LOCUS       V005436                 7681 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005436
VERSION     V005436
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7681)
  AUTHORS   Fernandez-Martinez LT, Bibb MJ.
  TITLE     Use of the meganuclease I-SceI of Saccharomyces cerevisiae to select
            for gene deletions in actinomycetes
  JOURNAL   Sci Rep 4, 7100 (2014)
   PUBMED   25403842
REFERENCE   2  (bases 1 to 7681)
  AUTHORS   Fernandez-Martinez LT, Bibb MJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-JAN-2016) Biology, Edge Hill University, St Helens
            Road, Ormskirk L39 4QP, UK
REFERENCE   3  (bases 1 to 7681)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7681)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep 4,
            7100 (2014)"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (13-JAN-2016) Biology, Edge Hill University, St Helens Road,
            Ormskirk L39 4QP, UK"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            complement(467..576)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             892..1692
                     /label="ApmR"
                     /note="aminoglycoside 3-N-acetyltransferase type IV"
     rep_origin      1994..2582
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    2870..2891
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2906..2936
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    2944..2960
                     /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     2968..2984
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     regulatory      3021..3102
                     /gene="ermE*"
                     /regulatory_class="promoter"
     gene            3021..3102
                     /gene="ermE*"
                     /label="ermE*"
     CDS             3308..4000
                     /gene="SCEI"
                     /label="Intron-encoded endonuclease I-SceI"
                     /note="Intron-encoded endonuclease I-SceI from
                     Saccharomyces cerevisiae (strain ATCC 204508 / S288c).
                     Accession#: P03882"
     terminator      4007..4055
                     /label="fd terminator"
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     terminator      complement(4279..4327)
                     /label="fd terminator"
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             4671..5477
                     /gene="tsnR"
                     /label="23S rRNA (adenosine(1067)-2'-O)-methyltransferase"
                     /note="23S rRNA (adenosine(1067)-2'-O)-methyltransferase
                     from Streptomyces azureus. Accession#: P18644"
     CDS             6043..7482
                     /label="pSG5 Rep"
                     /note="replication initiator protein from the Streptomyces
                     ghanaensis plasmid pSG5"
ORIGIN
        1 ggccgtcgac gacgacgcgt ccgcctgcct gcttttggcc ggcgggccgg gccgcggttg
       61 tccgccgggt cggaccggtc ccttggtcga ccgatgcctc ggccacgacg atgctgggcc
      121 cgaagcggta cgcgaacccg tgccgggctg cggcggatcg cggcagcagg agctccccgc
      181 ccgccacggc tggctggatg gtcgcctcac gcgcccgcag gggtgacggc gtgggtgcgg
      241 cggtcagccc cgtgatcggg cgtcagacgg ccgctgaggg accgcgacca cccctccctt
      301 cccgcctgcc gtctttcacc gcccggccgg tccgcagacg ccgtacggcc aggccacgga
      361 ggattacggg cagatggcgc ccacccggca cccgggctgg ggcaccctgg acagcacgaa
      421 ggcccgcccc actcaccggg gcgggccttc gcgttgagct agagttcccg ccagcctcgc
      481 agagcaggat tcccgttgag caccgccagg tgcgaataag ggacagtgaa gaaggaacac
      541 ccgctcgcgg gtgggcctac ttcacctatc ctgcccggct gacgccgttg gatacaccaa
      601 ggaaagtcta cacgaaccct ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat
      661 accgaaaaaa tcgctataat gaccccgaag cagggttatg cagcggaaaa tgcagctcac
      721 ggtaactgat gccgtatttg cagtaccagc gtacggccca cagaatgatg tcacgctgaa
      781 aatgccggcc tttgaatggg ttcatgtgca gctccatcag caaaagggga tgataagttt
      841 atcaccaccg actatttgca acagtgccgt tgatcgtgct atgatcgact gatgtcatca
      901 gcggtggagt gcaatgtcgt gcaatacgaa tggcgaaaag ccgagctcat cggtcagctt
      961 ctcaaccttg gggttacccc cggcggtgtg ctgctggtcc acagctcctt ccgtagcgtc
     1021 cggcccctcg aagatgggcc acttggactg atcgaggccc tgcgtgctgc gctgggtccg
     1081 ggagggacgc tcgtcatgcc ctcgtggtca ggtctggacg acgagccgtt cgatcctgcc
     1141 acgtcgcccg ttacaccgga ccttggagtt gtctctgaca cattctggcg cctgccaaat
     1201 gtaaagcgca gcgcccatcc atttgccttt gcggcagcgg ggccacaggc agagcagatc
     1261 atctctgatc cattgcccct gccacctcac tcgcctgcaa gcccggtcgc ccgtgtccat
     1321 gaactcgatg ggcaggtact tctcctcggc gtgggacacg atgccaacac gacgctgcat
     1381 cttgccgagt tgatggcaaa ggttccctat ggggtgccga gacactgcac cattcttcag
     1441 gatggcaagt tggtacgcgt cgattatctc gagaatgacc actgctgtga gcgctttgcc
     1501 ttggcggaca ggtggctcaa ggagaagagc cttcagaagg aaggtccagt cggtcatgcc
     1561 tttgctcggt tgatccgctc ccgcgacatt gtggcgacag ccctgggtca actgggccga
     1621 gatccgttga tcttcctgca tccgccagag gcgggatgcg aagaatgcga tgccgctcgc
     1681 cagtcgattg gctgagctca taagttccta ttccgaagtt cctattcgaa atgaccgacc
     1741 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     1801 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     1861 tgctggagtt cttcgcccac cccaaaagga tctaggtgaa gatccttttt gataatctca
     1921 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     1981 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2041 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2101 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     2161 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2221 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2281 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2341 tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga gaaagcgcca
     2401 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2461 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2521 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2581 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     2641 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     2701 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     2761 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     2821 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     2881 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     2941 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgaattgt
     3001 acgtaggatc cagcccgacc cgagcacgcg ccggcacgcc tggtcgatgt cggaccggag
     3061 ttcgaggtac gcggcttgca ggtccaggaa ggggacgtcc atgcgagtgt ccgttcgagt
     3121 ggcggcttgc gcccgatgct agtcgcggtt gatcggcgat cgcaggtgca cgcggtcgat
     3181 cttgacggct ggcgagaggt gcggggagga tctgaccgac gcggtccaca cgtggcaccg
     3241 cgatgctgtt gtgggcacaa tcgtgccggt tggtaggatc gtctagaaca ggaggcccca
     3301 tatgcaccag aagaaccagg tcatgaacct gggcccgaac tccaagctgc tgaaggagta
     3361 caagtcccag ctgatcgagc tgaacatcga gcagttcgag gccggcatcg gcctgatcct
     3421 gggcgacgcc tacatccgct cccgcgacga gggcaagacc tactgcatgc agttcgagtg
     3481 gaagaacaag gcctacatgg accacgtctg cctgctgtac gaccagtggg tcctgtcccc
     3541 gccgcacaag aagcagcgcg tcaaccacct gggcaacctg gtcatcacct ggggcgccca
     3601 gaccttcaag caccaggcct tcaacaagct ggccaacctg ttcatcgtca acaacaagaa
     3661 gaccatcccg aacaacctgg tcgagaacta cctgaccccg atgtccctgg cctactggtt
     3721 catggacgac ggcggcaagt gggactacaa caagaactcc accaacaagt ccatcgtcct
     3781 gaacacccag tccttcacct tcgaggaggt cgagtacctg gtcaagggcc tgcgcaacaa
     3841 gttccagctg aactgctacg tcaagatcaa caagaacaag ccgatcatct acatcgactc
     3901 catgtcctac ctgatcttct acaacctgat caagccgtac ctgatcccgc agatgatgta
     3961 caagctgccg aacaccatct cctccgagac cttcctgaag tgataaaccg atacaattaa
     4021 aggctccttt tggagccttt ttttttggag attttcaacg tgaaaaaatt attattcgca
     4081 attccaacaa aaggctggag ctccaccgcg gccgcgcgcg atatcgaatt ccccagatct
     4141 aaagttttgt cgtctttcca gacgttagta aatgaatttt ctgtatgagg ttttgctaaa
     4201 caactttcaa cagtttcagc ggagtgagaa tagaaaggaa caactaaagg aattgcgaat
     4261 aataattttt tcacgttgaa aatctccaaa aaaaaaggct ccaaaaggag cctttaattg
     4321 tatcggttta tcagcttgct ttcgaggtga atttcttaaa cagcttgata ccgatagttg
     4381 cgccgacaat gacaacaacc atcgcccacg cataaccgat atattcggtc gctgaggctt
     4441 gcagggagtc aaaggccgct tttgcgggat ctcgtcgaag gcactagaag gcctatatgc
     4501 ggggatcgac cgcgcgggtc ccggacgggg aagagcgggg agctttgcca gagagcgacg
     4561 acttcccctt gcgttggtga ttgccggtca gggcagccat ccgccatcgt cgcgtagggt
     4621 gtcacacccc aggaatcgcg tcactgaaca cagcagccgg taggacgacc atgactgagt
     4681 tggacaccat cgcaaatccg tccgatcccg cggtgcagcg gatcatcgat gtcaccaagc
     4741 cgtcgcgatc caacataaag acaacgttga tcgaggacgt cgagcccctc atgcacagca
     4801 tcgcggccgg ggtggagttc atcgaggtct acggcagcga cagcagtcct tttccatctg
     4861 agttgctgga tctgtgcggg cggcagaaca taccggtccg cctcatcgac tcctcgatcg
     4921 tcaaccagtt gttcaagggg gagcggaagg ccaagacatt cggcatcgcc cgcgtccctc
     4981 gcccggccag gttcggcgat atcgcgagcc ggcgtgggga cgtcgtcgtt ctcgacgggg
     5041 tgaagatcgt cgggaacatc ggcgcgatag tacgcacgtc gctcgcgctc ggagcgtcgg
     5101 ggatcatcct ggtcgacagt gacatcacca gcatcgcgga ccggcgtctc caaagggcca
     5161 gccgaggtta cgtcttctcc cttcccgtcg ttctctccgg tcgcgaggag gccatcgcct
     5221 tcattcggga cagcggtatg cagctgatga cgctcaaggc ggatggcgac atttccgtga
     5281 aggaactcgg ggacaatccg gatcggctgg ccttgctgtt cggcagcgaa aagggtgggc
     5341 cttccgacct gttcgaggag gcgtcttccg cctcggtttc catccccatg atgagccaga
     5401 ccgagtctct caacgtttcc gtttccctcg gaatcgcgct gcacgagagg atcgacagga
     5461 atctcgcggc caaccgataa gcgcctctgt tcctcggacg ctcggttcct cgacctcgat
     5521 tcgtcagtga tgatcaacag gacccagaaa tggcacgagc ccgggattgg cctcccgggc
     5581 tcgccgcccc cgaaccggcg cagcatgccg agtgaggacg tttgctctgt gacgagcgta
     5641 cgagagaaca gcgcgtcgac accaccgcgc cccgccggac cgaagtgcca ctgcggcacc
     5701 ccgatcgtcc acatccccgg gaagcgcccc aaaatctact gctcggacgc ctgccgaaag
     5761 cgcgcaaagc gtctcattgc ggaccagtca cgcagcgtag caagcacgac tacagcgggt
     5821 aatccggacg cgggctttag tagtaaggga aaatctccgt ccactgcagg tcagagcgct
     5881 gtaactggaa cgggttctac cccaaacgag ggtgccgcga cagccgggcg ggaggacccg
     5941 cgcgacgagc ggttcgcccg gcgggaccgg caccagacgg tgtcgctgaa cgaggcgttc
     6001 aagggctgcg ggacccggct cactgccggg acggccgagc tgatgtggaa gccgggcgag
     6061 gcgacgtggg gcaacacgtg ccgctgcaac aacgtccaca cctgcccgtg gtgcatgagc
     6121 cgcatcctgg ccgtacgggg ctcgaacgtg cagctcgccg ccgacggtct cgcggacgcc
     6181 gggtacgggc tgcaccttgg cacgaacacg ttgcgccact tcgagcgcat ggcgttcggc
     6241 accgtccgca agggaatgcg gcacggcctg gtcgcggtct tgcacgacgg ctggaagggc
     6301 gcgtacggct cgtccggccg ccgctggcgc acgatgcgcg acgacttcgg gatcatcggc
     6361 tacgagcggg ccttcgagga caccttcggc tggggctccg gctggcacct gcactggcac
     6421 acgctctggg tcacccgcga ggtgctcggc ccggacgccc aggccgcgtt ccgcgacgca
     6481 ctcgccggag cctgggccgc cggggtcgaa tccgcgggcg gctacaccgt cagcgagacg
     6541 tgcgaccggc ccggttgctc ctgtgagggc aagggccacg gcaccgacgt gcgcccactc
     6601 aacggcgcgg acgccgctga cggcgatgcg ggcaagcagg cccgctacct ctacaaggac
     6661 ggcgacaaga ccaagggcgg cgtcgccaag atcggcctgg agctggccgg acagaacttc
     6721 aaggccggtc gcggcgacga ccgcatggga ccgctcgacc tgggcgacgc ggcggccgcc
     6781 gagctgcagc ggcttcgtcg gcccggcccg ttcgtcgaga agtaccgcga gcgcgagttc
     6841 ggcgtcttcc aggtccgcaa gcactaccgg tcgcagaacc tgaaccggct gatcaaggaa
     6901 ctcggcatcc agcaggacgt ccgcaccgag gaggagatca ccgacgacac cgagggcctg
     6961 gtcgccatcg cggtcatccc ggcctacatc tggtaccgat acatcgcccg cgtcgccgga
     7021 cgtcgcctcg acctcatcaa ggtcgccgag acgtacggcc taccgggcgt ccggcggctc
     7081 gtcgagtcct ggggcctggt gtggggcaag gacgtcctgg acccgcccgc cccggaagcg
     7141 ccggccgcgc cgggcgacct tgacgctgac cagatgaggt tcgaggtcat gtccgaggag
     7201 gaagctgctt tccgcgaggc gcgccggaag gccaacgagg cccgcacgga ggagctggcc
     7261 gcttcgctcg accgagtgcg tcagccgaag aaggaggcga tccgcccgac gatcagcctt
     7321 cggaagcgcc tgaagccgaa gccggtcacc gtcgacgtca agactccccc gcccggcgcc
     7381 gcgtcgcccg tgtgccgccg ctgcaagggc aagctcgccc cggtactcca gccctggggc
     7441 cggctccccg gagactgcct ccgcgtcgac acagcggtcg cgtagggccg ctgagggacc
     7501 gccatgagac cgccaccccg gacatccggg tggcggtctc atggcggtcc cggcgcgaaa
     7561 gccgagggtc tgcctgccgt gaggtggccg gcggacctgg gttactgctg gaatcccgcg
     7621 cccagtccga gcgtccgccg gcgcttttca cctggctccg ggtgctcgtc gatgagcacc
     7681 a
//
