtctattta acggcactca catggaaagc caggacgatt
      781 ttttgctgga tacagtgaaa gaaaagtatc atcgcgcgta tgaatgcacg aagaaaatcc
      841 aaacctacat tgagcgggag tatgagcaca agctcacaag tgacgagctg ctgtatttaa
      901 ccattcacat agaaagggta gttaaacaag cataatgaga gcgctgacat ttgtgtttcc
      961 ttgtgttcac tttttcttac attcacatat gaaaatggta ggattgttac tgataaagca
     1021 ggcaaaacct aaattgcaat gagtgcggat catctctgtc tgtgctgatg gtaatttagg
     1081 tttttatttt tttcagaggg aagatgatga tagttacagg actgcaggaa ttcttttacc
     1141 gcgggctttc ccgggaagga ggaactacta tgcgtaaagg agaagaactt ttcactggag
     1201 ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg
     1261 gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg
     1321 gaaaactacc tgttccatgg ccaacacttg tcactacttt cggttatggt gttcaatgct
     1381 ttgcgagata cccagatcat atgaaacggc atgacttttt caagagtgcc atgcccgaag
     1441 gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag acacgtgctg
     1501 aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt attgatttta
     1561 aagaagatgg aaacattctt ggacacaaat tggaatacaa ctataactca cacaatgtat
     1621 acatcatggc agacaaacaa aagaatggaa tcaaagttaa cttcaaaatt agacacaaca
     1681 ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg
     1741 gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc
     1801 ccaacgaaaa gagagaccac atggtccttc ttgagtttgt aacagctgct gggattacac
     1861 atggcatgga tgaactatac aaataagctt aattagctga gcttggactc ctgttgatag
     1921 atccagtaat gacctcagaa ctccatctgg atttgttcag aacgctcggt tgccgccggg
     1981 cgttttttat tggtgagaag tcgactacag aaccgaattc gatatcaaag cttatcgata
     2041 aacccagcga accatttgag gtgataggta agattatacc gaggtatgaa aacgagaatt
     2101 ggacctttac agaattactc tatgaagcgc catatttaaa aagctaccaa gacgaagagg
     2161 atgaagagga tgaggaggca gattgccttg aatatattga caatactgat aagataatat
     2221 atcttttata tagaagatat cgccgtatgt aaggatttca gggggcaagg cataggcagc
     2281 gcgcttatca atatatctat agaatgggca aagcataaaa acttgcatgg actaatgctt
     2341 gaaacccagg acaataacct tatagcttgt aaattctatc ataattgtgg tttcaaaatc
     2401 ggctccgtcg atactatgtt atacgccaac tttcaaaaca actttgaaaa agctgttttc
     2461 tggtatttaa ggttttagaa tgcaaggaac agtgaattgg agttcgtctt gttataatta
     2521 gcttcttggg gtatctttaa atactgtaga aaagaggaag gaaataataa atggctaaaa
     2581 tgagaatatc accggaattg aaaaaactga tcgaaaaata ccgctgcgta aaagatacgg
     2641 aaggaatgtc tcctgctaag gtatataagc tggtgggaga aaatgaaaac ctatatttaa
     2701 aaatgacgga cagccggtat aaagggacca cctatgatgt ggaacgggaa aaggacatga
     2761 tgctatggct ggaaggaaag ctgcctgttc caaaggtcct gcactttgaa cggcatgatg
     2821 gctggagcaa tctgctcatg agtgaggccg atggcgtcct ttgctcggaa gagtatgaag
     2881 atgaacaaag ccctgaaaag attatcgagc tgtatgcgga gtgcatcagg ctctttcact
     2941 ccatcgacat atcggattgt ccctatacga atagcttaga cagccgctta gccgaattgg
     3001 attacttact gaataacgat ctggccgatg tggattgcga aaactgggaa gaagacactc
     3061 catttaaaga tccgcgcgag ctgtatgatt ttttaaagac ggaaaagccc gaagaggaac
     3121 ttgtcttttc ccacggcgac ctgggagaca gcaacatctt tgtgaaagat ggcaaagtaa
     3181 gtggctttat tgatcttggg agaagcggca gggcggacaa gtggtatgac attgccttct
     3241 gcgtccggtc gatcagggag gatatcgggg aagaacagta tgtcgagcta ttttttgact
     3301 tactggggat caagcctgat tgggagaaaa taaaatatta tattttactg gatgaattgt
     3361 tttagtacct agatttagat gtctaaaaag ctttaactac aagcttttta gacatctaat
     3421 cttttctgaa gtacatccgc aactgtccat actctgatgt tttatatctt ttctaaaagt
     3481 tcgctagata ggggtcccga gcgcctacga ggaatttgta tcgataccgt cgacctcgag
     3541 cttcacttac acaggtccaa cagatggaac tccttgggat gagattgata tcgaattttt
     3601 aggaaaagac acaacgaagg ttcaatttaa ctattataca aatggtgcag gaaaccatga
     3661 gaagattgtt gatctcgggt ttgatgcagc caatgcctat catacgtatg cattcgattg
     3721 gcagccaaac tctattaaat ggtatgtcga cgggcaatta aaacatactg caacaaacca
     3781 aattccgaca acacctggaa agatcatgat gaacttgtgg aatggcacgg gtgtcgatga
     3841 atggcttggc tcctacaatg gtgtaaatcc gctatacgct cattatgact gggtgcgcta
     3901 tacaaaaaaa taatgccaaa tgtgaaagag cctgctgcct cgaggggggg cccggtaccc
     3961 agcttttgtt ccctttagtg agggttaatt tcgagcttgg cgtaatcatg gtcatagctg
     4021 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4081 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4141 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4201 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4261 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4321 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4381 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4441 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4501 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4561 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4621 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4681 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4741 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4801 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     4861 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4921 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4981 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5041 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5101 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5161 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5221 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5281 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5341 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5401 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5461 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5521 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5581 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5641 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5701 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5761 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5821 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5881 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5941 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6001 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6061 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6121 caaatagggg ttccgcgcac atttccccga aaagtgc
//
