LOCUS       62056_13555        9983 bp DNA     circular SYN 15-MAR-2021
DEFINITION  Knockout vector pKO7395, complete sequence.
ACCESSION   MW659891
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9983)
  AUTHORS   Han X, Li Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-FEB-2021) College of plant protection, Hebei 
            Agricultural University, Lekai south street No. 2596, Baoding, Hebei
            071000, China
REFERENCE   2  (bases 1 to 9983)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (24-FEB-2021) College of plant protection, Hebei Agricultural 
            University, Lekai south street No. 2596, Baoding, Hebei 071000, 
            China"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9983
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(58..88)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(103..124)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(412..1000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             1204..2004
                     /codon_start=1
                     /label=ApmR
                     /note="aminoglycoside 3-N-acetyltransferase type IV"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     CDS             complement(2947..4386)
                     /codon_start=1
                     /label=pSG5 Rep
                     /note="replication initiator protein from the Streptomyces 
                     ghanaensis plasmid pSG5"
                     /translation="MWKPGEATWGNTCRCNNVHTCPWCMSRILAVRGSNVQLAADGLAD
                     AGYGLHLGTNTLRHFERMAFGTVRKGMRHGLVAVLHDGWKGAYGSSGRRWRTMRDDFGI
                     IGYERAFEDTFGWGSGWHLHWHTLWVTREVLGPDAQAAFRDALAGAWAAGVESAGGYTV
                     SETCDRPGCSCEGKGHGTDVRPLNGADAADGDAGKQARYLYKDGDKTKGGVAKIGLELA
                     GQNFKAGRGDDRMGPLDLGDAAAAELQRLRRPGPFVEKYREREFGVFQVRKHYRSQNLN
                     RLIKELGIQQDVRTEEEITDDTEGLVAIAVIPAYIWYRYIARVAGRRLDLIKVAETYGL
                     PGVRRLVESWGLVWGKDVLDPPAPEAPAAPGDLDADQMRFEVMSEEEAAFREARRKANE
                     ARTEELAASLDRVRQPKKEAIRPTISLRKRLKPKPVTVDVKTPPPGAASPVCRRCKGKL
                     APVLQPWGQHPGDCLRVDTAVA"
     oriT            5553..5662
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             8082..8873
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
ORIGIN
        1 cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
       61 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
      121 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
      181 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
      241 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
      301 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
      361 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
      421 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
      481 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
      541 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
      601 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
      661 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
      721 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
      781 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
      841 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
      901 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
      961 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1021 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1081 atcaaaaagg atcttcacct agatcctttt ggttcatgtg cagctccatc agcaaaaggg
     1141 gatgataagt ttatcaccac cgactatttg caacagtgcc gttgatcgtg ctatgatcga
     1201 ctgatgtcat cagcggtgga gtgcaatgtc gtgcaatacg aatggcgaaa agccgagctc
     1261 atcggtcagc ttctcaacct tggggttacc cccggcggtg tgctgctggt ccacagctcc
     1321 ttccgtagcg tccggcccct cgaagatggg ccacttggac tgatcgaggc cctgcgtgct
     1381 gcgctgggtc cgggagggac gctcgtcatg ccctcgtggt caggtctgga cgacgagccg
     1441 ttcgatcctg ccacgtcgcc cgttacaccg gaccttggag ttgtctctga cacattctgg
     1501 cgcctgccaa atgtaaagcg cagcgcccat ccatttgcct ttgcggcagc ggggccacag
     1561 gcagagcaga tcatctctga tccattgccc ctgccacctc actcgcctgc aagcccggtc
     1621 gcccgtgtcc atgaactcga tgggcaggta cttctcctcg gcgtgggaca cgatgccaac
     1681 acgacgctgc atcttgccga gttgatggca aaggttccct atggggtgcc gagacactgc
     1741 accattcttc aggatggcaa gttggtacgc gtcgattatc tcgagaatga ccactgctgt
     1801 gagcgctttg ccttggcgga caggtggctc aaggagaaga gccttcagaa ggaaggtcca
     1861 gtcggtcatg cctttgctcg gttgatccgc tcccgcgaca ttgtggcgac agccctgggt
     1921 caactgggcc gagatccgtt gatcttcctg catccgccag aggcgggatg cgaagaatgc
     1981 gatgccgctc gccagtcgat tggctgagct catgagcgga gaacgagatg acgttggagg
     2041 ggcaaggtcg cgctgattgc tggggcaaca cgtggagcgg atcggggatt gtctttcttc
     2101 agctcgctga tgatatgctg acgctcaatg ccgtttggcc tccgactaac gaaaatcccg
     2161 catttggacg gctgatccga ttggcacggc ggacggcgaa tggcggagca gacgctcgtc
     2221 cgggggcaat gagatatgaa aaagcctgaa ctcaccgcga cgtatcgatg tcgaggttcc
     2281 tcaggggagc caccccagag aagccctcgg agctgagcgg agctatttcc aaagccatgc
     2341 cagctagaga cagtgcacac cgccaagcat ctgcaaaacc ctcgcctgga gggaaagtgc
     2401 aatgtacgta cgctggtttc cctccagaaa gggattctgc ggcttctcaa ctttggaaga
     2461 agaggcggaa cgaactgctg tttcagccta ctcatgtgag gaggctggtc ctttaccctg
     2521 atgcccgagg gcatcagggt aaaggaccag cctcgccaac taggagcgct accaaggcga
     2581 acacaaccca ggacttggta gaacccttcg ccggagaagt tcagactcat cggcatgagc
     2641 acccctcgcc gcgcactcgg caccggtccg tcgaccacca ccaggacgtc gttgtcgacg
     2701 tcggccccgc ggctcctgcc cgccgaacgc gtcgtcgtcg acggcctggt gctcatcgac
     2761 gagcacccgg agccaggtga aaagcgccgg cggacgctcg gactgggcgc gggattccag
     2821 cagtaaccca ggtccgccgg ccacctcacg gcaggcagac cctcggcttt cgcgccggga
     2881 ccgccatgag accgccaccc ggatgtccgg ggtggcggtc tcatgggcgg tccctcagcg
     2941 gccctacgcg accgctgtgt cgacgcggag gcagtctccg gggtgctggc cccagggctg
     3001 gagtaccggg gcgagcttgc ccttgcagcg gcggcacacg ggcgacgcgg cgccgggcgg
     3061 gggagtcttg acgtcgacgg tgaccggctt cggcttcagg cgcttccgaa ggctgatcgt
     3121 cgggcggatc gcctccttct tcggctgacg cactcggtcg agcgaagcgg ccagctcctc
     3181 cgtgcgggcc tcgttggcct tccggcgcgc ctcgcggaaa gcagcttcct cctcggacat
     3241 gacctcgaac ctcatctggt cagcgtcaag gtcgcccggc gcggccggcg cttccggggc
     3301 gggcgggtcc aggacgtcct tgccccacac caggccccag gactcgacga gccgccggac
     3361 gcccggtagg ccgtacgtct cggcgacctt gatgaggtcg aggcgacgtc cggcgacgcg
     3421 ggcgatgtat cggtaccaga tgtaggccgg gatgaccgcg atggcgacca ggccctcggt
     3481 gtcgtcggtg atctcctcct cggtgcggac gtcctgctgg atgccgagtt ccttgatcag
     3541 ccggttcagg ttctgcgacc ggtagtgctt gcggacctgg aagacgccga actcgcgctc
     3601 gcggtacttc tcgacgaacg ggccgggccg acgaagccgc tgcagctcgg cggccgccgc
     3661 gtcgcccagg tcgagcggtc ccatgcggtc gtcgccgcga ccggccttga agttctgtcc
     3721 ggccagctcc aggccgatct tggcgacgcc gcccttggtc ttgtcgccgt ccttgtagag
     3781 gtagcgggcc tgcttgcccg catcgccgtc agcggcgtcc gcgccgttga gtgggcgcac
     3841 gtcggtgccg tggcccttgc cctcacagga gcaaccgggc cggtcgcacg tctcgctgac
     3901 ggtgtagccg cccgcggatt cgaccccggc ggcccaggct ccggcgagtg cgtcgcggaa
     3961 cgcggcctgg gcgtccgggc cgagcacctc gcgggtgacc cagagcgtgt gccagtgcag
     4021 gtgccagccg gagccccagc cgaaggtgtc ctcgaaggcc cgctcgtagc cgatgatccc
     4081 gaagtcgtcg cgcatcgtgc gccagcggcg gccggacgag ccgtacgcgc ccttccagcc
     4141 gtcgtgcaag accgcgacca ggccgtgccg cattcccttg cggacggtgc cgaacgccat
     4201 gcgctcgaag tggcgcaacg tgttcgtgcc aaggtgcagc ccgtacccgg cgtccgcgag
     4261 accgtcggcg gcgagctgca cgttcgagcc ccgtacggcc aggatgcggc tcatgcacca
     4321 cgggcaggtg tggacgttgt tgcagcggca cgtgttgccc cacgtcgcct cgcccggctt
     4381 ccacatcagc tcggccgtcc cggcagtgag ccgggtcccg cagcccttga acgcctcgtt
     4441 cagcgacacc gtctggtgcc ggtcccgccg ggcgaaccgc tcgtcgcgcg ggtcctcccg
     4501 cccggctgtc gcggcaccct cgtttggggt agaacccgtt ccagttacag cgctctgacc
     4561 tgcagtggac ggagattttc ccttactact aaagcccgcg tccggattac ccgctgtagt
     4621 cgtgcttgct acgctgcgtg actggtccgc aatgagacgc tttgcgcgct ttcggcaggc
     4681 gtccgagcag tagattttgg ggcgcttccc ggggatgtgg acgatcgggg tgccgcagtg
     4741 gcacttcggt ccggcggggc gcggtggtgt cgacgcgctg ttctctcgta cgctcgtcac
     4801 agagcaaacg tcctcactcg gcatgctgcg ccggttcggg ggcggcgagc ccgggaggcc
     4861 aatcccgggc tcgtgccatt tctgggtcct gttgatcctg gcattggtgt ggccgttcat
     4921 tgcccctgct cgctcctgac gcgccgatag acgtccgata cgcccggtgc tggtgggatt
     4981 tgataggtcg gaagaagccc cgcccgggcc tgggcggggc ttcctgtgcg tcaggacctc
     5041 ctcgtcgtga gcctcttcgg cctatggacg gagtgacctc gtgatccgtt acagccgcgc
     5101 gcgctcgcgt agagcggtct catcagttcc acgaacggtc ctcttcgcag atcagggcgt
     5161 tggggcggag tctcaccaag gactacgtct gctggcgatt tccgttacac cccgggcggt
     5221 ggccggcgca cacgcgcgcc cgcgttgggc agtgcagaaa gtgcagaaac ctaggcgctg
     5281 atggtccagg tccacggttc gtcgtcggcg gcggcgcggg cggcggcgtc ggccagggcg
     5341 cgggcgagac cggctacggc gggcttgatg cgccggttgc gggcgacctt gagcagctag
     5401 tatgcaggtc gacggatctt ttccgctgca taaccctgct tcggggtcat tatagcgatt
     5461 ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag ggttcgtgta
     5521 gactttcctt ggtgtatcca acggcgtcag ccgggcagga taggtgaagt aggcccaccc
     5581 gcgagcgggt gttccttctt cactgtccct tattcgcacc tggcggtgct caacgggaat
     5641 cctgctctgc gaggctggcc ggcatgagcc tgtcggccta cctgctggcc gtcggccagg
     5701 gctacaaaat cacgggcgtc gtggactatg agcacgtccg cgagctggcc cgcatcaatg
     5761 gcgacctggg ccgcctgggc ggcctgctga aactctggct caccgacgac ccgcgcacgg
     5821 cgcggttcgg tgatgccacg atcctcgccc tgctggcgaa gatcgaagag aagcaggacg
     5881 agcttggcaa ggtcatgatg ggcgtggtcc gcccgagggc agagccatga cttttttagc
     5941 cgctaaaacg gccggggggt gcgcgtgatt gccaagcacg tccccatgcg ctccatcaag
     6001 aagagcgact tcgcggagct ggtgaagtac atcaccgacg agcaaggcaa gaccgatccc
     6061 cggggacctg cataagatct acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg
     6121 tctccgacct gatgcagctc tcgcagggcg aagaatctcg tgctttcagc ttcgatgtag
     6181 gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt
     6241 atgttgatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg
     6301 aatttatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     6361 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     6421 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     6481 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttgggctg caggtcgact
     6541 ctagaggatc cgcggccgcg cgcgatatcc acctcgtgtc gcaccctgac ctcgacctgc
     6601 tcgacgtggc cgccacactc gcggacaccc gggccgccct cgaccaccgg gcggtcgtcg
     6661 tcggccagga ccgcgacggg atgctctcgg ccctcaccgc gctcgccgag gggcgggccg
     6721 cgaacggtct cgtctccgga atcgcggccg ggcgccggac ggcgttcctg ttctccggcc
     6781 agggcgcgca gcggctcggt atgggccgcg acctgtacga ccggttcccg gtgttcgccg
     6841 acgcgctcga cgaggtctgc gcccacctgg acacccacct ggcactccac ccggagaccc
     6901 gccccgaggg tgaccggccg ctgcgggacc tgctcttcgg cgacgacgcc gaggcgctga
     6961 accagaccgc cgtgacccag cccgcgctgt tcgcggtcga ggtggcgctg ttccgcctcc
     7021 tggagtcctg gggcgtgcgc cccgagacgc tggcgggcca ctcgatcggc gagctggccg
     7081 cggcccacgt ggccggcgtc tggtcactcg ccgacgccgc cgcggtggtg gcgagccggg
     7141 gccgcctcat gcaggccctg cccgagggcg gcgcgatgtt cgccgtccag gcgaccgagg
     7201 ccgagatcct gccggagctg accgacaccg tcggcatcgc cgcggtgaac ggcccgaact
     7261 cggtggtcgt ctccggtgtc gccgccgacg ccgaggccat cgccgagcgt tggcgggcga
     7321 gcggacggaa ggtgacacgg ctgcgcgtca gccacgcctt ccactcgccg ctcatggaac
     7381 cgatgctcga cgacttccgc cgcgtcctcg aagaggtctc ctacgagacg ccgaagatcc
     7441 ccgtggtgtc caccctgacc ggcgtccgcg ccaccgccga ggagctcacg tccccggagt
     7501 actgggtgcg ccacgtacgg gaggcggtgc ggttcgccga cgcggtcggc accctgcgcg
     7561 aacagggcat ggacgccttc ctggagatcg ggccccgcgg agtgctcacc gcgctgggcc
     7621 aggagagcgc gcccgacgcc gcgttcgtcc ccgccctgcg cggcgaccgg cacgaggtgc
     7681 tcgaactggc gacggccgcg gccaggctgc acgtgctcgg gccggcgtcg actggaccgc
     7741 gctgaccgcc ggcggcagcc acatcgacct gcccacctac gccttccagc ggaagcggtt
     7801 ctggcccacc ctggccgagg cccccgcggg cgacgtctcc ggagccgtcg acgcgcggtt
     7861 ctgggcggcc gtggccgagg gcgacctcgg tgagctggcg gatgaacttc ggtacccggg
     7921 gatccgattg gacagcaagc gaaccggaat tgccagctgg ggcgccctct ggtaaggttg
     7981 ggaagccctg caaagtaaac tggatggctt tcttgccgcc aaggatctga tggcgcaggg
     8041 gatcaagctc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     8101 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     8161 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     8221 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     8281 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     8341 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     8401 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     8461 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     8521 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     8581 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     8641 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     8701 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     8761 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     8821 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgaatta
     8881 ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     8941 accgcatctc tagagtcgac ctgcaaccgc gtcacctggc aaccggtgag cgagcccacc
     9001 gcggcgctca cgggtacctg gctcgtggtc agcagggacg gcgccgacga ggaatccgtc
     9061 acggccgcgc tgggcaggca cggggcgcga gtggtccccg tacggaccgg tgaagcggtg
     9121 gaccgcgaag ccctcgcggc ccggatgacg accggcgagc ccgtcgcggg cgtactgtcg
     9181 ctgttcgcac tcgacgccga actgccctcc ggcggcctgt acgcgaccct cgccctcgtc
     9241 caggcgctca gcgacatcgg cctcgccgcc ccgctgtggt gcgcgacacg cggcggagtc
     9301 gccgtcggcg gctccgacag ccccgccgac cccggccagg cgatggtgtg gggactcggc
     9361 agggtcgtcg cactcgaaca ccccgaacgc tggggaggcc tcgtcgacct gcccgccacc
     9421 ctggacgaac gcgccggcgc ccgcctcgcg ggtgtcctgg caggaacgca gggcgaggac
     9481 caggtcgcac tccgcgccgg cggggtgtac ggccgccggc tcgcgccggt gacgtcggca
     9541 cgcctgggcg actgggaccc caccggcacc gtcctcgtca ccggcggcac cggctcgctc
     9601 ggcggccacg tcgccaagtg gctggtggcc aagggagccg agcacctgct cctcaccagc
     9661 aggcgcggcc tcgaggccga cggcgcgaag gaactgcacg ccgaactgac cgggctggga
     9721 gcccacgtca cgatcgcggc ctgcgacgtc gccgaccgcg acgccctcgc ggacctgctg
     9781 gcgtccgtcc cggaggagtt cccgctcacc gcggtcgtgc acgccgccgg caccctggac
     9841 gacggcgtgc tgtccgcact caccccggag cggctcgacg ccgtgctgcg gccgaaggcc
     9901 gtcgccgcgg ccaacctcga cgagctgacc cgggacctgg acctctcggc gttcgtcatg
     9961 ttctcctccg tgtccggcga att
//
