LOCUS       pKD3                2842 bp ds-DNA     circular SYN 28-APR-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pKD3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2842)
  AUTHORS   Datsenko KA, Wanner BL
  TITLE     One-step inactivation of chromosomal genes in Escherichia coli K-12 
            using PCR products.
  JOURNAL   Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6640-5.
  PUBMED    10829079
REFERENCE   2  (bases 1 to 2842)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 28, 2020 from SnapGene Server 1.1.58
            //gt12365.cn
FEATURES             Location/Qualifiers
     source          1..2842
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      107..193
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      285..312
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     protein_bind    complement(390..437)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(454..1113)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1028..1047
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1114..1216)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(1322..1355)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     rep_origin      1404..1792
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(1894..2754)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2517..2536
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2755..17)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgcatcgat ggcccccgat
       61 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaatgaaa
      121 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
      181 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg
      241 gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac
      301 ggatggcctt tttgcgtggc cagtgccaag cttgcatgca gattgcagca ttacacgtct
      361 tgagcgattg tgtaggctgg agctgcttcg aagttcctat actttctaga gaataggaac
      421 ttcggaatag gaacttcatt taaatggcgc gccttacgcc ccgccctgcc actcatcgca
      481 gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg
      541 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt
      601 gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact
      661 cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc
      721 caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc
      781 gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta
      841 acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca tacgtaattc
      901 cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt
      961 atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt
     1021 acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc
     1081 aacggtggta tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct
     1141 cgacaactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
     1201 tcttacgtgc cgatcaacgt ctcattttcg ccaaaagttg gcccagggct tcccggtatc
     1261 aacagggaca ccaggattta tttattctgc gaagtgatct tccgtcacag gtaggcgcgc
     1321 cgaagttcct atactttcta gagaatagga acttcggaat aggaactaag gaggatattc
     1381 atatggacca tggctaattc ccatgtcagc cgttaagtgt tcctgtgtca ctcaaaattg
     1441 ctttgagagg ctctaagggc ttctcagtgc gttacatccc tggcttgttg tccacaaccg
     1501 ttaaacctta aaagctttaa aagccttata tattcttttt tttcttataa aacttaaaac
     1561 cttagaggct atttaagttg ctgatttata ttaattttat tgttcaaaca tgagagctta
     1621 gtacgtgaaa catgagagct tagtacgtta gccatgagag cttagtacgt tagccatgag
     1681 ggtttagttc gttaaacatg agagcttagt acgttaaaca tgagagctta gtacgtgaaa
     1741 catgagagct tagtacgtac tatcaacagg ttgaactgct gatcttcaga tcctctacgc
     1801 cggacgcatc gtggccggat cttgcggccg caaaaattaa aaatgaagtt ttaaatcaat
     1861 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     1921 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     1981 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     2041 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     2101 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     2161 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     2221 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     2281 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     2341 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     2401 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     2461 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     2521 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     2581 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     2641 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     2701 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     2761 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     2821 tgaatgtatt tagaaaaata aa
//