LOCUS       62056_13330        5801 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 5801)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5801
                     /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..2240)
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     promoter        complement(2270..2298)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     misc_feature    complement(2311..2329)
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     rep_origin      complement(2342..2725)
                     /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(2798..3655)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3656..3760)
                     /label=AmpR promoter
     CDS             complement(4382..4750)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(4783..4892)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     terminator      5569..5655
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      5747..5774
                     /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 tgccgagagt gatcccggcg
     1561 gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagg
     1621 gcggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg
     1681 gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc
     1741 ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac gttgtggctg
     1801 ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc
     1861 ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg
     1921 tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg
     1981 gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg
     2041 taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg
     2101 aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac
     2161 ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc
     2221 tcctcgccct tgctcaccat gaatactgtt tcctgtgtga aattccacac attatacgag
     2281 ccgatgatta attgtcaaca tatggaccat agatgtgtat aagagacagg gctaattccc
     2341 atgtcagccg ttaagtgttc ctgtgtcact gaaaattgct ttgagaggct ctaagggctt
     2401 ctcagtgcgt tacatccctg gcttgttgtc cacaaccgtt aaaccttaaa agctttaaaa
     2461 gccttatata ttcttttttt tcttataaaa cttaaaacct tagaggctat ttaagttgct
     2521 gatttatatt aattttattg ttcaaacatg agagcttagt acgtgaaaca tgagagctta
     2581 gtacgttagc catgagagct tagtacgtta gccatgaggg tttagttcgt taaacatgag
     2641 agcttagtac gttaaacatg agagcttagt acgtgaaaca tgagagctta gtacgtacta
     2701 tcaacaggtt gaactgcgga tcttgcggcc gcaaaaatta aaaatgaagt tttaaatcaa
     2761 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2821 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     2881 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     2941 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3001 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3061 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3121 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3181 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3241 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3301 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3361 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3421 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3481 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3541 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3601 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     3661 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3721 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     3781 cacctgcatc gagccgacca ggctttccac gcccgcgtgc cgctccatgt cgttcgcgcg
     3841 gttctcggaa acgcgctgcc gcgtttcgtg attgtcacgc tcaagcccgt agtcccgttc
     3901 gagcgtcgcg cagaggtcag cgagggcgcg gtaggcccga tacggctcat ggatggtgtt
     3961 tcgggtcggg tgaatcttgt tgatggcgat atggatgtgc aggttgtcgg tgtcgtgatg
     4021 cacggcactg acgcgctgat gctcggcgaa gccaagccca gcgcagatgc ggtcctcaat
     4081 cgcgcgcaac gtctccgcgt cgggcttctc tcccgcgcgg aagctaacca gcaggtgata
     4141 ggtcttgtcg gcctcggaac gggtgttgcc gtgctgggtc gccatcacct cggccatgac
     4201 agcgggcagg gtgtttgcct cgcagttcgt gacgcgcacg tgacccaggc gctcggtctt
     4261 gccttgctcg tcggtgatgt acttcaccag ctccgcgaag tcgctcttct tgatggagcg
     4321 catggggacg tgcttggcaa tcacgcgcac cccccggccg ttttagcggc taaaaaagtc
     4381 atggctctgc cctcgggcgg accacgccca tcatgacctt gccaagctcg tcctgcttct
     4441 cttcgatctt cgccagcagg gcgaggatcg tggcatcacc gaaccgcgcc gtgcgcgggt
     4501 cgtcggtgag ccagagtttc agcaggccgc ccaggcggcc caggtcgcca ttgatgcggg
     4561 ccagctcgcg gacgtgctca tagtccacga cgcccgtgat tttgtagccc tggccgacgg
     4621 ccagcaggta ggccgacagg ctcatgccgg ccgccgccgc cttttcctca atcgctcttc
     4681 gttcgtctgg aaggcagtac accttgatag gtgggctgcc cttcctggtt ggcttggttt
     4741 catcagccat ccgcttgccc tcatctgtta cgccggcggt agccggccag cctcgcagag
     4801 caggattccc gttgagcacc gccaggtgcg aataagggac agtgaagaag gaacacccgc
     4861 tcgcgggtgg gcctacttca cctatcctgc ccggctgacg ccgttggata caccaaggaa
     4921 agtctacacg aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat ggatataccg
     4981 aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc ggaaaagcgc tgcttccctg
     5041 ctgttttgtg gaatatctac cgactggaaa caggcaaatg caggaaatta ctgaactgag
     5101 gggacaggcg agagacgatg ccaaagagct acaccgacga gctggccgag tgggttgaat
     5161 cccgcgcggc caagaagcgc cggcgtgatg aggctgcggt tgcgttcctg gcggtgaggg
     5221 cggatgtcga ggcggcgtta gcgtccggct atgcgctcgt caccatttgg gagcacatgc
     5281 gggaaacggg gaaggtcaag ttctcctacg agacgttccg ctcgcacgcc aggcggcaca
     5341 tcaaggccaa gcccgccgat gtgcccgcac cgcaggccaa ggctgcggaa cccgcgccgg
     5401 cacccaagac gccggagcca cggcggccga agcagggggg caaggctgaa aagccggccc
     5461 ccgctgcggc cccgaccggc ttcaccttca acccaacacc ggacaaaaaa tggccccccg
     5521 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga
     5581 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     5641 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
     5701 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
     5761 acggatggcc tttttgcgtg gccagtgcca agcttgcatg c
//
