LOCUS       40924_11041        3043 bp DNA     circular SYN 17-DEC-2018
DEFINITION  PCR amplification vector pCLF2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3043)
  AUTHORS   Santiviago CA, Blondel CJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-MAR-2010) Laboratorio de Microbiologia, Departamento 
            de Bioquimica y Biologia Molecular, Facultad de Ciencias Quimicas y 
            Farmaceuticas, Universidad de Chile, Avenida Vicuna Mackenna 20, 
            Santiago, Region Metropolitana, Chile
REFERENCE   2  (bases 1 to 3043)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 3043)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (31-MAR-2010) Laboratorio de Microbiologia, Departamento de 
            Bioquimica y Biologia Molecular, Facultad de Ciencias Quimicas y 
            Farmaceuticas, Universidad de Chile, Avenida Vicuna Mackenna 20, 
            Santiago, Region Metropolitana, Chile"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     31..50
                     /label=priming site P1
                     /note="priming site P1"
     primer_bind     32..51
                     /label=priming site P3
                     /note="priming site P3"
     protein_bind    complement(51..98)
                     /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)."
     misc_feature    51..96
                     /note="FLP recognition target site; FRT"
     promoter        273..375
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             376..1014
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyl transferase"
                     /label=cat
                     /note="confers chloramphenicol resistance"
                     /protein_id="ADZ54044.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRCLMNTTVLR"
     gene            376..1014
                     /gene="cat"
                     /label=cat
     primer_bind     complement(470..492)
                     /label=priming site c3
                     /note="priming site c3"
     primer_bind     903..922
                     /label=priming site c1
                     /note="priming site c1"
     protein_bind    complement(1150..1183)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     promoter        1203..1221
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1227..1246)
                     /label=priming site P2
                     /note="priming site P2"
     rep_origin      complement(1265..1653)
                     /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(1758..2615)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(2616..2720)
                     /label=AmpR promoter
     terminator      2811..2897
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2989..3016
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
ORIGIN
        1 agattgcagc attacacgtc ttgagcgatt gtgtaggctg gagctgcttc gaagttccta
       61 tactttctag agaataggaa cttcggaata ggaacttcaa gatcccctca cgctgccgca
      121 agcactcagg gcgcaagggc tgctaaagga agcggaacac gtagaaagcc agtccggatc
      181 ctgacggaag atcacttcgc agaataaata aatcctggtg tccctgttga taccgggaag
      241 ccctgggcca acttttggcg aaaatgagac gttgatcggc acgtaagagg ttccaacttt
      301 caccataatg aaataagatc actaccgggc gtattttttg agttgtcgag attttcagga
      361 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa
      421 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
      481 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
      541 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attacgtatg
      601 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc
      661 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag
      721 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct
      781 aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt
      841 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa
      901 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt
      961 tgtgatggct tccatgtcgg cagatgctta atgaatacaa cagtactgcg atgagtggca
     1021 gggcggggcg taaggcgcgc catttaaatg aagctgcaga atcgttttcc gggacgccgg
     1081 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccagcttcaa
     1141 aagcgctctg aagttcctat actttctaga gaataggaac ttcggaatag gaattccgaa
     1201 attaatacga ctcactatag ggagacctaa ggaggatatt catatggacc atggctaatt
     1261 cccatgtcag ccgttaagtg ttcctgtgtc actcaaaatt gctttgagag gctctaaggg
     1321 cttctcagtg cgttacatcc ctggcttgtt gtccacaacc gttaaacctt aaaagcttta
     1381 aaagccttat atattctttt ttttcttata aaacttaaaa ccttagaggc tatttaagtt
     1441 gctgatttat attaatttta ttgttcaaac atgagagctt agtacgtgaa acatgagagc
     1501 ttagtacgtt agccatgaga gcttagtacg ttagccatga gggtttagtt cgttaaacat
     1561 gagagcttag tacgttaaac atgagagctt agtacgtgaa acatgagagc ttagtacgta
     1621 ctatcaacag gttgaactgc tgatcttcag atcctctacg ccggacgcat cgtggccgga
     1681 tcttgcggcc gcaaaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     1741 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     1801 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     1861 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     1921 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     1981 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     2041 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     2101 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     2161 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     2221 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     2281 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     2341 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     2401 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     2461 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     2521 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     2581 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     2641 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     2701 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgcatc gatggccccc
     2761 cgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac
     2821 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     2881 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag
     2941 ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc
     3001 tgacggatgg cctttttgcg tggccagtgc caagcttgca tgc
//
