LOCUS       40924_27459        5426 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pL1E-hc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5426)
  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 5426)
  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 5426)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5426)
  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..5426
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(64..652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(826..1683)
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      complement(2367..3247)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        3499..3767
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA
                     gene SNR52"
     misc_RNA        3776..3850
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      3855..3874
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4
                     tRNA gene"
     terminator      3926..4115
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     misc_feature    4334..4351
                     /note="I-SceI; cleavage site"
     misc_feature    4352..4401
                     /label=homology region E0*
                     /note="homology region E0*"
     CDS             4410..5081
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
     misc_feature    5181..5230
                     /label=homology region F0*
                     /note="homology region F0*"
     misc_feature    5231..5248
                     /note="I-SceI; cleavage site"
ORIGIN
        1 gaacatgtga gcaaaaggcc agcaaaagcc caggaaccgt aaaaaggccg cgttgctggc
       61 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      121 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      181 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      241 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      301 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      361 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      421 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      481 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
      541 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      601 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      661 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      721 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      781 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
      841 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
      901 cgtgtagata actacgatac gggagcgctt accatctggc cccagtgctg caatgatacc
      961 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1021 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1081 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg gcattgctac
     1141 aggcatcgtg gtgtcactct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1201 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1261 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1321 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1381 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1441 acgggataat agtgtatcac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1501 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1561 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1621 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1681 catactcttc ctttttcaat gggtaataac tgatataatt aaattgaagc tctaatttgt
     1741 gagtttagta tacatgcatt tacttataat acagttttat cgatgataag ctgtcaaaga
     1801 tgagaattaa ttccacggac tatagactat actagatact ccgtctactg tacgatacac
     1861 ttccgctcag gtccttgtcc tttaacgagg ccttaccact cttttgttac tctattgatc
     1921 cagctcagca aaggcagtgt gatctaagat tctatcttcg cgatgtagta aaactagcta
     1981 gaccgagaaa gagactagaa atgcaaaagg cacttctaca atggctgcca tcattattat
     2041 ccgatgtgac gctgcagctt ctcaatgata ttcgaatacg ctttgaggag atacagccta
     2101 atatccgaca aactgtttta cagatttacg atcgtacttg ttacccatca ttgaattttg
     2161 aacatccgaa cctgggagtt ttccctgaaa cagatagtat atttgaacct gtataataat
     2221 atatagtcta gcgctttacg gaagacaatg tatgtatttc ggttcctgga gaaactattg
     2281 catctattgc ataggtaatc ttgcacgtcg catccccggt tcattttctg cgtttccatc
     2341 ttgcacttca atagcatatc tttgttaacg aagcatctgt gcttcatttt gtagaacaaa
     2401 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
     2461 agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa
     2521 acaaaaatgc aacgcgacga gagcgctaat ttttcaaaca aagaatctga gctgcatttt
     2581 tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct atacttcttt
     2641 tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat cttagattac
     2701 tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg cactgtaggt
     2761 ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa aaaaagcctg
     2821 actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt ttcaagataa
     2881 aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg aacagaaagt
     2941 gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc tattttgtct
     3001 ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc actctatgaa
     3061 tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca taaaaaatgt
     3121 agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg ttatataggg
     3181 atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg gaagcggtat
     3241 tcgcaatgac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3301 ttttaacgaa tagcccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3361 agggttgagt gttgttccag tttccaacaa gagtccacta ttaaagaacg tggactccaa
     3421 cgtcaaaggg cgaaaaaggg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3481 atcaagtttt ttggggtctc tttgaaaaga taatgtatga ttatgctttc actcatattt
     3541 atacagaaac ttgatgtttt ctttcgagta tatacaaggt gattacatgt acgtttgaag
     3601 tacaactcta gattttgtag tgccctcttg ggctagcggt aaaggtgcgc attttttcac
     3661 accctacaat gttctgttca aaagattttg gtcaaacgct gtagaagtga aagttggtgc
     3721 gcatgtttcg gcgttcgaaa cttctccgca gtgaaagata aatgatcgcg gccgcgtttt
     3781 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac
     3841 cgagtcggtg gtgctttttt tgttttttat gtcttcgagt catgtaatta gttatgtcac
     3901 gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac
     3961 ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta
     4021 tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga
     4081 aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aggtgccgta aagcagtaaa
     4141 tcggaagggt aaacggatgc ccccatttag agcttgacgg ggaaagccgg cgaacgtggc
     4201 gagaaaggaa gggaagaaag cgaaaggagc gggggctagg gcggtgggaa gtgtaggggt
     4261 cacgctgggc gtaaccacca cacccgccgc gcttaatggg gcgctacagg gcgcgtgggg
     4321 atgatccact agttagggat aacagggtaa ttctcttaca ataaaataga atactctgcg
     4381 aagggcgtga cactgctgtt tttaattaaa tgtctgttat taatttcaca ggtagttctg
     4441 gtccattggt gaaagtttgc ggcttgcaga gcacagaggc cgcagaatgt gctctagatt
     4501 ccgatgctga cttgctgggt attatatgtg tgcccaatag aaagagaaca attgacccgg
     4561 ttattgcaag gaaaatttca agtcttgtaa aagcatataa aaatagttca ggcactccga
     4621 aatacttggt tggcgtgttt cgtaatcaac ctaaggagga tgttttggct ctggtcaatg
     4681 attacggcat tgatatcgtc caactgcatg gagatgagtc gtggcaagaa taccaagagt
     4741 tcctcggttt gccagttatt aaaagactcg tatttccaaa agactgcaac atactactca
     4801 gtgcagcttc acagaaacct cattcgttta ttcccttgtt tgattcagaa gcaggtggga
     4861 caggtgaact tttggattgg aactcgattt ctgactgggt tggaaggcaa gagagccccg
     4921 aaagcttaca ttttatgtta gctggtggac tgacgccaga aaatgttggt gatgcgctta
     4981 gattaaatgg cgttattggt gttgatgtaa gcggaggtgt ggagacaaat ggtgtaaaag
     5041 actctaacaa aatagcaaat ttcgtcaaaa atgctaagaa ataggttatt actgagtagt
     5101 atttatttaa gtattgtttg tgcacttgcc tatgcggtgt gaaataccgc acagatgcgt
     5161 aaggagaaaa taccgcatca aaagtattgg tgtgagtaat cgctatctct tctacgactg
     5221 gctacaacaa attaccctgt tatccctacg gccctgcatt aatgaatcgg ccaacgcgcg
     5281 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5341 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5401 acagaatcag gggataacgc aggaaa
//
