LOCUS       V015258                 8702 bp    DNA     circular SYN 03-MAR-2020
DEFINITION  Exported.
ACCESSION   V015258
VERSION     V015258
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8702)
  AUTHORS   Nemeth T.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-JAN-2020) Department of Microbiology, University of
            Szeged, Kozep fasor 52, Szeged, Csongrad 6726, Hungary
REFERENCE   2  (bases 1 to 8702)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
            (16-JAN-2020) Department of Microbiology, University of Szeged,
            Kozep fasor 52, Szeged, Csongrad 6726, Hungary"
FEATURES             Location/Qualifiers
     source          1..8702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /label="AmpR promoter"
     CDS             129..986
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1160..1748
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    2036..2057
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2072..2102
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2134..2150
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        2171..2189
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      2222..3071
                     /label="Candida albicans TDH3 promoter"
                     /note="Candida albicans TDH3 promoter"
                     /regulatory_class="promoter"
     protein_bind    3078..3202
                     /label="attR1"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3227..3257
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             3311..3967
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     CDS             4312..4614
                     /label="ccdB"
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(4658..4782)
                     /label="attR2"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      4803..5043
                     /label="Saccharomyces cerevisiae URA3 terminator"
                     /note="Saccharomyces cerevisiae URA3 terminator"
                     /regulatory_class="terminator"
     misc_feature    5064..5437
                     /note="recombination target; Candida parapsilosis NEUT5L
                     sequence upstream"
     misc_feature    5438..5443
                     /label="StuI site for plasmid linearization"
                     /note="StuI site for plasmid linearization"
     misc_feature    5444..5827
                     /note="recombination target; Candida parapsilosis NEUT5L
                     sequence downstream"
     CDS             complement(6055..7173)
                     /gene="LEU2"
                     /label="3-isopropylmalate dehydrogenase"
                     /note="3-isopropylmalate dehydrogenase from Candida
                     maltosa. Accession#: P07139"
     regulatory      complement(7174..8048)
                     /label="Candida maltosa LEU2 promoter"
                     /note="Candida maltosa LEU2 promoter"
                     /regulatory_class="promoter"
     promoter        complement(8063..8081)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8088..8104)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      complement(8245..8700)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cgtgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctcaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagctcga aattaaccct cactaaaggg aacaaaagct ggtaccgggc cccccctcga
     2221 gcacacacat acagacaatt gcgatatttt attttagggt caatttggaa cgtcaccata
     2281 cagtattcag tatgattaat ttgacttgca aaatacaaat cgcgtggttc aattccgaaa
     2341 aattgatagt gttccatgaa aaattacaag gagagccgtg aattataaca aatttactca
     2401 cattttggct ggcaatttct ttaaccccct taattttttt catgcagttt attattttac
     2461 gtagaagctt attctctaaa cactcggtta tccgaatata ttttgctttc agagcaattg
     2521 actgatataa taccatcaat tgaataaatc agtatcatca aatgctagtt tgagttcaat
     2581 aaaaggtata atcggtgtta actgaaaggt attgtctaat taattaggtt gcggtttttg
     2641 gatcagctgt ttcaaatcca ggctcgtggt gcacattttt tatttttgtt gtgatgattt
     2701 tttcttttct ttgcttaata gttagttagt aaaatggata atgacgcaaa attgcttctg
     2761 aatggatgat aatgccccgg gtatggcgtg tcccgtacaa atcatattat gcaaaaatag
     2821 ccaaaataga caaaaatagg tggaaattgt ttcaaccaac ccactcaacc actaaaattt
     2881 gtcttgtcaa agccaattcc aacgaaaatt ggattttttt tgacttaata tatattaacc
     2941 cttgaaattc ccttcaattg gaaaattttt ttaatttatt tatttcttct ttcttttttt
     3001 ctttttcttt ttctttcttt tattcatcac aattgaattc aaatcaatta acatcaacaa
     3061 actttacaat cgattcaaca agtttgtaca aaaaagctga acgagaaacg taaaatgata
     3121 taaatatcaa tatattaaat tagattttgc ataaaaaaca gactacataa tactgtaaaa
     3181 cacaacatat ccagtcacta tggcggccgc attaggcacc ccaggcttta cactttatgc
     3241 ttccggctcg tataatgtgt ggattttgag ttaggatccg tcgagatttt caggagctaa
     3301 ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca
     3361 tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt
     3421 tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc
     3481 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc gtatggcaat
     3541 gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga
     3601 gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct
     3661 acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg
     3721 gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga
     3781 tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta
     3841 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     3901 tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg
     3961 cggggcgtaa agatctggat ccggcttact aaaagccaga taacagtatg cgtatttgcg
     4021 cgctgatttt tgcggtataa gaatatatac tgatatgtat acccgaagta tgtcaaaaag
     4081 aggtatgcta tgaagcagcg tattacagtg acagttgaca gcgacagcta tcagttgctc
     4141 aaggcatata tgatgtcaat atctccggtc tggtaagcac aaccatgcag aatgaagccc
     4201 gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga agggatggct gaggtcgccc
     4261 ggtttattga aatgaacggc tcttttgctg acgagaacag gggctggtga aatgcagttt
     4321 aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt acagagtgat
     4381 attattgaca cgcccgggcg acggatggtg atccccctgg ccagtgcacg tctgctgtca
     4441 gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg
     4501 atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt ggctgatctc
     4561 agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg aatataaatg
     4621 tcaggctccc ttatacacag ccagtctgca ggtcgaccat agtgactgga tatgttgtgt
     4681 tttacagtat tatgtagtct gttttttatg caaaatctaa tttaatatat tgatatttat
     4741 atcattttac gtttctcgtt cagctttctt gtacaaagtg gttgctagca tgaataaata
     4801 gcattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat
     4861 cagttattac ccgggaatct cggtcgtaat gatttctata atgacgaaaa aaaaaaattg
     4921 gaaagaaaaa gcttcatggc ctttataaaa aggaactatc caatacctcg ccagaaccaa
     4981 gtaacagtat tttacggggc acaaatcaag aacaataaga caggactgta aagatggacg
     5041 catagcagat cgaattcacg cgtaacctca tctcaaggcg cacgtttttc aataaatttc
     5101 aataaatttg aaattaatta atccgatcct ccttaattac tttgttgttt gtttacgatt
     5161 gtgataattt tagcaaaatc aaaccacagt tttctgttgt ttctttttta tgttcttgac
     5221 ttgtagaata gctccaaggg aaggaagggg gtgttcgggt gacttttatc cccgtgcaaa
     5281 gagagtagtt tgtacgtagt gtacccctat tgttcactgt atcaatccag ttgccacaat
     5341 ttttgtaaac ttcgctttct aaggcgggtt ttctaatttt ctttttcttg attttctttg
     5401 ttgccttttc tctcgctcca cttcggtcca ggtcgctagg cctccgaccc ccactcctca
     5461 ccccaaaaaa aaaagaaaaa agaattacac acaaaattgg aatcccattc agaaatggaa
     5521 caatcggcac caactttaca aaccattgat attcaagttt acgaaatggt taaccaatga
     5581 agtgattgaa tagttttctc gtgcgatgat tgtatcctcc tccatattag ccctctactt
     5641 cttatgtata tagtaaggaa cagcaattaa tgaagttcaa catgcacatt aacacctcac
     5701 ctattaggcg taatctaaaa attggaattg attgatctcc ccatcttctt atctctctct
     5761 ctctatcaca agaacggcat tttttattat cattatttca ttatgacgca atcatgtatt
     5821 tttgtgtact agtccttacc tacccattct agaaagagcc aatcagaatc aagtgaggct
     5881 accattactt caaatagatt tatttccgaa gatccaagta aaccggtgag agaaatttcc
     5941 gatattgaat tatcgcaatt tttagaagaa tcggtacatt tttaaaaaaa agtatttagg
     6001 tggtgttcat tttgtattaa taatattcgt taaattttgt gtatacttta cttaagcttt
     6061 ttcttttaag attttagtaa cagcttcgac aatcgcgtca ccaacttcag tagctgaact
     6121 ggtacctctt aaatctgcag ttctgatacc tttatccaag acttgtttca cggcttcttc
     6181 taaagcttca gcttctttca cacaatccaa agataatctc aacatagaag cagcagataa
     6241 aatagtagca attgggttaa ctttattggc tggtaaatca ggagcagaac cgtgacatgg
     6301 ttcataaaga ccaaaagcgg tgttggtatc tggcaatgaa gccaaagatg cggatggtaa
     6361 taaacccaag gaacctggaa taactgaagc ttcatcggaa atgatatcac caaacatgtt
     6421 agaagtgatg attataccat tcaatttggt tgggttttga attaaaatca tggcggcgga
     6481 gtcaatcaat tgatgttgaa cagacaaagt tgggaactct tcagaaatca ctttatcgac
     6541 agttcttctc cataatctag aagaagctaa aacattagcc ttatccaatg accagattgg
     6601 taatggtggg ttatgttgta aagccatgaa ggcagccata cgggtgattc tggtgacttc
     6661 atcgacggtg tatttttcag tatcccatgc ggtttgttta tcttcacttt cttcttgttc
     6721 ttcacgatca ccgaaataaa taccaccaac taattcacga acaatgatca agtttgtacc
     6781 tttaacgact tctggtctca atggggataa ttccaataag gagtcactgg caaagttaca
     6841 tggtctgatg ttagcataaa ggttcaattc tttacgaatc tttaataaac cttgttctgg
     6901 acgaacggcg ccagtccccc attttggtcc accaacagca cccaataaga cggcatcaga
     6961 ttttttggca ctttcaagtg cgtcatctgg caatgggaca ccagtggcat caatggcagc
     7021 accaccgatt aaatgatgtt tgaaatcgaa atggattttt tggtaaggag tggcagcttc
     7081 aattgcttct aaaactttga tagcttcatt gacaatttcg gtaccgacgt gatcacctgg
     7141 taaaattgta atagttttag ttttaactga cattgtgggt ttgggtagtg aataaaggaa
     7201 ttaaggatga aggaaaaaaa attcttgagt gaaaaatttc atagcctgag tccctcggat
     7261 ttttttcgtt tattatatat ttttatagtg aacaagaaaa aaaaaattga acccggctgc
     7321 ggaacccacg cacgccgtta caggagttag tcattcttca acaatacgaa cgtattattc
     7381 atctggttca gatacatgaa cgaaagtaca atttccgaaa ttactctata cacgataatt
     7441 acacataaca acacaaatta caaaaaaaca aaaaacctta atgtccactc cattcgatag
     7501 agttcaagtc aataccttct tgaaccattt cccataatgg agacatcttt ctcaagaaat
     7561 caactctttc attaattgct tgaataacat agtcaacttc tgcttcagtg gtgaatctac
     7621 caataccgaa tctaatggaa gaatgagcca aggcatcatc ggcacccaaa gcatgtaaaa
     7681 catacgatgg ttccaaagat gcagaggtac atgcagaacc agaactcaat gcaatatctt
     7741 tcaatgccat taacaaagat tccccttcaa tgtatgcaaa ggaaacattc acacaacctg
     7801 ggtattgata tgttgggtta ttacaaccgt taaactgagt agatggaatg gataacaaac
     7861 cgttcttcaa tttagtagat aacctctcaa tatgtttctt gtcaaatgca gattcttgtt
     7921 tcattaatct tgcagcttca ccaaacccag ctactaatgg tggtgccaaa gtacctgatc
     7981 tcaaacctct ttcttgaccc ccaccagtga tgattggatc caagggcgaa ttccagcaca
     8041 ctggcggcga gctccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt
     8101 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     8161 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     8221 ttgcgcagcc tgaatggcga atggaaattg taagcgttaa tattttgtta aaattcgcgt
     8281 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     8341 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     8401 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     8461 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     8521 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     8581 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     8641 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     8701 ca
//
