LOCUS       62056_18210        8426 bp DNA     circular SYN 03-MAR-2020
DEFINITION  Expression vector pNRVL-caSAT1-ADE2RI, complete sequence.
ACCESSION   MT001913
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8426)
  AUTHORS   Nemeth T.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-JAN-2020) Department of Microbiology, University of 
            Szeged, Kozep fasor 52, Szeged, Csongrad 6726, Hungary
REFERENCE   2  (bases 1 to 8426)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (28-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..8426
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(387..473)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    570..575
                     /label=StuI site for cassette release
                     /note="StuI site for cassette release"
     misc_feature    576..1039
                     /note="site of recombination; CpNEUT5LUpup from C.
                     parapsilosis"
     regulatory      1094..1591
                     /label=Candida albicans ACT1 promoter
                     /note="Candida albicans ACT1 promoter"
                     /regulatory_class="promoter"
     gene            1592..2818
                     /gene="caSAT1"
                     /label=caSAT1
     CDS             join(1592..1601,2256..2818)
                     /codon_start=1
                     /transl_table=11
                     /gene="caSAT1"
                     /product="streptothricin acetyltransferase"
                     /label=caSAT1
                     /note="codon-optimized for Candida albicans"
                     /protein_id="QID92300.1"
                     /translation="MDGEEVAALVIDNGSHMKISVIPEQVAETLDAENHFIVREVFDVH
                     LSDQGFELSTRSVSPYRKDYISDDDSDEDSACYGAFIDQELVGKIELNSTWNDLASIEH
                     IVVSHTHRGKGVAHSLIEFAKKWALSRQLLGIRLETQTNNVPACNLYAKCGFTLGGIDL
                     FTYKTRPQVSNETAMYWYWFSGAQDDA"
     regulatory      2822..2951
                     /label=Candida albicans URA3 terminator
                     /note="Candida albicans URA3 terminator"
                     /regulatory_class="terminator"
     regulatory      complement(2985..3484)
                     /label=C. parapsilosis CPAR2_805940 (ADE2) terminator
                     /note="C. parapsilosis CPAR2_805940 (ADE2) terminator"
                     /regulatory_class="terminator"
     gene            complement(3485..5197)
                     /gene="ADE2"
                     /label=ADE2
                     /note="similar to ADE2 from Saccharomyces cerevisiae"
     CDS             complement(3485..5197)
                     /codon_start=1
                     /transl_table=11
                     /gene="ADE2"
                     /product="phosphoribosylaminoimidazole carboxylase-like
                     protein"
                     /label=ADE2
                     /protein_id="QID92301.1"
                     /translation="MDSKTIGILGGGQLGRMIVEAAHRLNIKTVILDAPQSPAKQINAL
                     DEHVDGSFTDYESIAKLAEKVDILTVEIEHVDVDALKMIQLKFPSIEIYPLPDTIKLIQ
                     DKYLQKEHLIQHGIAVTDSQAIAENTTDNLLKIGQAYGYPYMLKSRTLAYDGRGNFVVK
                     DESYVEKALKFLADRPLYAEKWCPFSKELAVMVVRSIEGEVFSYPTVETIHQNNICHLV
                     YAPARINDTLAEKASILAQNAVKSFPGCGIFGVEMFLLESGELLINEIAPRPHNSGHYT
                     IDACVTSQFEAHVRAVTGLPMPRGFTEFSTTITNAIMLNVLGDKDTPNKELETCRRALE
                     TPHSTVYLYGKTTRPERKMGHINIVTSSMDDAENRLAYIMGKSDKLQASPDANGERQQP
                     LVGIIMGSDSDLPVMAVGARILKQFGVPFELTIVSAHRTPHRMSKYAIEAAQRGLKCII
                     AGAGGAAHLPGMVAAMTPLPVIGVPVKGSTLDGVDSLHSIVQMPRGIPVATVAINNSTN
                     AALLAVRILGAVDSKWLNKMTNYMQNMEEEVLKKAEVVEEIGYEDYLTDKLKK"
     CDS             complement(5800..5820)
                     /label=7xHis
                     /note="6xHis affinity tag"
     misc_feature    6204..6654
                     /note="site of recombination; CpNEUT5LDowndown from C. 
                     parapsilosis"
     misc_feature    6655..6660
                     /label=StuI site for cassette release
                     /note="StuI site for cassette release"
     promoter        complement(6668..6686)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6691..6707)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             6820..7626
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      7776..8364
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagtcaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacactagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccta ggccttgaat tggattggga ttagattagg
      601 atttgatggg tttttttttt tctgttaaat ggttatataa tcttcgtttc tgtgagatgt
      661 tttttttttt ctaaattaaa attgtattat gtaatggatt atgtaatttt gcattgcatt
      721 ttttagttat agtagtagta atagtagtac tgcatataca aacacactat tgtagatgag
      781 ggtagtgtac tattgcacaa atttggttta ctcctatttc gcatcattgc agttcatggt
      841 ttgtatgcag taatatatat ttatctctat ttctgtctct cctaagacca gtacaactta
      901 ccactcaatc acacgatata gtctacgagg ctacttcttt atctaccact ggcaatataa
      961 tggtaaagag agcttttccc aagaaccagg tgtggaataa tgaaaagtct atctatggaa
     1021 ggatacatgt tctagatctg cggccgcagc tcggatccac tagtaacggc cgccagtgtg
     1081 ctggaattcc gagcgtcaaa actagagaat aataaagaaa acgatctttt caaaaagaaa
     1141 aaacctttta gttttccttt gttgttgttg tgggtgtgtg ctatttatat tatatagttt
     1201 actcataata ccataaaata ttcggtttga ttaggttatt ttaataagct aatttgtttc
     1261 taatcgtgta atttatgctg tgtatattaa gtagtgtgtg cactgcccaa aaatgtttgt
     1321 tgtttatagt cggttaaaga gaaaaaagaa aaaaagatcc atacacacac gttaattagt
     1381 tgttcaacgt aatacactca tattttgttc ttatttgctt tcggtcgctg ttctcaccaa
     1441 gatttattgc caacgaaaca attttttttt atatattttc agatttttct ttttttcctt
     1501 tcctttcctt ttctaatttt cactcctggt tttctttctt tcttagaaac attatctcga
     1561 tattaatatt aaaaaaatat aatcattcaa aatggacggt ggtatgtttt agtttagctt
     1621 caattctaat tgattgatta atcagttgat tggtttcaat atgacaaatg ggtagggtgg
     1681 gaaaacttca ttttcaattc agatcaaact tttttgttgt cgacataata tttctcgttt
     1741 gggatgttac tgtcacatta ataatacaca cacatcagct tataattttg aaagtaattt
     1801 atcagatatg ttgtgacgat cagtggaaat ggctaacttc aatgtatctg ttcttcccct
     1861 ttttcaaagt tcacgttttt tgattgattg attgatctgt cggcagtggt ttcaaaacca
     1921 ttcggtgagt aatcctatca atcaatgtta cgacaaaagg ctcaatattc aaaattgcaa
     1981 tgttttatgt tttcctacgt gtacttgtgc aaggcaattg attcaacatt gcttttggtg
     2041 tttgacgagt ttctagtttg gacttgtgtt gttatctggg ctatacagat ttcccggctc
     2101 actatgaatt ttttttttcg acgctcagtg cacacaacta taaacaacac aaacacaaac
     2161 acagcaagaa aaaaaaaaaa cgaacattga attgaaacca agccaactga aaaattcctt
     2221 atttaaatga ctgtcatact aacccatttt tatagaagaa gttgctgctt tagttatcga
     2281 taacggttct catatgaaaa tttcggtgat ccctgagcag gtggcggaaa cattggatgc
     2341 tgagaaccat ttcattgttc gtgaagtgtt cgatgtgcac ctatccgacc aaggctttga
     2401 actatctacc agaagtgtga gcccctaccg gaaggattac atctcggatg atgactctga
     2461 tgaagactct gcttgctatg gcgcattcat cgaccaagag cttgtcggga agattgaact
     2521 caactcaaca tggaacgatc tagcctctat cgaacacatt gttgtgtcgc acacgcaccg
     2581 aggcaaagga gtcgcgcaca gtctcatcga atttgcgaaa aagtgggcac taagcagaca
     2641 gctccttggc atacgattag agacacaaac gaacaatgta cctgcctgca atttgtacgc
     2701 aaaatgtggc tttactctcg gcggcattga cctcttcacg tataaaacta gacctcaagt
     2761 ctcgaacgaa acagcgatgt actggtactg gttctcggga gcacaggatg acgcctaaca
     2821 tatgtgaagt gtgaaggggg agattttcac tttattagat ttgtatatat gtataataaa
     2881 taaataaata agttaaataa ataattagat aagggtggta attattacta tttacaatca
     2941 aaggtggtcc tggaattctg cagatatcca tcacactggc gcgtgtgaaa agaaagggtg
     3001 aatttgctaa atatgccgac gaaagaagta aaagaatagc ggaaagattg gctaaagaaa
     3061 cagctaaaca agaagctgaa aagacaaagg gtaagcaaga taatggccaa gaatcgatgg
     3121 acgtggataa gggaggcgaa gacggtgagg aaggaaagag ccagaaggta agcacttccg
     3181 gatggaggga ctctaattct cagaggtata aacaaagaca actcaagaat aaatccaaaa
     3241 agaagacatt gaagttttaa gagagggtga tcacgtacag tggaatcgct aaatcccacg
     3301 tattgcaaac ccaagcacta ttgattcaat ggaataacca tttgagacaa ttgagaacat
     3361 gcatcagttc agatccacgc tggctcggga tctggattat attcaatgaa tgacatagag
     3421 acggcaaaac taacaaagtt gtatataaat ttaatgtata ataactataa ctttagcaca
     3481 atttttactt tttcaacttg tcagtgaggt aatcttcata cccaatctct tcaacaactt
     3541 cagctttctt caacacttct tcctccatgt tctgcatata attggtcatt ttgttcaacc
     3601 atttcgaatc aactgccccc aagattctta cagctaataa cgcagcattg gtactgttgt
     3661 tgatagctac tgtggcaaca ggaatacccc ttggcatttg cacaatagaa tgcaatgaat
     3721 caacaccgtc taaagtggaa ccttttacgg gcacaccaat cactggtagc ggagtcattg
     3781 cggcaaccat acctggtaaa tgagcagctc caccagcacc agcaatgata catttcaatc
     3841 cacgttgtgc cgcttcaatt gcgtattttg acattctatg tggcgtacga tgagcactca
     3901 caattgttaa ttcaaatgga acaccaaact gtttcaaaat acgagcacca accgccatga
     3961 ctggtaaatc ggaatcagaa cccataatga ttccaactag tggttgttgt ctttcgccat
     4021 tggcatctgg tgaagcttgt agtttatcac ttttacccat tatgtaagcc aatctatttt
     4081 cggcatcatc catcgacgaa gtgacaatgt ttatatggcc cattttacgt tcaggtctgg
     4141 tggttttacc atacaagtat acggttgagt gtggggtttc taatgcgcga cgacacgttt
     4201 ccaattcttt gtttggtgta tctttatcac ccaagacatt caacataatg gcattggtga
     4261 ttgtcgttga aaattcagtg aaaccccttg gcatgggtaa tccagtcact gctctaacat
     4321 gagcttcaaa ctgtgacgtg acacaagcat caattgtata gtgacccgaa ttgtgaggtc
     4381 tcggtgcgat ttcattaatt aaaagctcac cactttctaa taaaaacatc tctactccaa
     4441 atataccaca accagggaaa gatttaacag cattttgagc caaaatcgaa gccttttctg
     4501 ccaaagtatc attgattcta gctggtgcat aaacaagatg acaaatatta ttttggtgaa
     4561 tggtttccac ggtgggatat gaaaacacct caccttcgat tgatctaacc accataactg
     4621 ccaattcttt ggaaaatggg caccattttt cagcatataa cggacgatca gctaaaaact
     4681 ttaaagcttt ctcaacatat gattcatctt tgacaacgaa attacctctt ccgtcatatg
     4741 ccaatgttct cgattttaac atgtatggat atccatacgc ttgaccaatt tttaacaaat
     4801 tgtcagtagt attttcagca atagcttgag aatcagttac agcaatacca tgttgaatca
     4861 aatgttcttt ttgtaaatat ttatcctgta tcaatttaat agtatcaggt aacgggtaaa
     4921 tctcaatact ggggaacttc agttgaatca ttttcaaagc atcaacatca acatgctcaa
     4981 tctcaacagt taaaatgtca accttttcag ccaatttggc aatagattca taatcagtga
     5041 aggatccatc aacatgttca tcaagggcat tgatttgctt ggctggtgat tgcggagcat
     5101 ctaagatgac agttttgata ttaagtctat gagccgcttc cactatcata cgaccaagct
     5161 ggccgcctcc tagtatacct atagttttac tgtccatggt gatgcgaagt aaatggaact
     5221 gagagactgt ttcagggtgg tggtcttttt tctgattaga tatccctatg attacatcag
     5281 agtcacaaaa tttgaaaatt taatttgaaa cggttttttt ttttaactct tcccatccac
     5341 gtcgcacgaa aaaaagtcag aagatgcaaa aagagcatgc ccagaactcc cttttacaga
     5401 acagactttc gatacaagct ttaataatta caaatatact ctgaacaatg aatgataacg
     5461 aaacgcaaaa acgaaaatgt ctagcaaggt gattctctta gagctcaaac atctttaaaa
     5521 gtttctagct ttgaatgaat taagaatgtg aaaggaaaag aagcgtctaa taatcactag
     5581 cataaccttg gatatagccc tgattggtgt atgaatattc ttgaaaactt gctactggct
     5641 ccaagtgtac tggaccagaa ggaaccatca taaggtccat atttgccgat tgatgtagtt
     5701 tgtagcctgg atgatgaggg tgatgataat gggcgggcac ttgttgttgc atttgaatct
     5761 cataatccgg atgatgaaaa tgatgattca ctggtagagg tggtggtggt ggtggtggtg
     5821 gaagggggtt ttgcaatgga ccagcgtcca aaccttgttg tagctgttgt tgtggttgtt
     5881 gttgtggttg ttgtgaatgt aaatcgttgc cagctataaa gtgcggcggg tatcgaagtt
     5941 ctggcacata ttctggattc gatataggaa gcttattggg aaagggcctt gtggtagagg
     6001 cagtggtggc agtagtggca gtggcattgg tagttgttgg cacagttgtc gctgtgttga
     6061 aatcttcaaa acgttcacga taaggtgtcg gcacagaagt agtggcagtg atcctcccag
     6121 gtctaaggtt aggatgcatt gatgatccca cttgtaccgg attagcttcc gcatcataat
     6181 caccagctcc tgtctccctc gagcgaagta gtcctcactg aggagatccc aggctagaaa
     6241 tcagcaaatt ttacatccct catattctga cacgagttta actacatagc aattgtaaaa
     6301 gggctcttgt tttttttttt tttttgtggc ggtgcagtac gtacacgatc tttaaagatt
     6361 agggctataa ccatgtcaaa gtgactcatt ttttttgttc aaatgcgcac tcggatttaa
     6421 agcagcaaaa aaatacttcg ttacaaaaat ggaagcgcat tttccgaaaa aggttaattc
     6481 ctagcgattt tattattttt gttttattta ttaatttttc ttgttttgct attgcatttt
     6541 tgcaaccaaa aatagaaaat tctcgttttt ttttgttgta gcgcaaccaa tttctggtta
     6601 ttcttttttt tttgcaattt ttaaatttgc acgtgacagt gttagtgtta gtgcaggcct
     6661 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
     6721 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     6781 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     6841 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     6901 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     6961 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     7021 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     7081 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     7141 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     7201 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     7261 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     7321 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     7381 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     7441 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     7501 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     7561 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     7621 tttttctaat cagaattggt taattggttg taacactggc agagcattac gctgacttga
     7681 cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     7741 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7801 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7861 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7921 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7981 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     8041 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     8101 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     8161 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     8221 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     8281 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     8341 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     8401 ttttgctggc cttttgctca catgtt
//
