LOCUS       62056_13550        9668 bp DNA     circular SYN 15-MAR-2021
DEFINITION  Knockout vector pKO7393, complete sequence.
ACCESSION   MW659890
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9668)
  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 9668)
  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..9668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..26)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(34..50)
                     /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(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"
     primer_bind     6499..6515
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             7763..8554
                     /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 cagcagtccg tcatcaacca ggccctggcc
     6601 ggcgcacggc tcacccccgc acaggtcgac gcggtcgagg cacacggcac cggcacccgg
     6661 ctcggcgacc ccatcgaggc ccaggccctg atcggcacct acggcgcgga acgcgcagag
     6721 gaccgcccgc tctggctcgg ctcgctgaag tcgaacatcg gccactccca ggcggcggcc
     6781 ggggccggcg gcatcatcaa gatggtcatg gcgatgcggc acggcgtgct tcccaagacc
     6841 ctgcacgtcg acgaacccag cagccacgtc gactggtcgg cgggcaccgt ggcgctcctc
     6901 accgaggaac gcccgtggcc ccggaccgac gaaccgcgcc gcgccggcgt gtcctccttc
     6961 ggcgtcagcg gcaccaacgc ccacgtcatc ctggaagagg cgcccgcgcc cgacccggcc
     7021 accccggaca gcgacgctcc ggccaccgac cggccgacgc ccgccatcac cccctgggtg
     7081 ctgtccgcca agacccccga cgcgctgcgc gcacaggccg accggctgcg gtccgccgtg
     7141 gcggcctcca ccgacttcga ccccacgggc gtcggcttct cgctggccac cacccgctcc
     7201 ctcttcgagc accgcgccgt gatcctcgcc gaggaccgcg ccaccaccat cgaccggctc
     7261 accgccctcg ccgaaggaca cggcgccccc ggtgtcatgt ccggcgaggc cgcaccgcac
     7321 cggctggcct tcctgttctc ggggcagggc tcgcagcgtc ttggcatggg ccgtgaactg
     7381 gccgcgcgct atgaggtgtt cgcggaggcc ctggacgccg tactcgacga gtgcgacccg
     7441 cgcgtgcgtg aggtcctgtt cggcgaggac gccgacgcgc tgaacgagac gggggtgacc
     7501 cagcccgcgc tgttcgcggt cgaggtggcc ctcttccgcc tgcttgagtc gtggggtgtg
     7561 cgcccggacg tgctggccgg tcactcgatt cggtacccgg ggatccgatt ggacagcaag
     7621 cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa
     7681 ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagct ctgatcaaga
     7741 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     7801 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     7861 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     7921 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     7981 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     8041 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     8101 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     8161 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     8221 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     8281 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     8341 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     8401 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     8461 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     8521 catcgccttc tatcgccttc ttgacgagtt cttctgaatt attaacgctt acaatttcct
     8581 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatct ctagagtcga
     8641 cctgcaaggt atcccggctc aaggtcagcc acgcgttcca ctcgccgctg atggagccga
     8701 tgctcgacga cttccgccgg gtacttgagg gcgtctcgca cgaggcgccg tcgctcccga
     8761 tcgtgtcgac gctgaccggt gtccgcgcga gcgccgatga gctgacgtcg ccggactact
     8821 gggtgcggca cgtgcgcgag gccgtgcgct tccacggcgc ggtggcggag ctgcgcgagc
     8881 agggcgtcga cgtcttcctg gaggtcggcc ccggtggcgt gctctccggg ctcgggcagg
     8941 tcggcgcccc cgaggccgcc ttcgtgcccg ccctccgtgg cgaccgcccc gaactggccg
     9001 cgctgaccac cgccatcggg cgactccacg tacgcggcgt cccgctggac ctcgccgccc
     9061 tcttcgacgg cagcggggcc cagcgggtcg acctgccgac gtacgcgttc cagcgcgagc
     9121 cctactggct cgacgccccg gcggccgagg agagcaccgg gcgtacctca ccggaggagg
     9181 cgcggttctg ggaggccgtg gaggagggtg accccgccga cctcgcccgt acgctgggcg
     9241 tgtcgtccga cgacccgctg agcgcggtgc tgccccgcct gtccgcctgg cgcaggaagc
     9301 agcgcgaccg gctgacggcc gacggctggc ggtaccgggt cgactggcgt gcggtggacg
     9361 ccgggacgcg ccggccggcc ggcgcctggc tggtcgtgac gccggccggt gacgagtgcg
     9421 ccgccgagtg ggtaagggcc tcgctgacgc gtaacggcgc ggacgaggcg catgtcctgc
     9481 ccgtggacct ggcctcgggg gcggggacgc tcgactgggc ggagcggctg gcgggcctgc
     9541 cgcggctgag cggtgtggtc tcgctgctcg gtctcgccga cgcgggggag gggaccgtac
     9601 ccgcgggcgt cgcggccacg atcggtctgc tgcgggcgct cgacgacatg gacgcaccgc
     9661 tgtgaatt
//
