LOCUS       40924_27514        4653 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pL2AE-hc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4653)
  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 4653)
  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 4653)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4653)
  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..4653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(57..645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(819..1676)
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      complement(2360..3240)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        3492..3760
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA
                     gene SNR52"
     misc_RNA        3769..3843
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      3848..3867
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4
                     tRNA gene"
     terminator      3919..4108
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     misc_feature    4327..4344
                     /note="I-SceI; cleavage site"
     misc_feature    4345..4394
                     /label=homology region A0*
                     /note="homology region A0*"
     misc_feature    4401..4450
                     /label=homology region F0*
                     /note="homology region F0*"
     misc_feature    4451..4468
                     /note="I-SceI; cleavage site"
ORIGIN
        1 tgagcaaaag gccagcaaaa gcccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
       61 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      121 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      181 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      241 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      301 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      361 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      421 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      481 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      541 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      601 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      661 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      721 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
      781 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
      841 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
      901 ataactacga tacgggagcg cttaccatct ggccccagtg ctgcaatgat accgcgagac
      961 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1021 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     1081 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttggcattgc tacaggcatc
     1141 gtggtgtcac tctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1201 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1261 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1321 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1381 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1441 aatagtgtat cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1501 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1561 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1621 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     1681 ttcctttttc aatgggtaat aactgatata attaaattga agctctaatt tgtgagttta
     1741 gtatacatgc atttacttat aatacagttt tatcgatgat aagctgtcaa agatgagaat
     1801 taattccacg gactatagac tatactagat actccgtcta ctgtacgata cacttccgct
     1861 caggtccttg tcctttaacg aggccttacc actcttttgt tactctattg atccagctca
     1921 gcaaaggcag tgtgatctaa gattctatct tcgcgatgta gtaaaactag ctagaccgag
     1981 aaagagacta gaaatgcaaa aggcacttct acaatggctg ccatcattat tatccgatgt
     2041 gacgctgcag cttctcaatg atattcgaat acgctttgag gagatacagc ctaatatccg
     2101 acaaactgtt ttacagattt acgatcgtac ttgttaccca tcattgaatt ttgaacatcc
     2161 gaacctggga gttttccctg aaacagatag tatatttgaa cctgtataat aatatatagt
     2221 ctagcgcttt acggaagaca atgtatgtat ttcggttcct ggagaaacta ttgcatctat
     2281 tgcataggta atcttgcacg tcgcatcccc ggttcatttt ctgcgtttcc atcttgcact
     2341 tcaatagcat atctttgtta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa
     2401 cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg
     2461 caacgcgaaa gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa
     2521 tgcaacgcga cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     2581 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     2641 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     2701 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     2761 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     2821 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     2881 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     2941 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     3001 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     3061 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     3121 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     3181 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     3241 gacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3301 gaatagcccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3361 agtgttgttc cagtttccaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3421 gggcgaaaaa gggtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3481 tttttggggt ctctttgaaa agataatgta tgattatgct ttcactcata tttatacaga
     3541 aacttgatgt tttctttcga gtatatacaa ggtgattaca tgtacgtttg aagtacaact
     3601 ctagattttg tagtgccctc ttgggctagc ggtaaaggtg cgcatttttt cacaccctac
     3661 aatgttctgt tcaaaagatt ttggtcaaac gctgtagaag tgaaagttgg tgcgcatgtt
     3721 tcggcgttcg aaacttctcc gcagtgaaag ataaatgatc gcggccgcgt tttagagcta
     3781 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     3841 gtggtgcttt ttttgttttt tatgtcttcg agtcatgtaa ttagttatgt cacgcttaca
     3901 ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt
     3961 ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat ttatatttca
     4021 aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt
     4081 gcttgagaag gttttgggac gctcgaaggc tttaggtgcc gtaaagcagt aaatcggaag
     4141 ggtaaacgga tgcccccatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     4201 gaagggaaga aagcgaaagg agcgggggct agggcggtgg gaagtgtagg ggtcacgctg
     4261 ggcgtaacca ccacacccgc cgcgcttaat ggggcgctac agggcgcgtg gggatgatcc
     4321 actagttagg gataacaggg taataaatag gaaagtctgg taactcagga cactgtatct
     4381 gctacgctgt ttatgaattc aaagtattgg tgtgagtaat cgctatctct tctacgactg
     4441 gctacaacaa attaccctgt tatccctacg gccctgcatt aatgaatcgg ccaacgcgcg
     4501 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4561 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4621 acagaatcag gggataacgc aggaaagaac atg
//
