LOCUS       40924_27379        5695 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pL0E_0-R, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5695)
  AUTHORS   Hochrein L, Machens F, Gremmels J, Schulz K, Messerschmidt K, 
            Mueller-Roeber B.
  TITLE     AssemblX: a user-friendly toolkit for rapid and reliable multi-gene 
            assemblies
  JOURNAL   Nucleic Acids Res. (2017) In press
  PUBMED    28130422
REFERENCE   2  (bases 1 to 5695)
  AUTHORS   Hochrein L, Machens F, Gremmels J, Schulz K, Messerschmidt K, 
            Mueller-Roeber B.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-NOV-2016) Department of Molecular Biology - Cell2Fab 
            research unit, University of Potsdam, Karl-Liebknecht-Str. 24-25, 
            Potsdam, Brandenburg 14476, Germany
REFERENCE   3  (bases 1 to 5695)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5695)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. (2017) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-NOV-2016) Department of Molecular Biology - Cell2Fab research 
            unit, University of Potsdam, Karl-Liebknecht-Str. 24-25, Potsdam, 
            Brandenburg 14476, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(568..1368)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(1369..1589)
                     /label=URA3 promoter
     rep_origin      complement(2188..3068)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3417..4208)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHDDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4209..4313)
                     /label=AmpR promoter
     misc_feature    4382..4421
                     /label=8 cutter set: AscI, SbfI, SwaI, FseI, PmeI
                     /note="8 cutter set: AscI, SbfI, SwaI, FseI, PmeI"
     misc_feature    4422..4471
                     /label=homology region E0*
                     /note="homology region E0*"
     CDS             4807..5109
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     promoter        5147..5427
                     /label=TRP1 promoter
     misc_feature    5428..5478
                     /label=homology region ER*
                     /note="homology region ER*"
     misc_feature    5482..5521
                     /label=8 cutter set: SwaI, AscI, PmeI, SbfI, FseI
                     /note="8 cutter set: SwaI, AscI, PmeI, SbfI, FseI"
     rep_origin      5547..5695
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
       61 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
      121 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
      181 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
      241 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
      301 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
      361 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
      421 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
      481 gctggccttt tgctcatgat ataattaaat tgaagctcta atttgtgagt ttagtataca
      541 tgcatttact tataatacag ttttttagtt ttgctggccg catcttctca aatatgcttc
      601 ccagcctgct tttctgtaac gttcaccctc taccttagca tcccttccct ttgcaaatag
      661 tcctcttcca acaataataa tgtcagatcc tgtagagacc acatcatcca cggttctata
      721 ctgttgaccc aatgcgtctc ccttgtcatc taaacccaca ccgggtgtca taatcaacca
      781 atcgtaacct tcatctcttc cacccatgtc tctttgagca ataaagccga taacaaaatc
      841 tttgtcgctc ttcgcaatgt caacagtacc cttagtatat tctccagtag atagggagcc
      901 cttgcatgac aattctgcta acatcaaaag gcctctaggt tcctttgtta cttcttctgc
      961 cgcctgcttc aaaccgctaa caatacctgg gcccaccaca ccgtgtgcat tcgtaatgtc
     1021 tgcccattct gctattctgt atacacccgc agagtactgc aatttgactg tattaccaat
     1081 gtcagcaaat tttctgtctt cgaagagtaa aaaattgtac ttggcggata atgcctttag
     1141 cggcttaact gtgccctcca tggaaaaatc agtcaagata tccacatgtg tttttagtaa
     1201 acaaattttg ggacctaatg cttcaactaa ctccagtaat tccttggtgg tacgaacatc
     1261 caatgaagca cacaagtttg tttgcttttc gtgcatgata ttaaatagct tggcagcaac
     1321 aggactagga tgagtagcag cacgttcctt atatgtagct ttcgacatga tttatcttcg
     1381 tttcctgcag gtttttgttc tgtgcagttg ggttaagaat actgggcaat ttcatgtttc
     1441 ttcaacacta catatgcgta tatataccaa tctaagtctg tgctccttcc ttcgttcttc
     1501 cttctgttcg gagattaccg aatcaaaaaa atttcaaaga aaccgaaatc aaaaaaaaga
     1561 ataaaaaaaa aatgatgaat tgaattgaaa agctagctta tcgatgataa gctgtcaaag
     1621 atgagaatta attccacgga ctatagacta tactagatac tccgtctact gtacgataca
     1681 cttccgctca ggtccttgtc ctttaacgag gccttaccac tcttttgtta ctctattgat
     1741 ccagctcagc aaaggcagtg tgatctaaga ttctatcttc gcgatgtagt aaaactagct
     1801 agaccgagaa agagactaga aatgcaaaag gcacttctac aatggctgcc atcattatta
     1861 tccgatgtga cgctgcagct tctcaatgat attcgaatac gctttgagga gatacagcct
     1921 aatatccgac aaactgtttt acagatttac gatcgtactt gttacccatc attgaatttt
     1981 gaacatccga acctgggagt tttccctgaa acagatagta tatttgaacc tgtataataa
     2041 tatatagtct agcgctttac ggaagacaat gtatgtattt cggttcctgg agaaactatt
     2101 gcatctattg cataggtaat cttgcacgtc gcatccccgg ttcattttct gcgtttccat
     2161 cttgcacttc aatagcatat ctttgttaac gaagcatctg tgcttcattt tgtagaacaa
     2221 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     2281 cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa
     2341 aacaaaaatg caacgcgacg agagcgctaa tttttcaaac aaagaatctg agctgcattt
     2401 ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc tatacttctt
     2461 ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca tcttagatta
     2521 ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt gcactgtagg
     2581 tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata aaaaaagcct
     2641 gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt tttcaagata
     2701 aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt gaacagaaag
     2761 tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt ctattttgtc
     2821 tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt cactctatga
     2881 atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac ataaaaaatg
     2941 tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag gttatatagg
     3001 gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt ggaagcggta
     3061 ttcgcaatgg gaagctccac cccggttgat aatcagaaaa gccccaaaaa caggaagatt
     3121 gtataagcaa atatttaaat tgtaattctt tcctgcgtta tcccctgatt ctgtggataa
     3181 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3241 cgagtcagtg agcgaggaag cggaagaggg tctgacgctc agtggaacga aaactcacgt
     3301 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3361 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagtcagaag
     3421 aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa
     3481 agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc
     3541 aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa
     3601 aagcggccat tttccaccat gatattcggc aagcaggcat cgtcatgggt cacgacgaga
     3661 tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc
     3721 tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct
     3781 cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc
     3841 agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac
     3901 aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca
     3961 acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc
     4021 tcgtcttgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc
     4081 ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag
     4141 tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt
     4201 tcaatcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4261 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4321 ccccgaaaag tgccacctga cgtcggacgg atcgcttgcc tgtaacttac acgcgcctcg
     4381 tggcgcgccc ctgcaggatt taaatggccg gccgtttaaa ctctcttaca ataaaataga
     4441 atactctgcg aagggcgtga cactgctgtt taagcttggc ttactaaaag ccagataaca
     4501 gtatgcgtat ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg
     4561 aagtatgtca aaaagaggta tgctatgaag cagcgtatta cagtgacagt tgacagcgac
     4621 agctatcagt tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca
     4681 tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga
     4741 tggctgaggt cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct
     4801 ggtgaaatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg
     4861 gatgtacaga gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt
     4921 gcacgtctgc tgtcagataa agtcccccgt gaactttacc cggtggtgca tatcggggat
     4981 gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa
     5041 gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc
     5101 tggggaatat aaatgtcagg ctcccttata cacagccagt aagcttaacg acattactat
     5161 atatataata taggaagcat ttaatagaca gcatcgtaat atatgtgtac tttgcagtta
     5221 tgacgccaga tggcagtagt ggaagatatt ctttattgaa aaatagcttg tcaccttacg
     5281 tacaatcttg atccggagct tttctttttt tgccgattaa gaattaattc ggtcgaaaaa
     5341 agaaaaggag agggccaaga gggagggcat tggtgactat tgagcacgtg agtatacgtg
     5401 attaagcaca caaaggcagc ttggagtatg tctgttatta atttcacagg tagttctggt
     5461 ccattggtga aagtttgctg aatttaaatg gcgcgccgtt taaaccctgc aggggccggc
     5521 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5581 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5641 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttc
//
