LOCUS       40924_27464        4472 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pL1E-lc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4472)
  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 4472)
  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 4472)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4472)
  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..4472
                     /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)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    complement(1781..2284)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        2545..2813
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA
                     gene SNR52"
     misc_RNA        2822..2896
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      2901..2920
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4
                     tRNA gene"
     terminator      2972..3161
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     misc_feature    3380..3397
                     /label=I-SceI
                     /note="I-SceI"
     misc_feature    3398..3447
                     /label=homology region E0*
                     /note="homology region E0*"
     CDS             3456..4127
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     misc_feature    4227..4276
                     /label=homology region F0*
                     /note="homology region F0*"
     misc_feature    4277..4294
                     /label=I-SceI
                     /note="I-SceI"
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 acagttgacg gatcgcttgc ctgtaactta
     1801 cacgcgcctc gtatctttta atgatggaat aatttgggaa tttactctgt gtttatttat
     1861 ttttatgttt tgtatttgga ttttagaaag taaataaaga aggtagaaga gttacggaat
     1921 gaagaaaaaa aaataaacaa aggtttaaaa aatttcaaca aaaagcgtac tttacatata
     1981 tatttattag acaagaaaag cagattaaat agatatacat tcgattaacg ataagtaaaa
     2041 tgtaaaatca caggattttc gtgtgtggtc ttctacacag acaagatgaa acaattcggc
     2101 attaatacct gagagcagga agagcaagat aaaaggtagt atttgttggc gatcccccta
     2161 gagtctttta catcttcgga aaacaaaaac tattttttct ttaatttctt tttttacttt
     2221 ctatttttaa tttatatatt tatattaaaa aatttaaatt ataattattt ttatagcacg
     2281 tgatgaaaag gaccacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     2341 ctcatttttt aacgaatagc ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     2401 cgagataggg ttgagtgttg ttccagtttc caacaagagt ccactattaa agaacgtgga
     2461 ctccaacgtc aaagggcgaa aaagggtcta tcagggcgat ggcccactac gtgaaccatc
     2521 accctaatca agttttttgg ggtctctttg aaaagataat gtatgattat gctttcactc
     2581 atatttatac agaaacttga tgttttcttt cgagtatata caaggtgatt acatgtacgt
     2641 ttgaagtaca actctagatt ttgtagtgcc ctcttgggct agcggtaaag gtgcgcattt
     2701 tttcacaccc tacaatgttc tgttcaaaag attttggtca aacgctgtag aagtgaaagt
     2761 tggtgcgcat gtttcggcgt tcgaaacttc tccgcagtga aagataaatg atcgcggccg
     2821 cgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag
     2881 tggcaccgag tcggtggtgc tttttttgtt ttttatgtct tcgagtcatg taattagtta
     2941 tgtcacgctt acattcacgc cctcccccca catccgctct aaccgaaaag gaaggagtta
     3001 gacaacctga agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt
     3061 tatttatatt tcaaattttt cttttttttc tgtacagacg cgtgtacgca tgtaacatta
     3121 tactgaaaac cttgcttgag aaggttttgg gacgctcgaa ggctttaggt gccgtaaagc
     3181 agtaaatcgg aagggtaaac ggatgccccc atttagagct tgacggggaa agccggcgaa
     3241 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggg gctagggcgg tgggaagtgt
     3301 aggggtcacg ctgggcgtaa ccaccacacc cgccgcgctt aatggggcgc tacagggcgc
     3361 gtggggatga tccactagtt agggataaca gggtaattct cttacaataa aatagaatac
     3421 tctgcgaagg gcgtgacact gctgttttta attaaatgtc tgttattaat ttcacaggta
     3481 gttctggtcc attggtgaaa gtttgcggct tgcagagcac agaggccgca gaatgtgctc
     3541 tagattccga tgctgacttg ctgggtatta tatgtgtgcc caatagaaag agaacaattg
     3601 acccggttat tgcaaggaaa atttcaagtc ttgtaaaagc atataaaaat agttcaggca
     3661 ctccgaaata cttggttggc gtgtttcgta atcaacctaa ggaggatgtt ttggctctgg
     3721 tcaatgatta cggcattgat atcgtccaac tgcatggaga tgagtcgtgg caagaatacc
     3781 aagagttcct cggtttgcca gttattaaaa gactcgtatt tccaaaagac tgcaacatac
     3841 tactcagtgc agcttcacag aaacctcatt cgtttattcc cttgtttgat tcagaagcag
     3901 gtgggacagg tgaacttttg gattggaact cgatttctga ctgggttgga aggcaagaga
     3961 gccccgaaag cttacatttt atgttagctg gtggactgac gccagaaaat gttggtgatg
     4021 cgcttagatt aaatggcgtt attggtgttg atgtaagcgg aggtgtggag acaaatggtg
     4081 taaaagactc taacaaaata gcaaatttcg tcaaaaatgc taagaaatag gttattactg
     4141 agtagtattt atttaagtat tgtttgtgca cttgcctatg cggtgtgaaa taccgcacag
     4201 atgcgtaagg agaaaatacc gcatcaaaag tattggtgtg agtaatcgct atctcttcta
     4261 cgactggcta caacaaatta ccctgttatc cctacggccc tgcattaatg aatcggccaa
     4321 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4381 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4441 ttatccacag aatcagggga taacgcagga aa
//
