LOCUS       40924_27524        7554 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Over-expression shuttle vector for gene cloning and transfer from E.
            coli to Streptomyces. Contains inducible PnitA-NitR system .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7554)
  AUTHORS   Sun N,  Wang Z-B, Wu H-P, Mao X-M, Li Y-Q
  TITLE     Construction of over-expression shuttle vectors in Streptomyces
  JOURNAL   Annals of Microbiology Dec 2012, Volume 62, Issue 4, pp 1541-1546
REFERENCE   2  (bases 1 to 7554)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7554)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Annals of
            Microbiology Dec 2012, Volume 62, Issue 4, pp 1541-1546"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      55..94
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     terminator      1177..1216
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     primer_bind     complement(4297..4313)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4321..4337)
                     /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(4345..4375)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4390..4411)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(4528..4545)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4699..5287)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             5491..6291
                     /codon_start=1
                     /label=ApmR
                     /note="aminoglycoside 3-N-acetyltransferase type IV"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     CDS             complement(6728..7096)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(7129..7238)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     terminator      7402..7441
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
ORIGIN
        1 catatgctgg acggcggatc cgccgatatc gccaagcttc agagatctta aaccgataca
       61 attaaaggct ccttttggag cctttttttt tggaggcgaa ctcccttatg cgggtggcgc
      121 agaatgccag gacccttgtc attccacgtc aattcatgcg ccttttcacc tcgtactgtc
      181 ctgccaaaca caagcaacgg aggtacggac atgaacacct tcttctcctc cgaccaggtc
      241 tcggcgcccg accgcgtcgc gctctggcac gacgtcatct gccgcagcta cgtcccgctg
      301 aacatcaccc tgaccagcga gcagcccttc atcggcaccg tctccaccgg caacctgggc
      361 accgtccgca tcgccacgtc ctcgtccctg ccccagcaga tcacccgcac ccgccgcctg
      421 atccgccagg acgagcgcga gtacctcatg gtcggcgtgc agtccgccgg ccacgccctc
      481 gtgcagcagc acggccgcac cgcccgcgtc ggccgcggcg gcctggtctt ctgggacacc
      541 cgccacccgt acgacatcct cttcccgacc gactggcgca tgagcgtctt ccagttcccg
      601 cgctactcct tcggcttcac cgaggacttc atcggccgca tgaccgccgt caacgtcggc
      661 ggcgaccgcg gcatcggccg cgtggtctcc tccttcatga ccagcatcaa cgacgcgacc
      721 gacgccggcg acctggcgga ggtcgcctcc ctccacaaca gcgccgtcga cctgctgtcc
      781 gcggcgatcc ggaccgagct ggccgaccag gccgccgcct ccgacggcct gctcgagtgc
      841 gtgctcgcct acatccgcca gaacctggcc gacccgaacc tgtgcgcctc ccagatcgcc
      901 gccgagcaca acgtctccgt gcggaccctc caccgcctgt tctcggccac cggccagggc
      961 gtggccgagc acatccgcaa cctccgcctc gagcgcatca agaccgagct ggccgacccg
     1021 accagccggc gctacaccat cagcgccctg gcgcgcaagt ggggcttcct cgacccgtcc
     1081 accttctccc gcgcgttcaa ggacgcctac ggcatcaccg cccgcgagtg ggcggcctcc
     1141 gcctccgcct ccccgaccga ggtctcgtag taaaccgata caattaaagg ctccttttgg
     1201 agcctttttt tttggagtct agactgcagc cgggggtccg gcagggccgg agcgggcgga
     1261 accgtgctct gacctgcggc ccgagtttcg tcacgtgacg gaatggaagg ctgctgcatt
     1321 tcgtcacgtg acgtatctcg gcgagcgact gccgacgcca cggcggacac gatcgcctcg
     1381 cgctggcgcc gggcctcgta cgcccgctgg cggcaggagc ggcggcagta gtcccggctc
     1441 cggccgacgc cggattgctt gatctccgag ccgcaccagg cgcagagctt cgcgccgtcg
     1501 gcgtccctgg gggtggtggt gctcatggcc gacgaccgta cgcggcacgt ctcgtagcga
     1561 ggcgagtcgg gcgcgaggta ccgcctgcac gaagtgccgg cggggccgac cccgggcgag
     1621 taatcccagg attactcccg cggcttcgac cccggccgcc gtcgccgcgt acgtcaccga
     1681 cccccgccgt acgtcaccgg gatgacgtac ggcggggggg agcgagttag tgcgaagtgg
     1741 gcccacttgc gagccgggcg atgtgccggg cggcccgctc ctggcggtcg tcggcgtcgt
     1801 cgtcctggtc gtcgtcctgc tctcgccgtc ggcgtgcagt tgcttcctcg cggcgctggg
     1861 cgagggcggc gagcatgtcg gcgtacgcct cggccacctc ccccgccgtg agcaccacca
     1921 ctgtgtcggc cgcgtcggcc agcgccagga cctcccgcac ccgttcgccc acggccgccg
     1981 aatcctcgtt gccgtccttg ccttcggcgg cccgggtcgc ctcgaggtcg agggcgcggc
     2041 gggtgaccgc gtgccatccg tcctcggtca cggcgacccc ggcccgcagc tccccgccgt
     2101 cggcgtcggc cgccaggagc agatcgaggt cgtcggcctc ggtgtcgccg ccgtcgagcc
     2161 cgagcatctg ccgcaggtag cgggtccatt cgatggcccg gcgtccccgg gttgcccgct
     2221 cgtactcgtg ccagcgcgag aggttccact ccagcgagcc gaccccggcg gcgtcgtcct
     2281 cggtcatgcc gccggtcagg tccccgatcc gtccgaggag ttcgaacggg gcgacgttcc
     2341 cgccggtcgc cgtcttgagg tcggcgcggg cgagttcgag ggcgggcgcc ttcccgtcct
     2401 gggtcttggc gatgtactcg gcgaggtcgt tggcgtcgcg ctcggtctcc agccgcttga
     2461 agtcgacgcc gtgccggtcg tcgggcgtga aggcggggtt gaccttgcgc agggcggcgg
     2521 tccacacgga ccgccagtgc ccctgccact cgtcgagcgc ggcgccggtc ggctcgaagg
     2581 tggcgacgat ctgcttcgcg gaccgctccc cctcggtccg gccgccgacc aggacgatcg
     2641 cgtggatgtg cgggtgccag ccgttgatct gccccacggt gacttcggtc gcgcggatca
     2701 tgccgacgta cccgatccgg tctcggatgc cctcgcggtc ggcggcccgg tgcccgtcct
     2761 tggcccggcg tccggcccac gtgccgcccg tgatcagtcg ctggtaggcg cccggccgcc
     2821 gggggctgtc cggcgtcttc cgggtgccct ggagggcgtc catgaggtcc gcgagccggt
     2881 ccgtgtgccc atggcgggcc gtgaaggtga ccaggtaggc ggtccccccg cgcttgatcc
     2941 actcgaccac ggcggcggtg atctcctcgg cccgcttgtg ccggatcgtg gcggcgcaga
     3001 ccgggcagag ccagatccgc ccgcaccgca tcaggcccag gaccacggac gttccggccg
     3061 ccgtctgggc gacgattacg ccggaggcag ggtccatcag ggcgcggccg cagcccttgc
     3121 acgcggcgtc cccgctgatc cgccacagcg tccggcggcg gctgtaccgg gcggctttcc
     3181 gcagtcgggc agcgtcgctc cgcgacgtgc ttcctacttc cgagaggctg tcgcctctcg
     3241 ggctctcccc atccaccccg tccggagaaa ccgcaggtcg gaggggtgcg ggaaactctg
     3301 ttgtttcttt cccaaggtgt tcgcttttgc ctcgggcggc atctcgcgtc acacgcgcga
     3361 tcgcccgctt cgctgccatc cggcagcggt ctgagcagta gatacgcggc cgtttgcccg
     3421 gtgtgtgggc aattgcggtc ccgcagtggc agcggggccc ggcgggccga tctggcaatg
     3481 cctcggcatc gctccgtact ctgggcacga gcaacgttcc tgtctcgccc ggctaagggg
     3541 cgcgagtctg ggagcggacg ggtcggaggt gcgaagtccg gcccgttgct ctttggtctg
     3601 gtgggaatcc tggcaccaat cgggccagag gttccctccg ccactcccga cgccccttgg
     3661 ggctggtgtg acttggaggg ccgaagagag ccccgccggt gatccggcgg ggctttgacg
     3721 tgcggtcagt gcgtgtgtcg gcgagcgatg gccacgaggc cctggaagcc gagcggtccg
     3781 gcgaagtcgg cccagtcgca accgggctca gcgcagtggg cggaccagcc accgccgttg
     3841 gggtcctgga ccaggttcac ggtcccctcg gtcaggcgtc cgtcgaagtc ggtcatggtc
     3901 ggtctcctgg tgggtggggg cggggcgcca gcacgaagtg ccggcgcccc gcgggggttg
     3961 gtcgggtcag gcgccgaacc ggcgggcggc ggcggcgacc aggccgtcgg cggcggccat
     4021 ggcgatgatc cgctgcaccg cgggatcgga cggatttgcg atggtgtcca actcagtcat
     4081 ggtcgtccta ccggctgctg tgttcagtga cgcgattcct ggggtgtgac accctacgcg
     4141 acgatggcgg atggctgccc tgaccggcaa tcaccaacgc aaggggaagt cgtcgctctc
     4201 tggcaaagct ccccgctctt ccccgtccgg gacccgcgcg gtcgatcccc gcatatatga
     4261 agtattcgcc ttgatcccca tcgaattcgt aatcatgtca tagctgtttc ctgtgtgaaa
     4321 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4381 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4441 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4501 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4561 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4621 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4681 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4741 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4801 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4861 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4921 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4981 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5041 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5101 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5161 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5221 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5281 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5341 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttggt
     5401 tcatgtgcag ctccatcagc aaaaggggat gataagttta tcaccaccga ctatttgcaa
     5461 cagtgccgtt gatcgtgcta tgatcgactg atgtcatcag cggtggagtg caatgtcgtg
     5521 caatacgaat ggcgaaaagc cgagctcatc ggtcagcttc tcaaccttgg ggttaccccc
     5581 ggcggtgtgc tgctggtcca cagctccttc cgtagcgtcc ggcccctcga agatgggcca
     5641 cttggactga tcgaggccct gcgtgctgcg ctgggtccgg gagggacgct cgtcatgccc
     5701 tcgtggtcag gtctggacga cgagccgttc gatcctgcca cgtcgcccgt tacaccggac
     5761 cttggagttg tctctgacac attctggcgc ctgccaaatg taaagcgcag cgcccatcca
     5821 tttgcctttg cggcagcggg gccacaggca gagcagatca tctctgatcc attgcccctg
     5881 ccacctcact cgcctgcaag cccggtcgcc cgtgtccatg aactcgatgg gcaggtactt
     5941 ctcctcggcg tgggacacga tgccaacacg acgctgcatc ttgccgagtt gatggcaaag
     6001 gttccctatg gggtgccgag acactgcacc attcttcagg atggcaagtt ggtacgcgtc
     6061 gattatctcg agaatgacca ctgctgtgag cgctttgcct tggcggacag gtggctcaag
     6121 gagaagagcc ttcagaagga aggtccagtc ggtcatgcct ttgctcggtt gatccgctcc
     6181 cgcgacattg tggcgacagc cctgggtcaa ctgggccgag atccgttgat cttcctgcat
     6241 ccgccagagg cgggatgcga agaatgcgat gccgctcgcc agtcgattgg ctgagctcat
     6301 gagcggagaa cgagatgacg ttggaggggc aaggtcgcgc tgattgctgg ggcaacacgt
     6361 ggagcggatc ggggattgtc tttcttcagc tcgctgatga tatgctgacg ctcaatgccg
     6421 tttggcctcc gactaacgaa aatcccgcat ttggacggct gatccgattg gcacggcgga
     6481 cggcgaatgg cggagcagac gctcgtccgg gggcaatgag atatgaaaaa gcctgaactc
     6541 accgcgacgt atcgggccct ggccagctag ctagagtcga cctgcaggtc cccggggatc
     6601 ggtcttgcct tgctcgtcgg tgatgtactt caccagctcc gcgaagtcgc tcttcttgat
     6661 ggagcgcatg gggacgtgct tggcaatcac gcgcaccccc cggccgtttt agcggctaaa
     6721 aaagtcatgg ctctgccctc gggcggacca cgcccatcat gaccttgcca agctcgtcct
     6781 gcttctcttc gatcttcgcc agcagggcga ggatcgtggc atcaccgaac cgcgccgtgc
     6841 gcgggtcgtc ggtgagccag agtttcagca ggccgcccag gcggcccagg tcgccattga
     6901 tgcgggccag ctcgcggacg tgctcatagt ccacgacgcc cgtgattttg tagccctggc
     6961 cgacggccag caggtaggcc gacaggctca tgccggccgc cgccgccttt tcctcaatcg
     7021 ctcttcgttc gtctggaagg cagtacacct tgataggtgg gctgcccttc ctggttggct
     7081 tggtttcatc agccatccgc ttgccctcat ctgttacgcc ggcggtagcc ggccagcctc
     7141 gcagagcagg attcccgttg agcaccgcca ggtgcgaata agggacagtg aagaaggaac
     7201 acccgctcgc gggtgggcct acttcaccta tcctgcccgg ctgacgccgt tggatacacc
     7261 aaggaaagtc tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat
     7321 ataccgaaaa aatcgctata atgaccccga agcagggtta tgcagcggaa aagatccgtc
     7381 gacctgcagg catgctaaac cgatacaatt aaaggctcct tttggagcct ttttttttgg
     7441 aggcgaactc ccttatgcgg gtggcgcaga atgccaggac ccttgtcatt ccacgtcaat
     7501 tcatgcgcct tttcacctcg tactgtcctg ccaaacacaa gcaacggagg tacg
//
