LOCUS       40924_34356        6427 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPC_040_GFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6427)
  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 6427)
  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 6427)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6427)
  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..6427
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(469..1269)
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(1270..1490)
                     /label=URA3 promoter
     rep_origin      complement(2089..2969)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3318..4109)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(4110..4214)
                     /label=AmpR promoter
     regulatory      4370..4776
                     /label=ZEO1 promoter
                     /note="ZEO1 promoter"
                     /regulatory_class="promoter"
     CDS             4785..5498
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
     RBS             5569..5580
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             complement(5716..5736)
                     /label=7xHis
                     /note="6xHis affinity tag"
     promoter        5839..6059
                     /label=URA3 promoter
     rep_origin      6180..6427
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
       61 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
      121 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
      181 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
      241 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
      301 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
      361 ttttacggtt cctggccttt tgctggcctt ttgctcatga tataattaaa ttgaagctct
      421 aatttgtgag tttagtatac atgcatttac ttataataca gttttttagt tttgctggcc
      481 gcatcttctc aaatatgctt cccagcctgc ttttctgtaa cgttcaccct ctaccttagc
      541 atcccttccc tttgcaaata gtcctcttcc aacaataata atgtcagatc ctgtagagac
      601 cacatcatcc acggttctat actgttgacc caatgcgtct cccttgtcat ctaaacccac
      661 accgggtgtc ataatcaacc aatcgtaacc ttcatctctt ccacccatgt ctctttgagc
      721 aataaagccg ataacaaaat ctttgtcgct cttcgcaatg tcaacagtac ccttagtata
      781 ttctccagta gatagggagc ccttgcatga caattctgct aacatcaaaa ggcctctagg
      841 ttcctttgtt acttcttctg ccgcctgctt caaaccgcta acaatacctg ggcccaccac
      901 accgtgtgca ttcgtaatgt ctgcccattc tgctattctg tatacacccg cagagtactg
      961 caatttgact gtattaccaa tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta
     1021 cttggcggat aatgccttta gcggcttaac tgtgccctcc atggaaaaat cagtcaagat
     1081 atccacatgt gtttttagta aacaaatttt gggacctaat gcttcaacta actccagtaa
     1141 ttccttggtg gtacgaacat ccaatgaagc acacaagttt gtttgctttt cgtgcatgat
     1201 attaaatagc ttggcagcaa caggactagg atgagtagca gcacgttcct tatatgtagc
     1261 tttcgacatg atttatcttc gtttcctgca ggtttttgtt ctgtgcagtt gggttaagaa
     1321 tactgggcaa tttcatgttt cttcaacact acatatgcgt atatatacca atctaagtct
     1381 gtgctccttc cttcgttctt ccttctgttc ggagattacc gaatcaaaaa aatttcaaag
     1441 aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa ttgaattgaa aagctagctt
     1501 atcgatgata agctgtcaaa gatgagaatt aattccacgg actatagact atactagata
     1561 ctccgtctac tgtacgatac acttccgctc aggtccttgt cctttaacga ggccttacca
     1621 ctcttttgtt actctattga tccagctcag caaaggcagt gtgatctaag attctatctt
     1681 cgcgatgtag taaaactagc tagaccgaga aagagactag aaatgcaaaa ggcacttcta
     1741 caatggctgc catcattatt atccgatgtg acgctgcagc ttctcaatga tattcgaata
     1801 cgctttgagg agatacagcc taatatccga caaactgttt tacagattta cgatcgtact
     1861 tgttacccat cattgaattt tgaacatccg aacctgggag ttttccctga aacagatagt
     1921 atatttgaac ctgtataata atatatagtc tagcgcttta cggaagacaa tgtatgtatt
     1981 tcggttcctg gagaaactat tgcatctatt gcataggtaa tcttgcacgt cgcatccccg
     2041 gttcattttc tgcgtttcca tcttgcactt caatagcata tctttgttaa cgaagcatct
     2101 gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat
     2161 ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta ccaacgaaga
     2221 atctgtgctt catttttgta aaacaaaaat gcaacgcgac gagagcgcta atttttcaaa
     2281 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc
     2341 aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt
     2401 ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt
     2461 gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt
     2521 tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg
     2581 cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc
     2641 gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg
     2701 aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat
     2761 tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac
     2821 tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg
     2881 tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag atacttttga
     2941 gcaatgtttg tggaagcggt attcgcaatg ggaagctcca ccccggttga taatcagaaa
     3001 agccccaaaa acaggaagat tgtataagca aatatttaaa ttgtaattct ttcctgcgtt
     3061 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3121 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagg gtctgacgct
     3181 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3241 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3301 acttggtctg acagtcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa
     3361 tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct
     3421 tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg
     3481 ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca
     3541 tcgtcatggg tcacgacgag atcctcgccg tcgggcatgc tcgccttgag cctggcgaac
     3601 agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg
     3661 gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag
     3721 gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg
     3781 gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag
     3841 tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc
     3901 agccacgata gccgcgctgc ctcgtcttgc agttcattca gggcaccgga caggtcggtc
     3961 ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag
     4021 ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa
     4081 cctgcgtgca atccatcttg ttcaatcata ctcttccttt ttcaatatta ttgaagcatt
     4141 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4201 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg cggacggatc gcttgcctgt
     4261 aacttacacg cgcctcgtgg cgcgcccctg caggatttaa atggccggcc gtttaaacct
     4321 tgtacaccta ccggtcacca gccaacaatg tgcggatggc gttacaactt taaacgtgtg
     4381 gtttatgggt gcaccagggc tttatcgtgt tttatatcga tggcgatttg tgcctccagt
     4441 gtatttttgt atatccaatt aaggtttctt acctaatttt atttttatca tctttagtta
     4501 atgctggttt gctctgtttc tgctgctttc tgtgcggttc tcctcttctc ttgtttcttc
     4561 gtgttgtccc ccatcgccga tgggcttata tggcgtatat atatagagcg agtttttacg
     4621 tcgaagatca tctcagtttg cttgatagcc tttctacttt attactttcg tttttaacct
     4681 cattatactt tagttttctt tgatcggttt ttttctctgt atacttaaaa gttcaaatca
     4741 aagaaacata caaaactacg tttatatcaa ttaatagcgg ccgcatgtct aaaggtgaag
     4801 aattattcac tggtgttgtc ccaattttgg ttgaattaga tggtgatgtt aatggtcaca
     4861 aattttctgt ctccggtgaa ggtgaaggtg atgctactta cggtaaattg accttaaaat
     4921 ttatttgtac tactggtaaa ttgccagttc catggccaac cttagtcact actttcggtt
     4981 atggtgttca atgttttgct agatacccag atcatatgaa acaacatgac tttttcaagt
     5041 ctgccatgcc agaaggttat gttcaagaaa gaactatttt tttcaaagat gacggtaact
     5101 acaagaccag agctgaagtc aagtttgaag gtgatacctt agttaataga atcgaattaa
     5161 aaggtattga ttttaaagaa gatggtaaca ttttaggtca caaattggaa tacaactata
     5221 actctcacaa tgtttacatc atggctgaca aacaaaagaa tggtatcaaa gttaacttca
     5281 aaattagaca caacattgaa gatggttctg ttcaattagc tgaccattat caacaaaata
     5341 ctccaattgg tgatggtcca gtcttgttac cagacaacca ttacttatcc actcaatctg
     5401 ccttatccaa agatccaaac gaaaagagag accacatggt cttgttagaa tttgttactg
     5461 ctgctggtat tacccatggt atggatgaat tgtacaaata agcggccgct cacacttgaa
     5521 tgtgctgcac tttactagaa gtttcttttt ctttttttaa aaataaaaaa agaggagaaa
     5581 aatgctgttt attctgacct tttacataaa tttttcttaa ttgtatctat ctaataacta
     5641 atcaatataa taataataat actaataata ataataataa taataataat agtagtagta
     5701 gtagtagtag taataatgat gatgatgatg atgatggatg atcaaaagcc acaaaaaaga
     5761 atgctccgag aataacctcg gcggttattt cacgtcccgg aggaaaaaaa gtttgataag
     5821 aaagtaaaaa tatagctttt caattcaatt catcattttt tttttattct tttttttgat
     5881 ttcggtttct ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga
     5941 aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt
     6001 gcccagtatt cttaacccaa ctgcacagaa caaaaacctg caggaaacga agataaatca
     6061 tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt tgagatttaa
     6121 atggcgcgcc gtttaaaccc tgcaggggcc ggccccgtag aaaagatcaa aggatcttct
     6181 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6241 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6301 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6361 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6421 gccagtg
//
