LOCUS       40924_34161        6280 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPC_001_GFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6280)
  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 6280)
  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 6280)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6280)
  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..6280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(470..1270)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(1271..1491)
                     /label=URA3 promoter
     rep_origin      complement(2090..2970)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3319..4110)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHDDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4111..4215)
                     /label=AmpR promoter
     promoter        4371..4767
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             4782..5495
                     /codon_start=1
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      5507..5672
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     promoter        5693..5913
                     /label=URA3 promoter
     rep_origin      6034..6280
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
       61 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
      121 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
      181 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
      241 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
      301 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
      361 tttttacggt tcctggcctt ttgctggcct tttgctcatg atataattaa attgaagctc
      421 taatttgtga gtttagtata catgcattta cttataatac agttttttag ttttgctggc
      481 cgcatcttct caaatatgct tcccagcctg cttttctgta acgttcaccc tctaccttag
      541 catcccttcc ctttgcaaat agtcctcttc caacaataat aatgtcagat cctgtagaga
      601 ccacatcatc cacggttcta tactgttgac ccaatgcgtc tcccttgtca tctaaaccca
      661 caccgggtgt cataatcaac caatcgtaac cttcatctct tccacccatg tctctttgag
      721 caataaagcc gataacaaaa tctttgtcgc tcttcgcaat gtcaacagta cccttagtat
      781 attctccagt agatagggag cccttgcatg acaattctgc taacatcaaa aggcctctag
      841 gttcctttgt tacttcttct gccgcctgct tcaaaccgct aacaatacct gggcccacca
      901 caccgtgtgc attcgtaatg tctgcccatt ctgctattct gtatacaccc gcagagtact
      961 gcaatttgac tgtattacca atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt
     1021 acttggcgga taatgccttt agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga
     1081 tatccacatg tgtttttagt aaacaaattt tgggacctaa tgcttcaact aactccagta
     1141 attccttggt ggtacgaaca tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga
     1201 tattaaatag cttggcagca acaggactag gatgagtagc agcacgttcc ttatatgtag
     1261 ctttcgacat gatttatctt cgtttcctgc aggtttttgt tctgtgcagt tgggttaaga
     1321 atactgggca atttcatgtt tcttcaacac tacatatgcg tatatatacc aatctaagtc
     1381 tgtgctcctt ccttcgttct tccttctgtt cggagattac cgaatcaaaa aaatttcaaa
     1441 gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga attgaattga aaagctagct
     1501 tatcgatgat aagctgtcaa agatgagaat taattccacg gactatagac tatactagat
     1561 actccgtcta ctgtacgata cacttccgct caggtccttg tcctttaacg aggccttacc
     1621 actcttttgt tactctattg atccagctca gcaaaggcag tgtgatctaa gattctatct
     1681 tcgcgatgta gtaaaactag ctagaccgag aaagagacta gaaatgcaaa aggcacttct
     1741 acaatggctg ccatcattat tatccgatgt gacgctgcag cttctcaatg atattcgaat
     1801 acgctttgag gagatacagc ctaatatccg acaaactgtt ttacagattt acgatcgtac
     1861 ttgttaccca tcattgaatt ttgaacatcc gaacctggga gttttccctg aaacagatag
     1921 tatatttgaa cctgtataat aatatatagt ctagcgcttt acggaagaca atgtatgtat
     1981 ttcggttcct ggagaaacta ttgcatctat tgcataggta atcttgcacg tcgcatcccc
     2041 ggttcatttt ctgcgtttcc atcttgcact tcaatagcat atctttgtta acgaagcatc
     2101 tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa
     2161 tctgagctgc atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag
     2221 aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga cgagagcgct aatttttcaa
     2281 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac
     2341 caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt
     2401 tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct
     2461 tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt
     2521 ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct
     2581 gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg
     2641 cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat
     2701 gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta
     2761 ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata
     2821 ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag
     2881 gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga gatacttttg
     2941 agcaatgttt gtggaagcgg tattcgcaat gggaagctcc accccggttg ataatcagaa
     3001 aagccccaaa aacaggaaga ttgtataagc aaatatttaa attgtaattc tttcctgcgt
     3061 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3121 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag ggtctgacgc
     3181 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3241 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3301 aacttggtct gacagtcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga
     3361 atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc
     3421 ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg
     3481 gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc
     3541 atcgtcatgg gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa
     3601 cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc
     3661 ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca
     3721 ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc
     3781 ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca
     3841 gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc
     3901 cagccacgat agccgcgctg cctcgtcttg cagttcattc agggcaccgg acaggtcggt
     3961 cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca
     4021 gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga
     4081 acctgcgtgc aatccatctt gttcaatcat actcttcctt tttcaatatt attgaagcat
     4141 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4201 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gcggacggat cgcttgcctg
     4261 taacttacac gcgcctcgtg gcgcgcccct gcaggattta aatggccggc cgtttaaacc
     4321 ttgtacacct accggtcacc agccaacaat gtgcggatgg cgttacaact caacttcttt
     4381 tctttttttt tcttttctct ctcccccgtt gttgtctcac catatccgca atgacaaaaa
     4441 aatgatggaa gacactaaag gaaaaaatta acgacaaaga cagcaccaac agatgtcgtt
     4501 gttccagagc tgatgagggg tatctcgaag cacacgaaac tttttccttc cttcattcac
     4561 gcacactact ctctaatgag caacggtata cggccttcct tccagttact tgaatttgaa
     4621 ataaaaaaaa gtttgctgtc ttgctatcaa gtataaatag acctgcaatt attaatcttt
     4681 tgtttcctcg tcattgttct cgttcccttt cttccttgtt tctttttctg cacaatattt
     4741 caagctatac caagcataca atcaactatc tcagcggccg catgtctaaa ggtgaagaat
     4801 tattcactgg tgttgtccca attttggttg aattagatgg tgatgttaat ggtcacaaat
     4861 tttctgtctc cggtgaaggt gaaggtgatg ctacttacgg taaattgacc ttaaaattta
     4921 tttgtactac tggtaaattg ccagttccat ggccaacctt agtcactact ttcggttatg
     4981 gtgttcaatg ttttgctaga tacccagatc atatgaaaca acatgacttt ttcaagtctg
     5041 ccatgccaga aggttatgtt caagaaagaa ctattttttt caaagatgac ggtaactaca
     5101 agaccagagc tgaagtcaag tttgaaggtg ataccttagt taatagaatc gaattaaaag
     5161 gtattgattt taaagaagat ggtaacattt taggtcacaa attggaatac aactataact
     5221 ctcacaatgt ttacatcatg gctgacaaac aaaagaatgg tatcaaagtt aacttcaaaa
     5281 ttagacacaa cattgaagat ggttctgttc aattagctga ccattatcaa caaaatactc
     5341 caattggtga tggtccagtc ttgttaccag acaaccatta cttatccact caatctgcct
     5401 tatccaaaga tccaaacgaa aagagagacc acatggtctt gttagaattt gttactgctg
     5461 ctggtattac ccatggtatg gatgaattgt acaaataagc ggccgcgcga atttcttatg
     5521 atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca aattttaaag
     5581 tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc tgtaggtcag
     5641 gttgctttct caggtatagc atgaggtcgc tcttattgac cacacctagc ttttcaattc
     5701 aattcatcat ttttttttta ttcttttttt tgatttcggt ttctttgaaa tttttttgat
     5761 tcggtaatct ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat
     5821 atacgcatat gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac
     5881 agaacaaaaa cctgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt
     5941 gctgctactc atcctagtcc tgtttgagat ttaaatggcg cgccgtttaa accctgcagg
     6001 ggccggcccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6061 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6121 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6181 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6241 cctcgctctg ctaatcctgt taccagtggc tgctgccagt
//
