LOCUS       40924_27429        5851 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pL1B-hc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5851)
  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 5851)
  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 5851)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5851)
  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..5851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(61..649)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(823..1680)
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      complement(2364..3244)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        3496..3764
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA
                     gene SNR52"
     misc_RNA        3773..3847
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      3852..3871
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4
                     tRNA gene"
     terminator      3923..4112
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     misc_feature    4331..4348
                     /note="I-SceI; cleavage site"
     misc_feature    4349..4398
                     /label=homology region B0*
                     /note="homology region B0*"
     CDS             4407..5498
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
     misc_feature    5603..5652
                     /label=homology region C0*
                     /note="homology region C0*"
     misc_feature    5653..5670
                     /note="I-SceI; cleavage site"
ORIGIN
        1 catgtgagca aaaggccagc aaaagcccag gaaccgtaaa aaggccgcgt tgctggcgtt
       61 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
      121 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      181 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      241 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      301 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      361 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      421 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      481 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
      541 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      601 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      661 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      721 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      781 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
      841 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
      901 gtagataact acgatacggg agcgcttacc atctggcccc agtgctgcaa tgataccgcg
      961 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     1021 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     1081 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttggca ttgctacagg
     1141 catcgtggtg tcactctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     1201 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1261 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1321 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     1381 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1441 ggataatagt gtatcacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     1501 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1561 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1621 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1681 actcttcctt tttcaatggg taataactga tataattaaa ttgaagctct aatttgtgag
     1741 tttagtatac atgcatttac ttataataca gttttatcga tgataagctg tcaaagatga
     1801 gaattaattc cacggactat agactatact agatactccg tctactgtac gatacacttc
     1861 cgctcaggtc cttgtccttt aacgaggcct taccactctt ttgttactct attgatccag
     1921 ctcagcaaag gcagtgtgat ctaagattct atcttcgcga tgtagtaaaa ctagctagac
     1981 cgagaaagag actagaaatg caaaaggcac ttctacaatg gctgccatca ttattatccg
     2041 atgtgacgct gcagcttctc aatgatattc gaatacgctt tgaggagata cagcctaata
     2101 tccgacaaac tgttttacag atttacgatc gtacttgtta cccatcattg aattttgaac
     2161 atccgaacct gggagttttc cctgaaacag atagtatatt tgaacctgta taataatata
     2221 tagtctagcg ctttacggaa gacaatgtat gtatttcggt tcctggagaa actattgcat
     2281 ctattgcata ggtaatcttg cacgtcgcat ccccggttca ttttctgcgt ttccatcttg
     2341 cacttcaata gcatatcttt gttaacgaag catctgtgct tcattttgta gaacaaaaat
     2401 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     2461 aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca
     2521 aaaatgcaac gcgacgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     2581 agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt
     2641 gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt
     2701 ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg
     2761 ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact
     2821 ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg
     2881 catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat
     2941 agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta
     3001 tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag
     3061 ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga
     3121 ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata
     3181 tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg
     3241 caatgacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3301 taacgaatag cccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     3361 gttgagtgtt gttccagttt ccaacaagag tccactatta aagaacgtgg actccaacgt
     3421 caaagggcga aaaagggtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     3481 aagttttttg gggtctcttt gaaaagataa tgtatgatta tgctttcact catatttata
     3541 cagaaacttg atgttttctt tcgagtatat acaaggtgat tacatgtacg tttgaagtac
     3601 aactctagat tttgtagtgc cctcttgggc tagcggtaaa ggtgcgcatt ttttcacacc
     3661 ctacaatgtt ctgttcaaaa gattttggtc aaacgctgta gaagtgaaag ttggtgcgca
     3721 tgtttcggcg ttcgaaactt ctccgcagtg aaagataaat gatcgcggcc gcgttttaga
     3781 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
     3841 gtcggtggtg ctttttttgt tttttatgtc ttcgagtcat gtaattagtt atgtcacgct
     3901 tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg
     3961 aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat
     4021 ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa
     4081 ccttgcttga gaaggttttg ggacgctcga aggctttagg tgccgtaaag cagtaaatcg
     4141 gaagggtaaa cggatgcccc catttagagc ttgacgggga aagccggcga acgtggcgag
     4201 aaaggaaggg aagaaagcga aaggagcggg ggctagggcg gtgggaagtg taggggtcac
     4261 gctgggcgta accaccacac ccgccgcgct taatggggcg ctacagggcg cgtggggatg
     4321 atccactagt tagggataac agggtaatta aggtttcagt cacactacgg atacttttac
     4381 aacgggagca gttattcatt aattaaatgt ctgcccctaa gaagatcgtc gttttgccag
     4441 gtgaccacgt tggtcaagaa atcacagccg aagccattaa ggttcttaaa gctatttctg
     4501 atgttcgttc caatgtcaag ttcgatttcg aaaatcattt aattggtggt gctgctatcg
     4561 atgctacagg tgtcccactt ccagatgagg cgctggaagc ctccaagaag gttgatgccg
     4621 ttttgttagg tgctgtgggt ggtcctaaat ggggtacagg tagtgttaga cctgaacaag
     4681 gtttactaaa aatccgtaaa gaacttcaat tgtacgccaa cttaagacca tgtaactttg
     4741 catccgactc tcttttagac ttatctccaa tcaagccaca atttgctaaa ggtactgact
     4801 tcgttgttgt cagagaatta gtgggaggta tttactttgg taagagaaag gaagacgatg
     4861 gtgatggtgt cgcttgggat agtgaacaat acaccgttcc agaagtgcaa agaatcacaa
     4921 gaatggccgc tttcatggcc ctacaacatg agccaccatt gcctatttgg tccttggata
     4981 aagctaatgt tttggcctct tcaagattat ggagaaaaac tgtggaggaa accatcaaga
     5041 acgaatttcc tacattgaag gttcaacatc aattgattga ttctgccgcc atgatcctag
     5101 ttaagaaccc aacccaccta aatggtatta taatcaccag caacatgttt ggtgatatca
     5161 tctccgatga agcctccgtt atcccaggtt ccttgggttt gttgccatct gcgtccttgg
     5221 cctctttgcc agacaagaac accgcatttg gtttgtacga accatgccac ggttctgctc
     5281 cagatttgcc aaagaataag gttgacccta tcgccactat cttgtctgct gcaatgatgt
     5341 tgaaattgtc attgaacttg cctgaagaag gtaaggccat tgaagatgca gttaaaaagg
     5401 ttttggatgc aggtatcaga actggtgatt taggtggttc caacagtacc accgaagtcg
     5461 gtgatgctgt cgccgaagaa gttaagaaaa tccttgctta aaaagattct ctttttttat
     5521 gatatttgta cataaacttt ataaatgaaa ttcataatag aaacgacacg aaattacaaa
     5581 atggaatatg ttcatagggt agtattgcgt gtgatacttc aggagaaccg ttattccgta
     5641 aaactacagt caattaccct gttatcccta cggccctgca ttaatgaatc ggccaacgcg
     5701 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5761 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5821 ccacagaatc aggggataac gcaggaaaga a
//
