LOCUS       40924_7326        4799 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBSc243B, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4799)
  AUTHORS   Radeck J, Meyer D, Lautenschlager N, Mascher T.
  TITLE     Bacillus SEVA siblings: A Golden Gate-based toolbox to create 
            personalized integrative vectors for Bacillus subtilis
  JOURNAL   Sci Rep 7 (1), 14134 (2017)
  PUBMED    29074996
REFERENCE   2  (bases 1 to 4799)
  AUTHORS   Radeck J.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-APR-2017) Biology, Technische Universitat Dresden, 
            Zellescher Weg 20b, Dresden 01627, Germany
REFERENCE   3  (bases 1 to 4799)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4799)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep"; 
            date: "2017"; volume: "7"; issue: "1"; pages: "14134"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-APR-2017) Biology, Technische Universitat Dresden, Zellescher 
            Weg 20b, Dresden 01627, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4799
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    125..146
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        161..191
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    199..215
                     /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     223..239
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    249..305
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(309..325)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      604..698
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             877..1689
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            1859..1967
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      2061..2649
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2706..3536)
                     /codon_start=1
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
                     /translation="VASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     rep_origin      3550..3901
                     /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)"
     CDS             complement(4060..4464)
                     /codon_start=1
                     /gene="bleO"
                     /product="BleO"
                     /label=bleO
                     /protein_id="ATU32889.1"
                     /translation="MRMLQSIPALPVGDIKKSIGFYCDKLGFTLVHHEDGFAVLMCNEV
                     RIHLWEASDEGWRSRSNDSPVCTGAESFIAGTASCRIEVEGIDELYQHIKPLGILHPNT
                     SLKDQWWDERDFAVIDPDNNLISFFQQIKS"
     gene            complement(4060..4464)
                     /gene="bleO"
                     /label=bleO
     terminator      complement(4707..4793)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 ttaattaaga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
       61 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
      121 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
      181 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
      241 atgattacga attcgagctc ggtacccggg gatcctctag agtcgacctg caggcatgca
      301 agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
      361 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
      421 accgatcgcc cttcctaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
      481 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
      541 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgaactagtc
      601 ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa
      661 cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aggggtcccc aataattacg
      721 atttgagatg agaccttgca ggtgagatga gacggtcttc aacatcgcag taaatttgtg
      781 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
      841 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca gcgtgaaacg
      901 agctgtagcc gtccgcgtct gaacagcaac atggatgcgg atctgtatgg ctataaatgg
      961 gcgcgtgata acgtgggtca gagcggcgcg accatttatc gtctgtatgg caaaccggat
     1021 gcgccggaac tgtttctgaa acatggcaaa ggcagcgtgg cgaacgatgt gaccgatgaa
     1081 atggtgcgtc tgaactggct gaccgaattt atgccgctgc cgaccattaa acattttatt
     1141 cgcaccccgg atgatgcgtg gctgctgacc accgcgattc cgggcaaaac cgcgtttcag
     1201 gtgctggaag aatatccgga tagcggcgaa aacattgtgg atgcgctggc cgtgtttctg
     1261 cgtcgtctgc atagcattcc ggtgtgcaac tgcccgttta acagcgatcg tgtgtttcgt
     1321 ctggcccagg cgcagagccg tatgaacaac ggcctggtgg atgcgagcga ttttgatgat
     1381 gaacgtaacg gctggccggt ggaacaggtg tggaaagaaa tgcataaact gctgccgttt
     1441 agcccggata gcgtggtgac ccacggcgat tttagcctgg ataacctgat tttcgatgaa
     1501 ggcaaactga ttggctgcat tgatgtgggc cgtgtgggca ttgcggatcg ttatcaggat
     1561 ctggccattc tgtggaactg cctgggcgaa tttagcccga gcctgcaaaa acgtctgttt
     1621 cagaaatatg gcattgataa tccggatatg aacaaactgc aatttcatct gatgctggat
     1681 gaatttttct aataattaat tggaccgcgg tccgcgcgtt gtccttttcc gctgcataac
     1741 cctgcttcgg ggtcattata gcgatttttt cggtatatcc atcctttttc gcacgatata
     1801 caggattttg ccaaagggtt cgtgtagact ttccttggtg tatccaacgg cgtcagccgg
     1861 gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc cttcttcact gtcccttatt
     1921 cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg ctggccgtag gccggccgat
     1981 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     2041 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2101 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2161 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     2221 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2281 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2341 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2401 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2461 agcgccacgc ttcccgaagg gagaaaggcg gacaggcatc cggtaagcgg cagggtcgga
     2521 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     2581 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     2641 ctatggaaaa acgccagcaa cgcggccgtg aaaggcaggc cggtccgtgg tggccacggc
     2701 ctctaggcca gatccagcgg catctgggtt agtcgagcgc gggccgcttc ccatgtctca
     2761 ccagggcgag cctgtttcgc gatctcagca tctgaaatct tcccggcctt gcgcttcgct
     2821 ggggccttac ccaccgcctt ggcgggcttc ttcggtccaa aactgaacaa cagatgtgtg
     2881 accttgcgcc cggtctttcg ctgcgcccac tccacctgta gcgggctgtg ctcgttgatc
     2941 tgcgtcacgg ctggatcaag cactcgcaac ttgaagtcct tgatcgaggg ataccggcct
     3001 tccagttgaa accactttcg cagctggtca atttctattt cgcgctggcc gatgctgtcc
     3061 cattgcatga gcagctcgta aagcctgatc gcgtgggtgc tgtccatctt ggccacgtca
     3121 gccaaggcgt atttggtgaa ctgtttggtg agttccgtca ggtacggcag catgtctttg
     3181 gtgaacctga gttctacacg gccctcaccc tcccggtaga tgattgtttg cacccagccg
     3241 gtaatcatca cactcggtct tttccccttg ccattgggct cttgggttaa ccggacttcc
     3301 cgccgtttca ggcgcagggc cgcttctttg agctggttgt aggaagattc gatagggaca
     3361 cccgccatcg tcgctatgtc ctccgccgtc actgaataca tcacttcatc ggtgacaggc
     3421 tcgctcctct tcacctggct aatacaggcc agaacgatcc gctgttcctg aacactgagg
     3481 cgatacgcgg cctcgaccag ggcattgctt ttgtaaacca ttgggggtga ggccacgttc
     3541 gacattcctt gtgtataagg ggacactgta tctgcgtccc acaatacaac aaatccgtcc
     3601 ctttacaaca acaaatccgt cccttcttaa caacaaatcc gtcccttaat ggcaacaaat
     3661 ccgtcccttt ttaaactcta caggccacgg attacgtggc ctgtagacgt cctaaaaggt
     3721 ttaaaaggga aaaggaagaa aagggtggaa acgcaaaaaa cgcaccacta cgtggccccg
     3781 ttggggccgc atttgtgccc ctgaaggggc gggggaggcg tctgggcaat ccccgtttta
     3841 ccagtcccct atcgccgcct gagagggcgc aggaagcgag taatcagggt atcgaggcgg
     3901 attcaccctt ggcgtccaac cagcggcacc agcggcgcct gagaggggcg cgttcacctg
     3961 cgatggcgaa gacttgcgag gtctcagcga tcgtctcagc gaggcgcgtt cttttattca
     4021 gcaatcgcgc ccgattgctg aacagattaa taatagattt tagcttttta tttgttgaaa
     4081 aaagctaatc aaattgttgt cgggatcaat tactgcaaag tctcgttcat cccaccactg
     4141 atcttttaat gatgtattgg ggtgcaaaat gcccaaaggc ttaatatgtt gatataattc
     4201 atcaattccc tctacttcaa tgcggcaact agcagtacca gcaataaacg actccgcacc
     4261 tgtacaaacc ggtgaatcat tactacgaga gcgccagcct tcatcacttg cctcccatag
     4321 atgaatccga acctcattac acattagaac tgcgaatcca tcttcatggt gaaccaaagt
     4381 gaaacctagt ttatcgcaat aaaaacctat actcttttta atatccccga ctggcaatgc
     4441 cgggatagac tgtaacattc tcacgcataa aatccccttt cattttctaa tgtaaatcta
     4501 ttaccttatt attaattcaa ttcgctcata attaatcctt tttcttatta cgcaaaatgg
     4561 cccgatttaa gcacaccctt tattccgtta atgcgccatg acagccatga taattactaa
     4621 tactaggaga agttaataaa tacgtaacca acatgattaa caattattag aggtcatcgt
     4681 acgcgcccag ctgtctaggg cggcggattt gtcctactca ggagagcgtt caccgacaaa
     4741 caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcct
//
