LOCUS       62056_13325        5834 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5834)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    21..39
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     CDS             478..1269
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    complement(1463..1496)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     CDS             complement(1524..2231)
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     protein_bind    complement(2278..2294)
                     /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(2302..2331)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     misc_feature    complement(2344..2362)
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     rep_origin      complement(2375..2758)
                     /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(2831..3688)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3689..3793)
                     /label=AmpR promoter
     CDS             complement(4415..4783)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(4816..4925)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     terminator      5602..5688
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      5780..5807
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
ORIGIN
        1 agattgcagc attacacgtc ctgtctctta tacacatctt tgagcgattg tgtaggctgg
       61 agctgcttcg aagttcctat actttctaga gaataggaac ttcggaatag gaacttcaag
      121 atcccctcac gctgccgcaa gcactcaggg cgcaagggct gctaaaggaa gcggaacacg
      181 tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta ctgggctatc
      241 tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg
      301 cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc agctggggcg
      361 ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaagg
      421 atctgatggc gcaggggatc aagatctgat caagagacag gatgaggatc gtttcgcatg
      481 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
      541 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
      601 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
      661 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
      721 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
      781 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
      841 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
      901 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
      961 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     1021 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     1081 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     1141 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     1201 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     1261 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     1321 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     1381 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     1441 accccagctt caaaagcgct ctgaagttcc tatactttct agagaatagg aacttcggaa
     1501 taggaactaa ggaggatatt ttacttgtac agctcgtcca tgccgccggt ggagtggcgg
     1561 ccctcggcgc gttcgtactg ttccacgatg gtgtagtcct cgttgtggga ggtgatgtcc
     1621 aacttgatgt tgacgttgta ggcgccgggc agctgcacgg gcttcttggc cttgtaggtg
     1681 gtcttgacct cagcgtcgta gtggccgccg tccttcagct tcagcctctg cttgatctcg
     1741 cccttcaggg cgccgtcctc ggggtacatc cgctcggagg aggcctccca gcccatggtc
     1801 ttcttctgca ttacggggcc gtcggagggg aagttggtgc cgcgcagctt caccttgtag
     1861 atgaactcgc cgtcctgcag ggaggagtcc tgggtcacgg tcaccacgcc gccgtcctcg
     1921 aagttcatca cgcgctccca cttgaagccc tcggggaagg acagcttcaa gtagtcgggg
     1981 atgtcggcgg ggtgcttcac gtaggccttg gagccgtaca tgaactgagg ggacaggatg
     2041 tcccaggcga agggcagggg gccacccttg gtcaccttca gcttggcggt ctgggtgccc
     2101 tcgtaggggc ggccctcgcc ctcgccctcg atctcgaact cgtggccgtt cacggagccc
     2161 tccatgtgca ccttgaagcg catgaactcc ttgatgatgg ccatgttatc ctcctcgccc
     2221 ttgctcacca tgaatactgt ttcctgtgtg aaatatccgc tcatgagaca ttccacattg
     2281 ttatccgctc acaattccac acattatacg agccggatga ttaattgtca acatatggac
     2341 catagatgtg tataagagac agggctaatt cccatgtcag ccgttaagtg ttcctgtgtc
     2401 actgaaaatt gctttgagag gctctaaggg cttctcagtg cgttacatcc ctggcttgtt
     2461 gtccacaacc gttaaacctt aaaagcttta aaagccttat atattctttt ttttcttata
     2521 aaacttaaaa ccttagaggc tatttaagtt gctgatttat attaatttta ttgttcaaac
     2581 atgagagctt agtacgtgaa acatgagagc ttagtacgtt agccatgaga gcttagtacg
     2641 ttagccatga gggtttagtt cgttaaacat gagagcttag tacgttaaac atgagagctt
     2701 agtacgtgaa acatgagagc ttagtacgta ctatcaacag gttgaactgc ggatcttgcg
     2761 gccgcaaaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2821 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2881 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     2941 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3001 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3061 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3121 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3181 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3241 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3301 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3361 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3421 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3481 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3541 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3601 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3661 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     3721 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3781 taggggttcc gcgcacattt ccccgaaaag tgccacctgc atcgagccga ccaggctttc
     3841 cacgcccgcg tgccgctcca tgtcgttcgc gcggttctcg gaaacgcgct gccgcgtttc
     3901 gtgattgtca cgctcaagcc cgtagtcccg ttcgagcgtc gcgcagaggt cagcgagggc
     3961 gcggtaggcc cgatacggct catggatggt gtttcgggtc gggtgaatct tgttgatggc
     4021 gatatggatg tgcaggttgt cggtgtcgtg atgcacggca ctgacgcgct gatgctcggc
     4081 gaagccaagc ccagcgcaga tgcggtcctc aatcgcgcgc aacgtctccg cgtcgggctt
     4141 ctctcccgcg cggaagctaa ccagcaggtg ataggtcttg tcggcctcgg aacgggtgtt
     4201 gccgtgctgg gtcgccatca cctcggccat gacagcgggc agggtgtttg cctcgcagtt
     4261 cgtgacgcgc acgtgaccca ggcgctcggt cttgccttgc tcgtcggtga tgtacttcac
     4321 cagctccgcg aagtcgctct tcttgatgga gcgcatgggg acgtgcttgg caatcacgcg
     4381 caccccccgg ccgttttagc ggctaaaaaa gtcatggctc tgccctcggg cggaccacgc
     4441 ccatcatgac cttgccaagc tcgtcctgct tctcttcgat cttcgccagc agggcgagga
     4501 tcgtggcatc accgaaccgc gccgtgcgcg ggtcgtcggt gagccagagt ttcagcaggc
     4561 cgcccaggcg gcccaggtcg ccattgatgc gggccagctc gcggacgtgc tcatagtcca
     4621 cgacgcccgt gattttgtag ccctggccga cggccagcag gtaggccgac aggctcatgc
     4681 cggccgccgc cgccttttcc tcaatcgctc ttcgttcgtc tggaaggcag tacaccttga
     4741 taggtgggct gcccttcctg gttggcttgg tttcatcagc catccgcttg ccctcatctg
     4801 ttacgccggc ggtagccggc cagcctcgca gagcaggatt cccgttgagc accgccaggt
     4861 gcgaataagg gacagtgaag aaggaacacc cgctcgcggg tgggcctact tcacctatcc
     4921 tgcccggctg acgccgttgg atacaccaag gaaagtctac acgaaccctt tggcaaaatc
     4981 ctgtatatcg tgcgaaaaag gatggatata ccgaaaaaat cgctataatg accccgaagc
     5041 agggttatgc agcggaaaag cgctgcttcc ctgctgtttt gtggaatatc taccgactgg
     5101 aaacaggcaa atgcaggaaa ttactgaact gaggggacag gcgagagacg atgccaaaga
     5161 gctacaccga cgagctggcc gagtgggttg aatcccgcgc ggccaagaag cgccggcgtg
     5221 atgaggctgc ggttgcgttc ctggcggtga gggcggatgt cgaggcggcg ttagcgtccg
     5281 gctatgcgct cgtcaccatt tgggagcaca tgcgggaaac ggggaaggtc aagttctcct
     5341 acgagacgtt ccgctcgcac gccaggcggc acatcaaggc caagcccgcc gatgtgcccg
     5401 caccgcaggc caaggctgcg gaacccgcgc cggcacccaa gacgccggag ccacggcggc
     5461 cgaagcaggg gggcaaggct gaaaagccgg cccccgctgc ggccccgacc ggcttcacct
     5521 tcaacccaac accggacaaa aaatggcccc ccgatggtag tgtggggtct ccccatgcga
     5581 gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     5641 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg
     5701 gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact
     5761 gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc gtggccagtg
     5821 ccaagcttgc atgc
//
