LOCUS       Exported                7177 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  SUC2 regulated expression of an antisense RNAi construct for GFP.
ACCESSION   .
VERSION     .
KEYWORDS    PSUC2-GFPantisense-CYC1t-pRS413
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7177)
  AUTHORS   Williams TC, Espinosa MI, Nielsen LK, Vickers CE
  TITLE     Dynamic regulation of gene expression using sucrose responsive 
            promoters and RNA interference in Saccharomyces cerevisiae.
  JOURNAL   Microb Cell Fact. 2015 Apr 1;14:43. doi: 10.1186/s12934-015-0223-7.
  PUBMED    25886317
REFERENCE   2  (bases 1 to 7177)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7177
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        317..503
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             504..1163
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(1423..1878)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1560..1581)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1772..1791
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2008..2030
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2022..2039
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2023..2039
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2049..2068
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2049..2067
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(3349..4059)
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(4067..4083)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4067..4083)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      4107..4356
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4123..4141)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4403..4423)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4403..4421)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4442..4458)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4442..4458)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4455..4477)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4466..4482
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     promoter        complement(4490..4520)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4673..4690)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4844..5432)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4924..4943)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5603..6463)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6226..6245
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6464..6568)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6605..7108
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     7150..7168
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgcgtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaatt cccgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt
      241 gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta
      301 ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat
      361 tttttttttt cccctagcgg atgactcttt ttttttctta gcgattggca ttatcacata
      421 atgaattata cattatataa agtaatgtga tttcttcgaa gaatatacta aaaaatgagc
      481 aggcaagata aacgaaggca aagatgacag agcagaaagc cctagtaaag cgtattacaa
      541 atgaaaccaa gattcagatt gcgatctctt taaagggtgg tcccctagcg atagagcact
      601 cgatcttccc agaaaaagag gcagaagcag tagcagaaca ggccacacaa tcgcaagtga
      661 ttaacgtcca cacaggtata gggtttctgg accatatgat acatgctctg gccaagcatt
      721 ccggctggtc gctaatcgtt gagtgcattg gtgacttaca catagacgac catcacacca
      781 ctgaagactg cgggattgct ctcggtcaag cttttaaaga ggccctactg gcgcgtggag
      841 taaaaaggtt tggatcagga tttgcgcctt tggatgaggc actttccaga gcggtggtag
      901 atctttcgaa caggccgtac gcagttgtcg aacttggttt gcaaagggag aaagtaggag
      961 atctctcttg cgagatgatc ccgcattttc ttgaaagctt tgcagaggct agcagaatta
     1021 ccctccacgt tgattgtctg cgaggcaaga atgatcatca ccgtagtgag agtgcgttca
     1081 aggctcttgc ggttgccata agagaagcca cctcgcccaa tggtaccaac gatgttccct
     1141 ccaccaaagg tgttcttatg tagtgacacc gattatttaa agctgcagca tacgatatat
     1201 atacatgtgt atatatgtat acctatgaat gtcagtaagt atgtatacga acagtatgat
     1261 actgaagatg acaaggtaat gcatcattct atacgtgtca ttctgaacga ggcgcgcttt
     1321 ccttttttct ttttgctttt tctttttttt tctcttgaac tcgacggatc tatgcggtgt
     1381 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta aacgttaata
     1441 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     1501 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     1561 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     1621 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     1681 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     1741 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     1801 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     1861 cgccgctaca gggcgcgtcg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     1921 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     1981 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     2041 agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccaca tactaagaca
     2101 tttaccgtat gggagttgtt gtcctagcgt agttctcgct cccccagcaa agctcaaaaa
     2161 agtacgtcat ttagaatagt ttgtgagcaa attaccagtc ggtatgctac gttagaaagg
     2221 cccacagtat tcttctacca aaggcgtgcc tttgttgaac tcgatccatt atgagggctt
     2281 ccattattcc ccgcattttt attactctga acaggaataa aaagaaaaaa cccagtttag
     2341 gaaattatcc gggggcgaag aaatacgcgt agcgttaatc gaccccacgt ccagggtttt
     2401 tccatggagg tttctggaaa aactgacgag gaatgtgatt ataaatccct ttatgtgatg
     2461 tctaagactt ttaaggtacg cccgatgttt gcctattacc atcatagaga cgtttctttt
     2521 cgaggaatgc ttaaacgact ttgtttgaca aaaatgttgc ctaagggctc tatagtaaac
     2581 catttggaag aaagatttga cgactttttt tttttggatt tcgatcctat aatccttcct
     2641 cctgaaaaga aacatataaa tagatatgta ttattcttca aaacattctc ttgttcttgt
     2701 gctttttttt taccatatat cttacttttt tttttctctc agagaaacaa gcaaaacaaa
     2761 aagcttttct tttcactaac gtatatgatc gataagcttg atatcgaatt cctatattac
     2821 ttgggtattg cccataccaa aagaaattaa agatgaggca ctgctagatt taccgctatt
     2881 tatggtccca gttggcgagg ggaacattcc atggttaatg aagcttgtct tgggtgggcc
     2941 cccattgagg taatgcgccg ttgcgggatt ttggttttcc ttgttagact catttaggtt
     3001 tagcaaatca atataatcat tggttgcgtt attgctgtta ggacccgtga tttgttcgta
     3061 gatcctttgt ttgttatttg tattcaccag actattcaca ctaacggtgc taccacatat
     3121 actgtttgac tgctgctgac caaattggta catacttgga gtgattggtg atgattgttc
     3181 catattccgg ctgaaaacgc tgttatcaga gagtgagctc atgtttctca ttggagtgtg
     3241 gctttgagat gaggaagcgg gcgaaggaat ggatgtgcta tttgaatttt cgaaagagca
     3301 tgatggggtt gatatggtaa gctttctcga aatgttcaag ttggatgctt tgtacaattc
     3361 atccatacca tgggtaatac cagcagcagt aacaaattct aacaagacca tgtggtctct
     3421 cttttcgttt ggatctttgg ataaggcaga ttgagtggat aagtaatggt tgtctggtaa
     3481 caagactgga ccatcaccaa ttggagtatt ttgttgataa tggtcagcta attgaacaga
     3541 accatcttca atgttgtgtc taattttgaa gttaactttg ataccattct tttgtttgtc
     3601 agccatgatg taaacattgt gagagttata gttgtattcc aatttgtgac ctaaaatgtt
     3661 accatcttct ttaaaatcaa taccttttaa ttcgattcta ttaactaagg tatcaccttc
     3721 aaacttgact tcagctctgg tcttgtagtt accgtcatct ttgaaaaaaa tagttctttc
     3781 ttgaacataa ccttctggca tggcagactt gaaaaagtca tgttgtttca tatgatctgg
     3841 gtatctcgca aaacattgaa caccataacc gaaagtagtg actaaggttg gccatggaac
     3901 tggcaattta ccagtagtac aaataaattt taaggtcaat ttaccgtaag tagcatcacc
     3961 ttcaccttca ccggagacag aaaatttgtg accattaaca tcaccatcta attcaaccaa
     4021 aattgggaca acaccagtga ataattcttc acctttagac ccgggggatc cactagttct
     4081 agaaggcccc ttttcctttg tcgatatcat gtaattagtt atgtcacgct tacattcacg
     4141 ccctcctccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt
     4201 ccctatttat tttttttaat agttatgtta gtattaagaa cgttatttat atttcaaatt
     4261 tttctttttt ttctgtacaa acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt
     4321 gagaaggttt tgggacgctc gaaggcttta atttgcaagc ttcgcagttt acactctcat
     4381 cgtcgagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     4441 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacataggag
     4501 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gaggtaactc acattaattg
     4561 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4621 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4681 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4741 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4801 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4861 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4921 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4981 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5041 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5101 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5161 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5221 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5281 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5341 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5401 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5461 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5521 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5581 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5641 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5701 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5761 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5821 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5881 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5941 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6001 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6061 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6121 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6181 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6241 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6301 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6361 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6421 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6481 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6541 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggtcct
     6601 tttcatcacg tgctataaaa ataattataa tttaaatttt ttaatataaa tatataaatt
     6661 aaaaatagaa agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc cgaagatgta
     6721 aaagactcta gggggatcgc caacaaatac taccttttat cttgctcttc ctgctctcag
     6781 gtattaatgc cgaattgttt catcttgtct gtgtagaaga ccacacacga aaatcctgtg
     6841 attttacatt ttacttatcg ttaatcgaat gtatatctat ttaatctgct tttcttgtct
     6901 aataaatata tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt gtttattttt
     6961 ttttcttcat tccgtaactc ttctaccttc tttatttact ttctaaaatc caaatacaaa
     7021 acataaaaat aaataaacac agagtaaatt cccaaattat tccatcatta aaagatacga
     7081 ggcgcgtgta agttacaggc aagcgatccg tcctaagaaa ccattattat catgacatta
     7141 acctataaaa ataggcgtat cacgaggccc tttcgtc
//
