LOCUS       62056_13545        8779 bp DNA     circular SYN 15-MAR-2021
DEFINITION  Knockout vector pKO7392, complete sequence.
ACCESSION   MW659889
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8779)
  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 8779)
  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..8779
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            2271..2380
                     /label=oriT
                     /note="incP origin of transfer"
     primer_bind     3217..3233
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4033..4824
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4912..4923)
                     /codon_start=1
                     /label=WELQut site
                     /note="WELQut protease recognition and cleavage site"
                     /translation="WELQ"
     primer_bind     complement(5507..5523)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5531..5547)
                     /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(5555..5585)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5600..5621)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5909..6497)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             6701..7501
                     /codon_start=1
                     /label=ApmR
                     /note="aminoglycoside 3-N-acetyltransferase type IV"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     CDS             complement(join(8444..8779,1..1104))
                     /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"
ORIGIN
        1 ggccggcgct tccggggcgg gcgggtccag gacgtccttg ccccacacca ggccccagga
       61 ctcgacgagc cgccggacgc ccggtaggcc gtacgtctcg gcgaccttga tgaggtcgag
      121 gcgacgtccg gcgacgcggg cgatgtatcg gtaccagatg taggccggga tgaccgcgat
      181 ggcgaccagg ccctcggtgt cgtcggtgat ctcctcctcg gtgcggacgt cctgctggat
      241 gccgagttcc ttgatcagcc ggttcaggtt ctgcgaccgg tagtgcttgc ggacctggaa
      301 gacgccgaac tcgcgctcgc ggtacttctc gacgaacggg ccgggccgac gaagccgctg
      361 cagctcggcg gccgccgcgt cgcccaggtc gagcggtccc atgcggtcgt cgccgcgacc
      421 ggccttgaag ttctgtccgg ccagctccag gccgatcttg gcgacgccgc ccttggtctt
      481 gtcgccgtcc ttgtagaggt agcgggcctg cttgcccgca tcgccgtcag cggcgtccgc
      541 gccgttgagt gggcgcacgt cggtgccgtg gcccttgccc tcacaggagc aaccgggccg
      601 gtcgcacgtc tcgctgacgg tgtagccgcc cgcggattcg accccggcgg cccaggctcc
      661 ggcgagtgcg tcgcggaacg cggcctgggc gtccgggccg agcacctcgc gggtgaccca
      721 gagcgtgtgc cagtgcaggt gccagccgga gccccagccg aaggtgtcct cgaaggcccg
      781 ctcgtagccg atgatcccga agtcgtcgcg catcgtgcgc cagcggcggc cggacgagcc
      841 gtacgcgccc ttccagccgt cgtgcaagac cgcgaccagg ccgtgccgca ttcccttgcg
      901 gacggtgccg aacgccatgc gctcgaagtg gcgcaacgtg ttcgtgccaa ggtgcagccc
      961 gtacccggcg tccgcgagac cgtcggcggc gagctgcacg ttcgagcccc gtacggccag
     1021 gatgcggctc atgcaccacg ggcaggtgtg gacgttgttg cagcggcacg tgttgcccca
     1081 cgtcgcctcg cccggcttcc acatcagctc ggccgtcccg gcagtgagcc gggtcccgca
     1141 gcccttgaac gcctcgttca gcgacaccgt ctggtgccgg tcccgccggg cgaaccgctc
     1201 gtcgcgcggg tcctcccgcc cggctgtcgc ggcaccctcg tttggggtag aacccgttcc
     1261 agttacagcg ctctgacctg cagtggacgg agattttccc ttactactaa agcccgcgtc
     1321 cggattaccc gctgtagtcg tgcttgctac gctgcgtgac tggtccgcaa tgagacgctt
     1381 tgcgcgcttt cggcaggcgt ccgagcagta gattttgggg cgcttcccgg ggatgtggac
     1441 gatcggggtg ccgcagtggc acttcggtcc ggcggggcgc ggtggtgtcg acgcgctgtt
     1501 ctctcgtacg ctcgtcacag agcaaacgtc ctcactcggc atgctgcgcc ggttcggggg
     1561 cggcgagccc gggaggccaa tcccgggctc gtgccatttc tgggtcctgt tgatcctggc
     1621 attggtgtgg ccgttcattg cccctgctcg ctcctgacgc gccgatagac gtccgatacg
     1681 cccggtgctg gtgggatttg ataggtcgga agaagccccg cccgggcctg ggcggggctt
     1741 cctgtgcgtc aggacctcct cgtcgtgagc ctcttcggcc tatggacgga gtgacctcgt
     1801 gatccgttac agccgcgcgc gctcgcgtag agcggtctca tcagttccac gaacggtcct
     1861 cttcgcagat cagggcgttg gggcggagtc tcaccaagga ctacgtctgc tggcgatttc
     1921 cgttacaccc cgggcggtgg ccggcgcaca cgcgcgcccg cgttgggcag tgcagaaagt
     1981 gcagaaacct aggcgctgat ggtccaggtc cacggttcgt cgtcggcggc ggcgcgggcg
     2041 gcggcgtcgg ccagggcgcg ggcgagaccg gctacggcgg gcttgatgcg ccggttgcgg
     2101 gcgaccttga gcagctagta tgcaggtcga cggatctttt ccgctgcata accctgcttc
     2161 ggggtcatta tagcgatttt ttcggtatat ccatcctttt tcgcacgata tacaggattt
     2221 tgccaaaggg ttcgtgtaga ctttccttgg tgtatccaac ggcgtcagcc gggcaggata
     2281 ggtgaagtag gcccacccgc gagcgggtgt tccttcttca ctgtccctta ttcgcacctg
     2341 gcggtgctca acgggaatcc tgctctgcga ggctggccgg catgagcctg tcggcctacc
     2401 tgctggccgt cggccagggc tacaaaatca cgggcgtcgt ggactatgag cacgtccgcg
     2461 agctggcccg catcaatggc gacctgggcc gcctgggcgg cctgctgaaa ctctggctca
     2521 ccgacgaccc gcgcacggcg cggttcggtg atgccacgat cctcgccctg ctggcgaaga
     2581 tcgaagagaa gcaggacgag cttggcaagg tcatgatggg cgtggtccgc ccgagggcag
     2641 agccatgact tttttagccg ctaaaacggc cggggggtgc gcgtgattgc caagcacgtc
     2701 cccatgcgct ccatcaagaa gagcgacttc gcggagctgg tgaagtacat caccgacgag
     2761 caaggcaaga ccgatccccg gggacctgca taagatctac gtctgtcgag aagtttctga
     2821 tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc gcagggcgaa gaatctcgtg
     2881 ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg
     2941 gtttctacaa agatcgttat gttgatcggc actttgcatc ggccgcgctc ccgattccgg
     3001 aagtgcttga cattggggaa tttatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     3061 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     3121 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     3181 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc
     3241 ttgggctgca ggtcgactct agaggatccg cggccgcgcg cgatatcgct cggcacagac
     3301 ggagaacccc ggccgcttca cgctggtgga cctcgacgac accgaggaac ccaccgacgc
     3361 cgtgcgcgcc gcactggcct ccggcgaacc ccagaccacc gtccgggcag gccaggtgcg
     3421 cgtgccccgg ctcgcccgca ccaccgacga cgacgcgctc accccgcccg ccgacggacc
     3481 gtggcgcctg gacagccgac agcgcggctc cctcgacggt ctcgccctgg tgccgcaccc
     3541 cgacgccgcc gcgcccctcg cgcccggcga ggtgcgtatc gccgtacgcg ccgcgggcct
     3601 caacttccgc gacgtcctga tcgccctcgg ctcctacccc ggcgaggccg acatgggcag
     3661 cgaggtcgcc ggcgtggtca ccgaggtcgg caccggggtc accgatctgg cggtgggcga
     3721 ccgcgtcctc ggcatggtca ccaagggctt cggcccgctc gccgtcaccg accgccgact
     3781 ggtcgccccg atgcccgccg gatggtcctt cgcccgcgcc gcgtccgtgc cactggtctt
     3841 cctcaccgcc tactacgcct cggtacccgg ggatccgatt ggacagcaag cgaaccggaa
     3901 ttgccagctg gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct
     3961 ttcttgccgc caaggatctg atggcgcagg ggatcaagct ctgatcaaga gacaggatga
     4021 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4081 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4141 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4201 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4261 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4321 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4381 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4441 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4501 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4561 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4621 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4681 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4741 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4801 tatcgccttc ttgacgagtt cttctgaatt attaacgctt acaatttcct gatgcggtat
     4861 tttctcctta cgcatctgtg cggtatttca caccgcatct ctagagtcga cctgcaactc
     4921 ccacctggcg tcctcgcgcg acaccgcctt cgcggcacgg ttcaccgccg agaccgacgg
     4981 ccgcggcgtg gacgtggtgc tgaactccct ggtacgggaa ttcgtcgacg cctccctcga
     5041 actgctgccc gacggcggtc ggttcatcga gctcggcaag gccgacgtac gcgacaccgg
     5101 caccctgccc tccggcgtcg agtacgccac gttcgacctg acacagccgg gccccgaccg
     5161 cttccaggag atgctcgtcg aggtgctcgc cctgttcgag gcgggcgccc tcacccccct
     5221 gccggtgcgc gcctgggacg tccgccgcgc acccgaggcg ttccgccacc tcagccaggc
     5281 ccgccacgtc ggcaaggtcg tgctcacggt gcccgcgccc ctcgacccgg cccgccccgt
     5341 cctggtgacc ggcggcctcg gcaccctcgg cgccctggtg gcccggcacc tcgtcgccga
     5401 acacggcgtg cgcaacctcg tactcgcggg ccgcaaggga ccggcgaccg agggcgcggc
     5461 ggcactgcgc gacgaactgg agacactggg cggaattcgt aatcatgtca tagctgtttc
     5521 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5581 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5641 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5701 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5761 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5821 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5881 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5941 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6001 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6061 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6121 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6181 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6241 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6301 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6361 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6421 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6481 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6541 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6601 tccttttggt tcatgtgcag ctccatcagc aaaaggggat gataagttta tcaccaccga
     6661 ctatttgcaa cagtgccgtt gatcgtgcta tgatcgactg atgtcatcag cggtggagtg
     6721 caatgtcgtg caatacgaat ggcgaaaagc cgagctcatc ggtcagcttc tcaaccttgg
     6781 ggttaccccc ggcggtgtgc tgctggtcca cagctccttc cgtagcgtcc ggcccctcga
     6841 agatgggcca cttggactga tcgaggccct gcgtgctgcg ctgggtccgg gagggacgct
     6901 cgtcatgccc tcgtggtcag gtctggacga cgagccgttc gatcctgcca cgtcgcccgt
     6961 tacaccggac cttggagttg tctctgacac attctggcgc ctgccaaatg taaagcgcag
     7021 cgcccatcca tttgcctttg cggcagcggg gccacaggca gagcagatca tctctgatcc
     7081 attgcccctg ccacctcact cgcctgcaag cccggtcgcc cgtgtccatg aactcgatgg
     7141 gcaggtactt ctcctcggcg tgggacacga tgccaacacg acgctgcatc ttgccgagtt
     7201 gatggcaaag gttccctatg gggtgccgag acactgcacc attcttcagg atggcaagtt
     7261 ggtacgcgtc gattatctcg agaatgacca ctgctgtgag cgctttgcct tggcggacag
     7321 gtggctcaag gagaagagcc ttcagaagga aggtccagtc ggtcatgcct ttgctcggtt
     7381 gatccgctcc cgcgacattg tggcgacagc cctgggtcaa ctgggccgag atccgttgat
     7441 cttcctgcat ccgccagagg cgggatgcga agaatgcgat gccgctcgcc agtcgattgg
     7501 ctgagctcat gagcggagaa cgagatgacg ttggaggggc aaggtcgcgc tgattgctgg
     7561 ggcaacacgt ggagcggatc ggggattgtc tttcttcagc tcgctgatga tatgctgacg
     7621 ctcaatgccg tttggcctcc gactaacgaa aatcccgcat ttggacggct gatccgattg
     7681 gcacggcgga cggcgaatgg cggagcagac gctcgtccgg gggcaatgag atatgaaaaa
     7741 gcctgaactc accgcgacgt atcgatgtcg aggttcctca ggggagccac cccagagaag
     7801 ccctcggagc tgagcggagc tatttccaaa gccatgccag ctagagacag tgcacaccgc
     7861 caagcatctg caaaaccctc gcctggaggg aaagtgcaat gtacgtacgc tggtttccct
     7921 ccagaaaggg attctgcggc ttctcaactt tggaagaaga ggcggaacga actgctgttt
     7981 cagcctactc atgtgaggag gctggtcctt taccctgatg cccgagggca tcagggtaaa
     8041 ggaccagcct cgccaactag gagcgctacc aaggcgaaca caacccagga cttggtagaa
     8101 cccttcgccg gagaagttca gactcatcgg catgagcacc cctcgccgcg cactcggcac
     8161 cggtccgtcg accaccacca ggacgtcgtt gtcgacgtcg gccccgcggc tcctgcccgc
     8221 cgaacgcgtc gtcgtcgacg gcctggtgct catcgacgag cacccggagc caggtgaaaa
     8281 gcgccggcgg acgctcggac tgggcgcggg attccagcag taacccaggt ccgccggcca
     8341 cctcacggca ggcagaccct cggctttcgc gccgggaccg ccatgagacc gccacccgga
     8401 tgtccggggt ggcggtctca tgggcggtcc ctcagcggcc ctacgcgacc gctgtgtcga
     8461 cgcggaggca gtctccgggg tgctggcccc agggctggag taccggggcg agcttgccct
     8521 tgcagcggcg gcacacgggc gacgcggcgc cgggcggggg agtcttgacg tcgacggtga
     8581 ccggcttcgg cttcaggcgc ttccgaaggc tgatcgtcgg gcggatcgcc tccttcttcg
     8641 gctgacgcac tcggtcgagc gaagcggcca gctcctccgt gcgggcctcg ttggccttcc
     8701 ggcgcgcctc gcggaaagca gcttcctcct cggacatgac ctcgaacctc atctggtcag
     8761 cgtcaaggtc gcccggcgc
//
