LOCUS       V005437                 7744 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005437
VERSION     V005437
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7744)
  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 7744)
  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 7744)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7744)
  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..7744
                     /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"
     terminator      complement(3163..3197)
                     /label="lambda t0 terminator"
                     /note="minimal transcription terminator from phage lambda
                     (Scholtissek and Grosse, 1987)"
     regulatory      3232..3332
                     /gene="tipA"
                     /regulatory_class="promoter"
     gene            3232..3332
                     /gene="tipA"
                     /label="tipA"
     CDS             3371..4063
                     /gene="SCEI"
                     /label="Intron-encoded endonuclease I-SceI"
                     /note="Intron-encoded endonuclease I-SceI from
                     Saccharomyces cerevisiae (strain ATCC 204508 / S288c).
                     Accession#: P03882"
     terminator      4070..4118
                     /label="fd terminator"
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     terminator      complement(4342..4390)
                     /label="fd terminator"
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             4734..5540
                     /gene="tsnR"
                     /label="23S rRNA (adenosine(1067)-2'-O)-methyltransferase"
                     /note="23S rRNA (adenosine(1067)-2'-O)-methyltransferase
                     from Streptomyces azureus. Accession#: P18644"
     CDS             6106..7545
                     /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 acgtaggcgt cagctacgat gttccgggga ctgctgatcc ggtcagcagg tggaagaggg
     3061 actggattcc aaagttctca atgctgcttg ctgttcttga atggggggtc gttgacgacg
     3121 acatggctcg attggcgcga caagttgctg cgattctcac caataaaaaa cgcccggcgg
     3181 caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg accggatcgg
     3241 ggatctgggc tgagggagcc gacggcacgc ggcggctcac ggcgtggcac gcggaacgtc
     3301 cgggcttgca cctcacgtca cgtgaggagg cagcgtggac ggcgtcagag aagggagcgg
     3361 acatatgcac cagaagaacc aggtcatgaa cctgggcccg aactccaagc tgctgaagga
     3421 gtacaagtcc cagctgatcg agctgaacat cgagcagttc gaggccggca tcggcctgat
     3481 cctgggcgac gcctacatcc gctcccgcga cgagggcaag acctactgca tgcagttcga
     3541 gtggaagaac aaggcctaca tggaccacgt ctgcctgctg tacgaccagt gggtcctgtc
     3601 cccgccgcac aagaagcagc gcgtcaacca cctgggcaac ctggtcatca cctggggcgc
     3661 ccagaccttc aagcaccagg ccttcaacaa gctggccaac ctgttcatcg tcaacaacaa
     3721 gaagaccatc ccgaacaacc tggtcgagaa ctacctgacc ccgatgtccc tggcctactg
     3781 gttcatggac gacggcggca agtgggacta caacaagaac tccaccaaca agtccatcgt
     3841 cctgaacacc cagtccttca ccttcgagga ggtcgagtac ctggtcaagg gcctgcgcaa
     3901 caagttccag ctgaactgct acgtcaagat caacaagaac aagccgatca tctacatcga
     3961 ctccatgtcc tacctgatct tctacaacct gatcaagccg tacctgatcc cgcagatgat
     4021 gtacaagctg ccgaacacca tctcctccga gaccttcctg aagtgataaa ccgatacaat
     4081 taaaggctcc ttttggagcc tttttttttg gagattttca acgtgaaaaa attattattc
     4141 gcaattccaa caaaaggctg gagctccacc gcggccgcgc gcgatatcga attccccaga
     4201 tctaaagttt tgtcgtcttt ccagacgtta gtaaatgaat tttctgtatg aggttttgct
     4261 aaacaacttt caacagtttc agcggagtga gaatagaaag gaacaactaa aggaattgcg
     4321 aataataatt ttttcacgtt gaaaatctcc aaaaaaaaag gctccaaaag gagcctttaa
     4381 ttgtatcggt ttatcagctt gctttcgagg tgaatttctt aaacagcttg ataccgatag
     4441 ttgcgccgac aatgacaaca accatcgccc acgcataacc gatatattcg gtcgctgagg
     4501 cttgcaggga gtcaaaggcc gcttttgcgg gatctcgtcg aaggcactag aaggcctata
     4561 tgcggggatc gaccgcgcgg gtcccggacg gggaagagcg gggagctttg ccagagagcg
     4621 acgacttccc cttgcgttgg tgattgccgg tcagggcagc catccgccat cgtcgcgtag
     4681 ggtgtcacac cccaggaatc gcgtcactga acacagcagc cggtaggacg accatgactg
     4741 agttggacac catcgcaaat ccgtccgatc ccgcggtgca gcggatcatc gatgtcacca
     4801 agccgtcgcg atccaacata aagacaacgt tgatcgagga cgtcgagccc ctcatgcaca
     4861 gcatcgcggc cggggtggag ttcatcgagg tctacggcag cgacagcagt ccttttccat
     4921 ctgagttgct ggatctgtgc gggcggcaga acataccggt ccgcctcatc gactcctcga
     4981 tcgtcaacca gttgttcaag ggggagcgga aggccaagac attcggcatc gcccgcgtcc
     5041 ctcgcccggc caggttcggc gatatcgcga gccggcgtgg ggacgtcgtc gttctcgacg
     5101 gggtgaagat cgtcgggaac atcggcgcga tagtacgcac gtcgctcgcg ctcggagcgt
     5161 cggggatcat cctggtcgac agtgacatca ccagcatcgc ggaccggcgt ctccaaaggg
     5221 ccagccgagg ttacgtcttc tcccttcccg tcgttctctc cggtcgcgag gaggccatcg
     5281 ccttcattcg ggacagcggt atgcagctga tgacgctcaa ggcggatggc gacatttccg
     5341 tgaaggaact cggggacaat ccggatcggc tggccttgct gttcggcagc gaaaagggtg
     5401 ggccttccga cctgttcgag gaggcgtctt ccgcctcggt ttccatcccc atgatgagcc
     5461 agaccgagtc tctcaacgtt tccgtttccc tcggaatcgc gctgcacgag aggatcgaca
     5521 ggaatctcgc ggccaaccga taagcgcctc tgttcctcgg acgctcggtt cctcgacctc
     5581 gattcgtcag tgatgatcaa caggacccag aaatggcacg agcccgggat tggcctcccg
     5641 ggctcgccgc ccccgaaccg gcgcagcatg ccgagtgagg acgtttgctc tgtgacgagc
     5701 gtacgagaga acagcgcgtc gacaccaccg cgccccgccg gaccgaagtg ccactgcggc
     5761 accccgatcg tccacatccc cgggaagcgc cccaaaatct actgctcgga cgcctgccga
     5821 aagcgcgcaa agcgtctcat tgcggaccag tcacgcagcg tagcaagcac gactacagcg
     5881 ggtaatccgg acgcgggctt tagtagtaag ggaaaatctc cgtccactgc aggtcagagc
     5941 gctgtaactg gaacgggttc taccccaaac gagggtgccg cgacagccgg gcgggaggac
     6001 ccgcgcgacg agcggttcgc ccggcgggac cggcaccaga cggtgtcgct gaacgaggcg
     6061 ttcaagggct gcgggacccg gctcactgcc gggacggccg agctgatgtg gaagccgggc
     6121 gaggcgacgt ggggcaacac gtgccgctgc aacaacgtcc acacctgccc gtggtgcatg
     6181 agccgcatcc tggccgtacg gggctcgaac gtgcagctcg ccgccgacgg tctcgcggac
     6241 gccgggtacg ggctgcacct tggcacgaac acgttgcgcc acttcgagcg catggcgttc
     6301 ggcaccgtcc gcaagggaat gcggcacggc ctggtcgcgg tcttgcacga cggctggaag
     6361 ggcgcgtacg gctcgtccgg ccgccgctgg cgcacgatgc gcgacgactt cgggatcatc
     6421 ggctacgagc gggccttcga ggacaccttc ggctggggct ccggctggca cctgcactgg
     6481 cacacgctct gggtcacccg cgaggtgctc ggcccggacg cccaggccgc gttccgcgac
     6541 gcactcgccg gagcctgggc cgccggggtc gaatccgcgg gcggctacac cgtcagcgag
     6601 acgtgcgacc ggcccggttg ctcctgtgag ggcaagggcc acggcaccga cgtgcgccca
     6661 ctcaacggcg cggacgccgc tgacggcgat gcgggcaagc aggcccgcta cctctacaag
     6721 gacggcgaca agaccaaggg cggcgtcgcc aagatcggcc tggagctggc cggacagaac
     6781 ttcaaggccg gtcgcggcga cgaccgcatg ggaccgctcg acctgggcga cgcggcggcc
     6841 gccgagctgc agcggcttcg tcggcccggc ccgttcgtcg agaagtaccg cgagcgcgag
     6901 ttcggcgtct tccaggtccg caagcactac cggtcgcaga acctgaaccg gctgatcaag
     6961 gaactcggca tccagcagga cgtccgcacc gaggaggaga tcaccgacga caccgagggc
     7021 ctggtcgcca tcgcggtcat cccggcctac atctggtacc gatacatcgc ccgcgtcgcc
     7081 ggacgtcgcc tcgacctcat caaggtcgcc gagacgtacg gcctaccggg cgtccggcgg
     7141 ctcgtcgagt cctggggcct ggtgtggggc aaggacgtcc tggacccgcc cgccccggaa
     7201 gcgccggccg cgccgggcga ccttgacgct gaccagatga ggttcgaggt catgtccgag
     7261 gaggaagctg ctttccgcga ggcgcgccgg aaggccaacg aggcccgcac ggaggagctg
     7321 gccgcttcgc tcgaccgagt gcgtcagccg aagaaggagg cgatccgccc gacgatcagc
     7381 cttcggaagc gcctgaagcc gaagccggtc accgtcgacg tcaagactcc cccgcccggc
     7441 gccgcgtcgc ccgtgtgccg ccgctgcaag ggcaagctcg ccccggtact ccagccctgg
     7501 ggccggctcc ccggagactg cctccgcgtc gacacagcgg tcgcgtaggg ccgctgaggg
     7561 accgccatga gaccgccacc ccggacatcc gggtggcggt ctcatggcgg tcccggcgcg
     7621 aaagccgagg gtctgcctgc cgtgaggtgg ccggcggacc tgggttactg ctggaatccc
     7681 gcgcccagtc cgagcgtccg ccggcgcttt tcacctggct ccgggtgctc gtcgatgagc
     7741 acca
//
