LOCUS       40924_27404        5382 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pL0E_3-4, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5382)
  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 5382)
  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 5382)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5382)
  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..5382
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(571..1371)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(1372..1592)
                     /label=URA3 promoter
     rep_origin      complement(2191..3071)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3420..4211)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHDDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4212..4316)
                     /label=AmpR promoter
     misc_feature    4385..4424
                     /label=8 cutter set: AscI, SbfI, SwaI, FseI, PmeI
                     /note="8 cutter set: AscI, SbfI, SwaI, FseI, PmeI"
     misc_feature    4425..4460
                     /label=homology region E3*
                     /note="homology region E3*"
     CDS             4796..5098
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     misc_feature    5136..5171
                     /label=homology region E4*
                     /note="homology region E4*"
     misc_feature    5172..5211
                     /label=8 cutter set: SwaI, AscI, PmeI, SbfI, FseI
                     /note="8 cutter set: SwaI, AscI, PmeI, SbfI, FseI"
     rep_origin      5237..5382
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
       61 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
      121 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
      181 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
      241 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
      301 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
      361 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
      421 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
      481 tttgctggcc ttttgctcat gatataatta aattgaagct ctaatttgtg agtttagtat
      541 acatgcattt acttataata cagtttttta gttttgctgg ccgcatcttc tcaaatatgc
      601 ttcccagcct gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa
      661 tagtcctctt ccaacaataa taatgtcaga tcctgtagag accacatcat ccacggttct
      721 atactgttga cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg tcataatcaa
      781 ccaatcgtaa ccttcatctc ttccacccat gtctctttga gcaataaagc cgataacaaa
      841 atctttgtcg ctcttcgcaa tgtcaacagt acccttagta tattctccag tagataggga
      901 gcccttgcat gacaattctg ctaacatcaa aaggcctcta ggttcctttg ttacttcttc
      961 tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg cattcgtaat
     1021 gtctgcccat tctgctattc tgtatacacc cgcagagtac tgcaatttga ctgtattacc
     1081 aatgtcagca aattttctgt cttcgaagag taaaaaattg tacttggcgg ataatgcctt
     1141 tagcggctta actgtgccct ccatggaaaa atcagtcaag atatccacat gtgtttttag
     1201 taaacaaatt ttgggaccta atgcttcaac taactccagt aattccttgg tggtacgaac
     1261 atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg atattaaata gcttggcagc
     1321 aacaggacta ggatgagtag cagcacgttc cttatatgta gctttcgaca tgatttatct
     1381 tcgtttcctg caggtttttg ttctgtgcag ttgggttaag aatactgggc aatttcatgt
     1441 ttcttcaaca ctacatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc
     1501 ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa agaaaccgaa atcaaaaaaa
     1561 agaataaaaa aaaaatgatg aattgaattg aaaagctagc ttatcgatga taagctgtca
     1621 aagatgagaa ttaattccac ggactataga ctatactaga tactccgtct actgtacgat
     1681 acacttccgc tcaggtcctt gtcctttaac gaggccttac cactcttttg ttactctatt
     1741 gatccagctc agcaaaggca gtgtgatcta agattctatc ttcgcgatgt agtaaaacta
     1801 gctagaccga gaaagagact agaaatgcaa aaggcacttc tacaatggct gccatcatta
     1861 ttatccgatg tgacgctgca gcttctcaat gatattcgaa tacgctttga ggagatacag
     1921 cctaatatcc gacaaactgt tttacagatt tacgatcgta cttgttaccc atcattgaat
     1981 tttgaacatc cgaacctggg agttttccct gaaacagata gtatatttga acctgtataa
     2041 taatatatag tctagcgctt tacggaagac aatgtatgta tttcggttcc tggagaaact
     2101 attgcatcta ttgcataggt aatcttgcac gtcgcatccc cggttcattt tctgcgtttc
     2161 catcttgcac ttcaatagca tatctttgtt aacgaagcat ctgtgcttca ttttgtagaa
     2221 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     2281 gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc ttcatttttg
     2341 taaaacaaaa atgcaacgcg acgagagcgc taatttttca aacaaagaat ctgagctgca
     2401 tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga atctatactt
     2461 cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa gcatcttaga
     2521 ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt tttgcactgt
     2581 aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc ataaaaaaag
     2641 cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca ttttttcaag
     2701 ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt tgtgaacaga
     2761 aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt cttctatttt
     2821 gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg attcactcta
     2881 tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata aacataaaaa
     2941 atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg taggttatat
     3001 agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt tgtggaagcg
     3061 gtattcgcaa tgggaagctc caccccggtt gataatcaga aaagccccaa aaacaggaag
     3121 attgtataag caaatattta aattgtaatt ctttcctgcg ttatcccctg attctgtgga
     3181 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3241 cagcgagtca gtgagcgagg aagcggaaga gggtctgacg ctcagtggaa cgaaaactca
     3301 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3361 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagtcag
     3421 aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg
     3481 taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta
     3541 gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca
     3601 gaaaagcggc cattttccac catgatattc ggcaagcagg catcgtcatg ggtcacgacg
     3661 agatcctcgc cgtcgggcat gctcgccttg agcctggcga acagttcggc tggcgcgagc
     3721 ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt
     3781 gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta
     3841 tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat
     3901 gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg
     3961 acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct
     4021 gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg
     4081 cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc
     4141 cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct
     4201 tgttcaatca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4261 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4321 tttccccgaa aagtgccacc tgacgtcgga cggatcgctt gcctgtaact tacacgcgcc
     4381 tcgtggcgcg cccctgcagg atttaaatgg ccggccgttt aaactaaaag taaactctca
     4441 cccgtaggat tggtatctgt aagcttggct tactaaaagc cagataacag tatgcgtatt
     4501 tgcgcgctga tttttgcggt ataagaatat atactgatat gtatacccga agtatgtcaa
     4561 aaagaggtat gctatgaagc agcgtattac agtgacagtt gacagcgaca gctatcagtt
     4621 gctcaaggca tatatgatgt caatatctcc ggtctggtaa gcacaaccat gcagaatgaa
     4681 gcccgtcgtc tgcgtgccga acgctggaaa gcggaaaatc aggaagggat ggctgaggtc
     4741 gcccggttta ttgaaatgaa cggctctttt gctgacgaga acaggggctg gtgaaatgca
     4801 gtttaaggtt tacacctata aaagagagag ccgttatcgt ctgtttgtgg atgtacagag
     4861 tgatattatt gacacgcccg ggcgacggat ggtgatcccc ctggccagtg cacgtctgct
     4921 gtcagataaa gtcccccgtg aactttaccc ggtggtgcat atcggggatg aaagctggcg
     4981 catgatgacc accgatatgg ccagtgtgcc ggtctccgtt atcggggaag aagtggctga
     5041 tctcagccac cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct ggggaatata
     5101 aatgtcaggc tcccttatac acagccagta agctttagtg atagcgttca atacttcctg
     5161 gcacgataat aatttaaatg gcgcgccgtt taaaccctgc aggggccggc cccgtagaaa
     5221 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5281 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5341 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tg
//
