LOCUS       62056_13335        5809 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 5809)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5809
                     /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"
     promoter        complement(2278..2306)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     misc_feature    complement(2319..2337)
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     rep_origin      complement(2350..2733)
                     /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(2806..3663)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3664..3768)
                     /label=AmpR promoter
     CDS             complement(4390..4758)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(4791..4900)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     terminator      5577..5663
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      5755..5782
                     /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 ttccacacat
     2281 tatacgagcc gatgattaat tgtcaacata tggaccatag atgtgtataa gagacagggc
     2341 taattcccat gtcagccgtt aagtgttcct gtgtcactga aaattgcttt gagaggctct
     2401 aagggcttct cagtgcgtta catccctggc ttgttgtcca caaccgttaa accttaaaag
     2461 ctttaaaagc cttatatatt cttttttttc ttataaaact taaaacctta gaggctattt
     2521 aagttgctga tttatattaa ttttattgtt caaacatgag agcttagtac gtgaaacatg
     2581 agagcttagt acgttagcca tgagagctta gtacgttagc catgagggtt tagttcgtta
     2641 aacatgagag cttagtacgt taaacatgag agcttagtac gtgaaacatg agagcttagt
     2701 acgtactatc aacaggttga actgcggatc ttgcggccgc aaaaattaaa aatgaagttt
     2761 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2821 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2881 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2941 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3001 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3061 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3121 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3181 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3241 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3301 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3361 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3421 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3481 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3541 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3601 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3661 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3721 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3781 aaaagtgcca cctgcatcga gccgaccagg ctttccacgc ccgcgtgccg ctccatgtcg
     3841 ttcgcgcggt tctcggaaac gcgctgccgc gtttcgtgat tgtcacgctc aagcccgtag
     3901 tcccgttcga gcgtcgcgca gaggtcagcg agggcgcggt aggcccgata cggctcatgg
     3961 atggtgtttc gggtcgggtg aatcttgttg atggcgatat ggatgtgcag gttgtcggtg
     4021 tcgtgatgca cggcactgac gcgctgatgc tcggcgaagc caagcccagc gcagatgcgg
     4081 tcctcaatcg cgcgcaacgt ctccgcgtcg ggcttctctc ccgcgcggaa gctaaccagc
     4141 aggtgatagg tcttgtcggc ctcggaacgg gtgttgccgt gctgggtcgc catcacctcg
     4201 gccatgacag cgggcagggt gtttgcctcg cagttcgtga cgcgcacgtg acccaggcgc
     4261 tcggtcttgc cttgctcgtc ggtgatgtac ttcaccagct ccgcgaagtc gctcttcttg
     4321 atggagcgca tggggacgtg cttggcaatc acgcgcaccc cccggccgtt ttagcggcta
     4381 aaaaagtcat ggctctgccc tcgggcggac cacgcccatc atgaccttgc caagctcgtc
     4441 ctgcttctct tcgatcttcg ccagcagggc gaggatcgtg gcatcaccga accgcgccgt
     4501 gcgcgggtcg tcggtgagcc agagtttcag caggccgccc aggcggccca ggtcgccatt
     4561 gatgcgggcc agctcgcgga cgtgctcata gtccacgacg cccgtgattt tgtagccctg
     4621 gccgacggcc agcaggtagg ccgacaggct catgccggcc gccgccgcct tttcctcaat
     4681 cgctcttcgt tcgtctggaa ggcagtacac cttgataggt gggctgccct tcctggttgg
     4741 cttggtttca tcagccatcc gcttgccctc atctgttacg ccggcggtag ccggccagcc
     4801 tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa taagggacag tgaagaagga
     4861 acacccgctc gcgggtgggc ctacttcacc tatcctgccc ggctgacgcc gttggataca
     4921 ccaaggaaag tctacacgaa ccctttggca aaatcctgta tatcgtgcga aaaaggatgg
     4981 atataccgaa aaaatcgcta taatgacccc gaagcagggt tatgcagcgg aaaagcgctg
     5041 cttccctgct gttttgtgga atatctaccg actggaaaca ggcaaatgca ggaaattact
     5101 gaactgaggg gacaggcgag agacgatgcc aaagagctac accgacgagc tggccgagtg
     5161 ggttgaatcc cgcgcggcca agaagcgccg gcgtgatgag gctgcggttg cgttcctggc
     5221 ggtgagggcg gatgtcgagg cggcgttagc gtccggctat gcgctcgtca ccatttggga
     5281 gcacatgcgg gaaacgggga aggtcaagtt ctcctacgag acgttccgct cgcacgccag
     5341 gcggcacatc aaggccaagc ccgccgatgt gcccgcaccg caggccaagg ctgcggaacc
     5401 cgcgccggca cccaagacgc cggagccacg gcggccgaag caggggggca aggctgaaaa
     5461 gccggccccc gctgcggccc cgaccggctt caccttcaac ccaacaccgg acaaaaaatg
     5521 gccccccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa
     5581 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     5641 acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc
     5701 ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg
     5761 ccatcctgac ggatggcctt tttgcgtggc cagtgccaag cttgcatgc
//
