LOCUS       62056_8340        7691 bp DNA     circular SYN 03-MAR-2020
DEFINITION  Cloning vector pDCpOE-N-N5L, complete sequence.
ACCESSION   MN974376
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7691)
  AUTHORS   Nemeth T.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-JAN-2020) Department of Microbiology, University of 
            Szeged, Kozep fasor 52, Szeged, Csongrad 6726, Hungary
REFERENCE   2  (bases 1 to 7691)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (20-JAN-2020) Department of Microbiology, University of Szeged, 
            Kozep fasor 52, Szeged, Csongrad 6726, Hungary"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: CLC Genomics Workbench v. v11
            Sequencing Technology :: IonTorrent
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7691
                     /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
                     /label=Candida parapsilosis NEUT5L sequence upstream
                     /note="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
                     /label=Candida parapsilosis NEUT5L sequence downstream
                     /note="Candida parapsilosis NEUT5L sequence downstream"
     terminator      complement(5854..6051)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(6069..6632)
                     /label=NrsR
                     /note="nourseothricin acetyltransferase"
     promoter        complement(6652..6995)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        complement(7052..7070)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7077..7093)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(7234..7689)
                     /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 atcgctgaga
      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 agtcgttagt atcgaatcga cagcagtata gcgaccagca ttcacatacg
     5881 attgacgcat gatattactt tctgcgcact taacttcgca tctgggcaga tgatgtcgag
     5941 gcgaaaaaaa atataaatca cgctaacatt tgattaaaat agaacaacta caatataaaa
     6001 aaactataca aatgacaagt tcttgaaaac aagaatcttt ttattgtcag tactgactcg
     6061 agttattatg gacatggcat agacatatac aaagcttgtt caccatcgga agcagtacca
     6121 tcgtataaag cagtatccaa accacacaaa gtgaaaccca ttcttctata agcatgaata
     6181 gctggagcat taacattggt aacttccaac cacaaatgac cagcacctct ttctctggcg
     6241 aattcagtag ccaaacccat caaagctcta ccaacaccat gacctctatg ttctggagca
     6301 acttcaatat cttcaacagt caatcttctg ttccaaccag aataagaaac aacaacgaaa
     6361 ccagccaaat caccatcatc accataagca acgaaagttc tagaatctgg atcaccatct
     6421 tcaccagcat cggattcatc atcggattca tcatctggga aaaccttagt caatggtgga
     6481 tcaactggaa cttctctcaa agtgaaacca tcaccagtag cagtaactct aaaaacagta
     6541 tcggtagtga aagaaccatc caaagcttca atagcttcag catcacctgg aacagaagtt
     6601 ctgtatctat aagcagtatc atccaaagta gtagacataa ttgtaggatc cggttgttta
     6661 tgttcggatg tgatgtgaga actgtatcct agcaagattt taaaaggaag tatatgaaag
     6721 aagaacctca gtggcaaatc ctaacctttt atatttctct acaggggcgc ggcgtgggga
     6781 caattcaacg cgtctgtgag gggagcgttt ccctgctcgc aggtctgcag cgaggagccg
     6841 taatttttgc ttcgcgccgt gcggccatca aaatgtatgg atgcaaatga ttatacatgg
     6901 ggatgtatgg gctaaatgta cgggcgacag tcacatcatg cccctgagct gcgcacgtca
     6961 agactgtcaa ggagggtatt ctgggcctcc atgtcgctgg ccgggtgacc cggcggggac
     7021 gaggcaagct tgatatcgag ctccaattcg ccctatagtg agtcgtatta caattcactg
     7081 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     7141 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     7201 tcccaacagt tgcgcagcct gaatggcgaa tggaaattgt aagcgttaat attttgttaa
     7261 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     7321 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     7381 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     7441 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     7501 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     7561 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     7621 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     7681 agggcgcgtc a
//
