LOCUS       40924_19691        3905 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Apramycin resistance FRT vector pFAP1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3905)
  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 3905)
  AUTHORS   Choi K-H., Gaynor JB, White KG, Lopez C, Bosio CM, 
            Karkhoff-Schweizer RR, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-JUL-2005) Microbiology, Immunology and Pathology, 
            Colorado State University, Fort Collins, CO 80523, USA
REFERENCE   3  (bases 1 to 3905)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3905)
  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: "Submitted 
            (12-JUL-2005) Microbiology, Immunology and Pathology, Colorado State
            University, Fort Collins, CO 80523, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3905
                     /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)."
     CDS             complement(613..1413)
                     /codon_start=1
                     /label=ApmR
                     /note="aminoglycoside 3-N-acetyltransferase type IV"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     misc_feature    1551..1598
                     /label=FRT, Flp recombinase target site
                     /note="FRT, Flp recombinase target site"
     protein_bind    1551..1598
                     /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    1615..1671
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1684..1700)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1708..1724)
                     /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(1732..1762)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1777..1798)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2086..2674)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2848..3705)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3706..3810)
                     /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 tctagataat tcggcttctc caacgtcatc tcgttctccg
      601 ctcatgagct cagccaatcg actggcgagc ggcatcgcat tcttcgcatc ccgcctctgg
      661 cggatgcagg aagatcaacg gatctcggcc cagttgaccc agggctgtcg ccacaatgtc
      721 gcgggagcgg atcaaccgag caaaggcatg accgactgga ccttccttct gaaggctctt
      781 ctccttgagc cacctgtccg ccaaggcaaa gcgctcacag cagtggtcat tctcgagata
      841 atcgacgcgt accaacttgc catcctgaag aatggtgcag tgtctcggca ccccataggg
      901 aacctttgcc atcaactcgg caagatgcag cgtcgtgttg gcatcgtgtc ccacgccgag
      961 gagaagtacc tgcccatcga gttcatggac acgggcgacc gggcttgcag gcgagtgagg
     1021 tggcaggggc aatggatcag agatgatctg ctctgcctgt ggccccgctg ccgcaaaggc
     1081 aaatggatgg gcgctgcgct ttacatttgg caggcgccag aatgtgtcag agacaactcc
     1141 aaggtccggt gtaacgggcg acgtggcagg atcgaacggc tcgtcgtcca gacctgacca
     1201 cgagggcatg acgagcgtcc ctcccggacc cagcgcagca cgcagggcct cgatcagtcc
     1261 aagtggccca tcttcgaggg gccggacgct acggaaggag ctgtggacca gcagcacacc
     1321 gccgggggta accccaaggt tgagaagctg accgatgagc tcggcttttc gccattcgta
     1381 ttgcacgaca ttgcactcca ccgctgatga catcagtcga tcatagcacg atcaacggca
     1441 ctgttgcaaa tagtcggtgg tgataaactt atcatcccct tttgctgatg gagctgcaca
     1501 tgaaaagccg aattatccat tgctgttgac aaagggaatc aggggatctt gaagttccta
     1561 ttccgaagtt cctattctct agaaagtata ggaacttcag agcgcttttg aagctaattc
     1621 gagctcggta cccggggatc ctctagagtc gacctgcagg catgcaagct tggcgtaatc
     1681 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     1741 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     1801 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     1861 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     1921 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     1981 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2041 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2101 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2161 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2221 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2281 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2341 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2401 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2461 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2521 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2581 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2641 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2701 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2761 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2821 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2881 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     2941 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3001 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3061 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3121 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3181 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3241 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3301 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3361 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3421 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3481 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3541 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     3601 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     3661 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     3721 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     3781 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     3841 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     3901 tcgtc
//
