LOCUS       40924_27504        4653 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pL2AD-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(60..648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(822..1679)
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      complement(2363..3243)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        3495..3763
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA
                     gene SNR52"
     misc_RNA        3772..3846
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      3851..3870
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4
                     tRNA gene"
     terminator      3922..4111
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     misc_feature    4330..4347
                     /note="I-SceI; cleavage site"
     misc_feature    4348..4397
                     /label=homology region A0*
                     /note="homology region A0*"
     misc_feature    4404..4453
                     /label=homology region E0*
                     /note="homology region E0*"
     misc_feature    4454..4471
                     /note="I-SceI; cleavage site"
ORIGIN
        1 atgtgagcaa aaggccagca aaagcccagg aaccgtaaaa aggccgcgtt gctggcgttt
       61 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
      121 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
      181 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
      241 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
      301 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
      361 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
      421 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
      481 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
      541 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
      601 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
      661 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
      721 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
      781 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
      841 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
      901 tagataacta cgatacggga gcgcttacca tctggcccca gtgctgcaat gataccgcga
      961 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     1021 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     1081 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttggcat tgctacaggc
     1141 atcgtggtgt cactctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     1201 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     1261 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     1321 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     1381 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     1441 gataatagtg tatcacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     1501 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     1561 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     1621 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     1681 ctcttccttt ttcaatgggt aataactgat ataattaaat tgaagctcta atttgtgagt
     1741 ttagtataca tgcatttact tataatacag ttttatcgat gataagctgt caaagatgag
     1801 aattaattcc acggactata gactatacta gatactccgt ctactgtacg atacacttcc
     1861 gctcaggtcc ttgtccttta acgaggcctt accactcttt tgttactcta ttgatccagc
     1921 tcagcaaagg cagtgtgatc taagattcta tcttcgcgat gtagtaaaac tagctagacc
     1981 gagaaagaga ctagaaatgc aaaaggcact tctacaatgg ctgccatcat tattatccga
     2041 tgtgacgctg cagcttctca atgatattcg aatacgcttt gaggagatac agcctaatat
     2101 ccgacaaact gttttacaga tttacgatcg tacttgttac ccatcattga attttgaaca
     2161 tccgaacctg ggagttttcc ctgaaacaga tagtatattt gaacctgtat aataatatat
     2221 agtctagcgc tttacggaag acaatgtatg tatttcggtt cctggagaaa ctattgcatc
     2281 tattgcatag gtaatcttgc acgtcgcatc cccggttcat tttctgcgtt tccatcttgc
     2341 acttcaatag catatctttg ttaacgaagc atctgtgctt cattttgtag aacaaaaatg
     2401 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     2461 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
     2521 aaatgcaacg cgacgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     2581 gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg
     2641 ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt
     2701 tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt
     2761 taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc
     2821 cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc
     2881 atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata
     2941 gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat
     3001 atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt
     3061 tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag
     3121 gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat
     3181 agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc
     3241 aatgacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3301 aacgaatagc ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3361 ttgagtgttg ttccagtttc caacaagagt ccactattaa agaacgtgga ctccaacgtc
     3421 aaagggcgaa aaagggtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     3481 agttttttgg ggtctctttg aaaagataat gtatgattat gctttcactc atatttatac
     3541 agaaacttga tgttttcttt cgagtatata caaggtgatt acatgtacgt ttgaagtaca
     3601 actctagatt ttgtagtgcc ctcttgggct agcggtaaag gtgcgcattt tttcacaccc
     3661 tacaatgttc tgttcaaaag attttggtca aacgctgtag aagtgaaagt tggtgcgcat
     3721 gtttcggcgt tcgaaacttc tccgcagtga aagataaatg atcgcggccg cgttttagag
     3781 ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
     3841 tcggtggtgc tttttttgtt ttttatgtct tcgagtcatg taattagtta tgtcacgctt
     3901 acattcacgc cctcccccca catccgctct aaccgaaaag gaaggagtta gacaacctga
     3961 agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt
     4021 tcaaattttt cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac
     4081 cttgcttgag aaggttttgg gacgctcgaa ggctttaggt gccgtaaagc agtaaatcgg
     4141 aagggtaaac ggatgccccc atttagagct tgacggggaa agccggcgaa cgtggcgaga
     4201 aaggaaggga agaaagcgaa aggagcgggg gctagggcgg tgggaagtgt aggggtcacg
     4261 ctgggcgtaa ccaccacacc cgccgcgctt aatggggcgc tacagggcgc gtggggatga
     4321 tccactagtt agggataaca gggtaataaa taggaaagtc tggtaactca ggacactgta
     4381 tctgctacgc tgtttatgaa ttctctctta caataaaata gaatactctg cgaagggcgt
     4441 gacactgctg tttattaccc tgttatccct acggccctgc attaatgaat cggccaacgc
     4501 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4561 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4621 tccacagaat caggggataa cgcaggaaag aac
//
