LOCUS       40924_19886        3851 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Chloramphenicol resistance FRT vector pFCM1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3851)
  AUTHORS   Choi KH, Gaynor JB, White KG, Lopez C, Bosio CM, Karkhoff-Schweizer 
            RR, Schweizer HP.
  TITLE     A Tn7-based broad-range bacterial cloning and expression system
  JOURNAL   Nat. Methods 2 (6), 443-448 (2005)
  PUBMED    15908923
REFERENCE   2  (bases 1 to 3851)
  AUTHORS   Choi KH, Gaynor J, Karkhoff-Schweizer RR, Schweizer HP.
  TITLE     Improved FRT cassettes and Flp expression vectors
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 3851)
  AUTHORS   Choi KH, Gaynor J, Karkhoff-Schweizer RR, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-APR-2004) Microbiology, Immunology and Pathology, 
            Colorado State University, Fort Collins, CO 80523, USA
REFERENCE   4  (bases 1 to 3851)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 3851)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Methods"; date: "2005"; volume: "2"; issue: "6"; pages: "443-448"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (12-APR-2004) Microbiology, Immunology and Pathology, Colorado State
            University, Fort Collins, CO 80523, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(399..455)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     protein_bind    464..511
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        596..698
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             699..1355
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     misc_feature    1497..1544
                     /note="FRT; Flp recombinase target site"
     protein_bind    1497..1544
                     /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    1561..1617
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1630..1646)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1654..1670)
                     /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(1678..1708)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1723..1744)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2032..2620)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2794..3651)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3652..3756)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttgcatgc ctgcaggtcg
      421 actctagagg atccccgggt accgagctcg aattggggat cttgaagtac ctattccgaa
      481 gttcctattc tctagaaagt ataggaactt cagagcgctt ttgaagctga tgtgcttaaa
      541 aacttactca atggaataat tctagatggg ccaacttttg gcgaaaatga gacgttgatc
      601 ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt
      661 ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
      721 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
      781 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
      841 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
      901 atgctcatcc ggagttccgt atggcaatga aagacggtga gctggtgata tgggatagtg
      961 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
     1021 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
     1081 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca
     1141 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
     1201 cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
     1261 cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat
     1321 tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt
     1381 gcccttaaac gcctggtgct acgcctgaat aagtgataat aagcggatga atggcagaaa
     1441 ttcgaaagca aattcgaccc atccattgct gttgacaaag ggaatcaggg gatcttgaag
     1501 ttcctattcc gaagttccta ttctctagaa agtataggaa cttcagagcg cttttgaagc
     1561 taattcgagc tcggtacccg gggatcctct agagtcgacc tgcaggcatg caagcttggc
     1621 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1681 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1741 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     1801 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     1861 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1921 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1981 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2041 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2101 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2161 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2221 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2281 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2341 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2401 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2461 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2521 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2581 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2641 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2701 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2761 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2821 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2881 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2941 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3001 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3061 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3121 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3181 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3241 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3301 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3361 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3421 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3481 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3541 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3601 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3661 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3721 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3781 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     3841 gccctttcgt c
//
