LOCUS       62056_6495        6205 bp DNA     circular SYN 03-NOV-2020
DEFINITION  Suicide vector pCKD100, complete sequence.
ACCESSION   MW039600
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6205)
  AUTHORS   Nazareno ES, Acharya B, Dumenyo CK.
  TITLE     A mini-Tn5-derived transposon with reportable and selectable markers
            enables rapid generation and screening of insertional mutants in 
            Gram-negative bacteria
  JOURNAL   Lett. Appl. Microbiol. (2020) In press
REFERENCE   2  (bases 1 to 6205)
  AUTHORS   Nazareno ES, Acharya B, Dumenyo CK.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 6205)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Lett. Appl.
            Microbiol. (2020) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-SEP-2020) Agricultural "
FEATURES             Location/Qualifiers
     source          1..6205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   7..25
                     /label=Tn5 inverted repeat
                     /note="Tn5 inverted repeat"
     misc_feature    26..59
                     /label=rare-cutting endonuclease multiple cloning site I
                     /note="rare-cutting endonuclease multiple cloning site I"
     rep_origin      complement(84..472)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(610..1266)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     misc_feature    1341..1360
                     /label=rare-cutting endonuclease multiple cloning site II
                     /note="rare-cutting endonuclease multiple cloning site II"
     CDS             complement(1537..2328)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             complement(2396..3109)
                     /label=GFPuv
                     /note="GFP variant optimized for excitation by UV light"
     gene            complement(3114..3165)
                     /gene="atpE"
                     /label=atpE
     regulatory      complement(3114..3165)
                     /gene="atpE"
                     /regulatory_class="ribosome_binding_site"
     repeat_region   3181..3198
                     /rpt_family="Tn5"
     oriT            3915..4024
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(4686..6113)
                     /label=Tn5 transposase
                     /note="transposase from the bacterial Tn5 transposon
                     (Reznikoff, 1993)"
ORIGIN
        1 tctagactgt ctcttgatca gatctggcca cctaggccgg ccgcgatcgc cggcgcgcca
       61 tgcatgtcga cggtaccccc ccatgtcagc cgttaagtgt tcctgtgtca ctcaaaattg
      121 ctttgagagg ctctaagggc ttctcagtgc gttacatccc tggcttgttg tccacaaccg
      181 ttaaacctta aaagctttaa aagccttata tattcttttt tttcttataa aacttaaaac
      241 cttagaggct atttaagttg ctgatttata ttaattttat tgttcaaaca tgagagctta
      301 gtacgtgaaa catgagagct tagtacgtta gccatgagag cttagtacgt tagccatgag
      361 ggtttagttc gttaaacatg agagcttagt acgttaaaca tgagagctta gtacgtgaaa
      421 catgagagct tagtacgtac tatcaacagg ttgaactgct gatcttcaga tcctctacgc
      481 cggacgcatc gtggccgggg ttcgaaatcg atgagctcgc gaatttctgc cattcatccg
      541 cttattatca cttattcagg cgtagcaacc aggcgtttaa gggcaccaat aactgcctta
      601 aaaaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct
      661 gccgacatgg aagccatcac agacggcatg atgaacctga atcgccagcg gcatcagcac
      721 cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat
      781 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa
      841 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
      901 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
      961 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
     1021 cagctcaccg tctttcattg ccatacggaa ttccggatga gcattcatca ggcgggcaag
     1081 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
     1141 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
     1201 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
     1261 ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     1321 tgatcttatt tcaagagctc ttaattaatt taaatctaga ggggtgggcg aagaactcca
     1381 gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg attccgaagc
     1441 ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg
     1501 cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata
     1561 gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc
     1621 ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg
     1681 gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat
     1741 gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg
     1801 cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc
     1861 atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc
     1921 ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc
     1981 catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac
     2041 ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca
     2101 aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca gttcattcag
     2161 ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa
     2221 cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc
     2281 cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc
     2341 tcatcctgtc tcttgatcag atccactagt ccagtgtggt ggaattgaat tcttatttgt
     2401 agagctcatc catgccatgt gtaatcccag cagcagttac aaactcaaga aggaccatgt
     2461 ggtcacgctt ttcgttggga tctttcgaaa gggcagattg tgtcgacagg taatggttgt
     2521 ctggtaaaag gacagggcca tcgccaattg gagtattttg ttgataatgg tctgctagtt
     2581 gaacggatcc atcttcaatg ttgtggcgaa ttttgaagtt agctttgatt ccattctttt
     2641 gtttgtctgc cgtgatgtat acattgtgtg agttatagtt gtactcgagt ttgtgtccga
     2701 gaatgtttcc atcttcttta aaatcaatac cttttaactc gatacgatta acaagggtat
     2761 caccttcaaa cttgacttca gcacgcgtct tgtagttccc gtcatctttg aaagatatag
     2821 tgcgttcctg tacataacct tcgggcatgg cactcttgaa aaagtcatgc cgtttcatat
     2881 gatctgggta tcttgaaaag cattgaacac cataagagaa agtagtgaca agtgttggcc
     2941 atggaacagg tagttttcca gtagtgcaaa taaatttaag ggtaagtttt ccgtatgttg
     3001 catcaccttc accctctcca ctgacagaaa atttgtgccc attaacatca ccatctaatt
     3061 caacaagaat tgggacaact ccagtgaaaa gttcttctcc tttactcatt atgagtctcc
     3121 agtttgtttc agttaaaacg tagtagtgtt ggtaaattac tcgagtctag agcggccgct
     3181 acttgtgtat aagagtcaga attctactgt ttgggtgtat gagctatttg caaaggaaat
     3241 tggtgatgac aaagctcggc gctatttgaa gaaaatcgac tatggcaacg ccgatccttt
     3301 ttgtccggtg ttgggttgaa ggtgaagccg gtcggggccg cagcgggggc cggcttttca
     3361 gccttgcccc cctgcttcgg ccgccgtggc tccggcgtct tgggtgccgg cgcgggttcc
     3421 gcagccttgg cctgcggtgc gggcacatcg gcgggcttgg ccttgatgtg ccgcctggcg
     3481 tgcgagcgga acgtctcgta ggagaacttg accttccccg tttcccgcat gtgctcccaa
     3541 atggtgacga gcgcatagcc ggacgctaac gccgcctcga catccgccct caccgccagg
     3601 aacgcaaccg cagcctcatc acgccggcgc ttcttggccg cgcgggattc aacccactcg
     3661 gccagctcgt cggtgtagct ctttggcatc gtctctcgcc tgtcccctca gttcagtaat
     3721 ttcctgcatt tgcctgtttc cagtcggtag atattccaca aaacagcagg gaagcagcgc
     3781 ttttccgctg cataaccctg cttcggggtc attatagcga ttttttcggt atatccatcc
     3841 tttttcgcac gatatacagg attttgccaa agggttcgtg tagactttcc ttggtgtatc
     3901 caacggcgtc agccgggcag gataggtgaa gtaggcccac ccgcgagcgg gtgttccttc
     3961 ttcactgtcc cttattcgca cctggcggtg ctcaacggga atcctgctct gcgaggctgg
     4021 ccggctaccg ccggcgtaac agatgagggc aagcggatgg ctgatgaaac caagccaacc
     4081 aggaagggca gcccacctat caaggtgtac tgccttccag acgaacgaag agcgattgag
     4141 gaaaaggcgg cggcggccgg catgagcctg tcggcctacc tgctggccgt cggccagggc
     4201 tacaaaatca cgggcgtcgt ggactatgag cacgtccgcg agctggcccg catcaatggc
     4261 gacctgggcc gcctgggcgg cctgctgaaa ctctggctca ccgacgaccc gcgcacggcg
     4321 cggttcggtg atgccacgat ccgggcaacg ttgttgccat tgctgcaggc gcagaactgg
     4381 taggtatgga agatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt
     4441 gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc
     4501 gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg
     4561 ttgggcgcca tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac
     4621 ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgtcg agcgggggcc
     4681 gatcagatct tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta
     4741 ctttgcaggg cttcccaacc ttcccagagg gcgccccagc tggcaattcc ggttcgcttg
     4801 ctgtccataa aaccgcccag tctagctatc gccatgtaag cccactgcaa gctacctgct
     4861 ttctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg
     4921 gtcagcaccg tttctgcgga ctggctttct acgtgttccg cttcctttag cagcccttgc
     4981 gccctgagtg cttgcggcag cgtgaagctt tctctgagct gtaacagcct gaccgcaaca
     5041 aacgagagga tcgagaccat ccgctccaga ttatccggct cctccatgcg ttgcctctcg
     5101 gctcctgctc cggttttcca tgccttatgg aactcctcga tccgccagcg atgggtataa
     5161 atgtcgatga cgcgcaaggc ttgggctagc gactcgaccg gttcgctggt cagcaacaac
     5221 catttcaacg gggtctcacc cttgggcggg ttaatctcct cggccagcac cgcgttgagc
     5281 gtgatattcc cctgttttag cgtgatgcgc ccactgcgca ggctcaagct cgccttgcgg
     5341 gctggtcgat ttttacgttt accgcgttta tccaccacgc ccttttgcgg aatgctgatc
     5401 tgatagccac ccaactccgg ttggttcttc agatggtcgt acagatacaa cccagactct
     5461 acgtccttgc gtgggtgctt ggagcgcacc acgaagcgct cgttatgcgc cagtttgtcc
     5521 tgcagataag catgaatatc ggcttcgcgg tcacagaccg caatcacgtt gctcatcatg
     5581 ctgcccatgc gtaaccggct agttgcggcg gctgccagcc atttgccact ctccttttca
     5641 tccgcatcgg cagggtcatc cgggcgcatc caccactcct gatgcagtaa tcctacggtg
     5701 cggaatgtgg tggcctcgag caagagaacg gagtgaaccc accatccgcg ggatttatcc
     5761 tgaatagagc ccagcttgcc aagctcttcg gcgacctggt ggcgataact caaagaggtg
     5821 gtgtcctcaa tggccagcag ttcgggaaac tcctgagcca acttgactgt ttgcatggcg
     5881 ccagcctttc tgatcgcctc ggcagaaacg ttgggattgc ggataaatcg gtaagcgcct
     5941 tcctgcatgg cttcactacc ctctgatgag atggttattg atttaccaga atattttgcc
     6001 aattgggcgg cgacgttaac caagcgggca gtacggcgag gatcacccag cgccgccgaa
     6061 gagaacacag atttagccca gtcggccgca cgatgaagag cagaagttat catgaacgtt
     6121 accatgttag gcaggtcaca tggaagatca gatcctggaa aacgggaaag gttccgttca
     6181 ggacgctact tgtgtacggg atcgg
//
