LOCUS       40924_25446        3562 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pIJ12738, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3562)
  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 3562)
  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 3562)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3562)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep 4, 
            7100 (2014)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-JAN-2016) Biology, Edge Hill University, St Helens Road, 
            Ormskirk L39 4QP, UK"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     423..439
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     490..506
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(609..622)
                     /label=fragment of lacZ alpha peptide
                     /note="fragment of lacZ alpha peptide"
     primer_bind     complement(623..639)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(647..663)
                     /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(671..701)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(716..737)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1025..1613)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             1817..2617
                     /codon_start=1
                     /label=ApmR
                     /note="aminoglycoside 3-N-acetyltransferase type IV"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     oriT            3040..3149
                     /label=oriT
                     /note="incP origin of transfer"
ORIGIN
        1 atctacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
       61 gctctcgcag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
      121 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttg atcggcactt
      181 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattta tgcggtgtga
      241 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
      301 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      361 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      421 ttgtaaaacg acggccagtg ccaagcttgg gctgcaggtc gactctagag gatccgcggt
      481 ggcggccgct ctagaactag tggatccccc gggctgcagg aattcgatat cgagcttacc
      541 gtcgacctcg agggggggcc cggtacccaa ttcgccctat agtgagttag ggataacagg
      601 gtaatatcga attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca
      661 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
      721 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
      781 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
      841 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
      901 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
      961 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1021 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1081 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1141 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1201 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     1261 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1321 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1381 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1441 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1501 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1561 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1621 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     1681 tggtcatgag attatcaaaa aggatcttca cctagatcct tttggttcat gtgcagctcc
     1741 atcagcaaaa ggggatgata agtttatcac caccgactat ttgcaacagt gccgttgatc
     1801 gtgctatgat cgactgatgt catcagcggt ggagtgcaat gtcgtgcaat acgaatggcg
     1861 aaaagccgag ctcatcggtc agcttctcaa ccttggggtt acccccggcg gtgtgctgct
     1921 ggtccacagc tccttccgta gcgtccggcc cctcgaagat gggccacttg gactgatcga
     1981 ggccctgcgt gctgcgctgg gtccgggagg gacgctcgtc atgccctcgt ggtcaggtct
     2041 ggacgacgag ccgttcgatc ctgccacgtc gcccgttaca ccggaccttg gagttgtctc
     2101 tgacacattc tggcgcctgc caaatgtaaa gcgcagcgcc catccatttg cctttgcggc
     2161 agcggggcca caggcagagc agatcatctc tgatccattg cccctgccac ctcactcgcc
     2221 tgcaagcccg gtcgcccgtg tccatgaact cgatgggcag gtacttctcc tcggcgtggg
     2281 acacgatgcc aacacgacgc tgcatcttgc cgagttgatg gcaaaggttc cctatggggt
     2341 gccgagacac tgcaccattc ttcaggatgg caagttggta cgcgtcgatt atctcgagaa
     2401 tgaccactgc tgtgagcgct ttgccttggc ggacaggtgg ctcaaggaga agagccttca
     2461 gaaggaaggt ccagtcggtc atgcctttgc tcggttgatc cgctcccgcg acattgtggc
     2521 gacagccctg ggtcaactgg gccgagatcc gttgatcttc ctgcatccgc cagaggcggg
     2581 atgcgaagaa tgcgatgccg ctcgccagtc gattggctga gctcatgagc ggagaacgag
     2641 atgacgttgg aggggcaagg tcgcgctgat tgctggggca acacgtggag cggatcgggg
     2701 attgtctttc ttcagctcgc tgatgatatg ctgacgctca atgccgtttg gcctccgact
     2761 aacgaaaatc ccgcatttgg acggctgatc cgattggcac ggcggacggc gaatggcgga
     2821 gcagacgctc gtccgggggc aatgagatat gaaaaagcct gaactcaccg cgacgtatcg
     2881 atgnnnntat gcaggtcgac ggatcttttc cgctgcataa ccctgcttcg gggtcattat
     2941 agcgattttt tcggtatatc catccttttt cgcacgatat acaggatttt gccaaagggt
     3001 tcgtgtagac tttccttggt gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg
     3061 cccacccgcg agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa
     3121 cgggaatcct gctctgcgag gctggccggc atgagcctgt cggcctacct gctggccgtc
     3181 ggccagggct acaaaatcac gggcgtcgtg gactatgagc acgtccgcga gctggcccgc
     3241 atcaatggcg acctgggccg cctgggcggc ctgctgaaac tctggctcac cgacgacccg
     3301 cgcacggcgc ggttcggtga tgccacgatc ctcgccctgc tggcgaagat cgaagagaag
     3361 caggacgagc ttggcaaggt catgatgggc gtggtccgcc cgagggcaga gccatgactt
     3421 ttttagccgc taaaacggcc ggggggtgcg cgtgattgcc aagcacgtcc ccatgcgctc
     3481 catcaagaag agcgacttcg cggagctggt gaagtacatc accgacgagc aaggcaagac
     3541 cgatccccgg ggacctgcat ag
//
