LOCUS       sequence-6841-ex        6558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6558)
  TITLE     Unpublished, Paco
REFERENCE   2  (bases 1 to 6558)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pYES2.1-TRP1
FEATURES             Location/Qualifiers
     source          1..6558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..443
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2..119
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     414..437
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     475..494
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        475..493
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             513..1184
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     CDS             1215..1256
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1266..1283
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1304..1551
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1320..1338)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1628..1645)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1799..2387)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1879..1898)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2558..3349,3350..3418))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLID*MEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             complement(3514..4317)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4318..4538)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      5137..6017
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(6086..6541)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     6435..6454
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 acggattaga agccgccgag cgggtgacag ccctccgaag gaagactctc ctccgtgcgt
       61 cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga
      121 acaataaaga ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac
      181 ctggccccac aaaccttcaa atgaacgaat caaattaaca accataggat gataatgcga
      241 ttagtttttt agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat
      301 taacagatat ataaatgcaa aaactgcata accactttaa ctaatacttt caacattttc
      361 ggtttgtatt acttcttatt caaatgtaat aaaagtatca acaaaaaatt gttaatatac
      421 ctctatactt taacgtcaag gagaaaaaac cccggatcgg actactagca gctgtaatac
      481 gactcactat agggaatatt aagctcgccc ttatgtctgt tattaatttc acaggtagtt
      541 ctggtccatt ggtgaaagtt tgcggcttgc agagcacaga ggccgcagaa tgtgctctag
      601 attccgatgc tgacttgctg ggtattatat gtgtgcccaa tagaaagaga acaattgacc
      661 cggttattgc aaggaaaatt tcaagtcttg taaaagcata taaaaatagt tcaggcactc
      721 cgaaatactt ggttggcgtg tttcgtaatc aacctaagga ggatgttttg gctctggtca
      781 atgattacgg cattgatatc gtccaactgc atggagatga gtcgtggcaa gaataccaag
      841 agttcctcgg tttgccagtt attaaaagac tcgtatttcc aaaagactgc aacatactac
      901 tcagtgcagc ttcacagaaa cctcattcgt ttattccctt gtttgattca gaagcaggtg
      961 ggacaggtga acttttggat tggaactcga tttctgactg ggttggaagg caagagagcc
     1021 ccgaaagctt acattttatg ttagctggtg gactgacgcc agaaaatgtt ggtgatgcgc
     1081 ttagattaaa tggcgttatt ggtgttgatg taagcggagg tgtggagaca aatggtgtaa
     1141 aagactctaa caaaatagca aatttcgtca aaaatgctaa gaaaaagggc gagcttcgag
     1201 gtcacccatt cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta
     1261 ccggtcatca tcaccatcac cattgagttt ctagagggcc gcatcatgta attagttatg
     1321 tcacgcttac attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga
     1381 caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta
     1441 tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata
     1501 ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg caagctgcgg
     1561 ccctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     1621 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1681 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1741 tgtgagcaaa aggccagcaa aagcccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1801 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1861 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1921 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1981 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2041 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2101 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2161 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2221 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2281 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2341 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2401 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2461 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2521 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2581 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2641 agataactac gatacgggag cgcttaccat ctggccccag tgctgcaatg ataccgcgag
     2701 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2761 gcagaagtgg tcctgcaact ttatccgcct ccattcagtc tattaattgt tgccgggaag
     2821 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttggcatt gctacaggca
     2881 tcgtggtgtc actctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2941 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3001 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3061 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3121 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3181 ataatagtgt atcacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3241 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3301 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3361 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3421 tcttcctttt tcaatgggta ataactgata taattaaatt gaagctctaa tttgtgagtt
     3481 tagtatacat gcatttactt ataatacagt tttttagttt tgctggccgc atcttctcaa
     3541 atatgcttcc cagcctgctt ttctgtaacg ttcaccctct accttagcat cccttccctt
     3601 tgcaaatagt cctcttccaa caataataat gtcagatcct gtagagacca catcatccac
     3661 ggttctatac tgttgaccca atgcgtctcc cttgtcatct aaacccacac cgggtgtcat
     3721 aatcaaccaa tcgtaacctt catctcttcc acccatgtct ctttgagcaa taaagccgat
     3781 aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga
     3841 tagggagccc ttgcatgaca attctgctaa catcaaaagg cctctaggtt cctttgttac
     3901 ttcttctgcc gcctgcttca aaccgctaac aatacctggg cccaccacac cgtgtgcatt
     3961 cgtaatgtct gcccattctg ctattctgta tacacccgca gagtactgca atttgactgt
     4021 attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact tggcggataa
     4081 tgcctttagc ggcttaactg tgccctccat ggaaaaatca gtcaagatat ccacatgtgt
     4141 ttttagtaaa caaattttgg gacctaatgc ttcaactaac tccagtaatt ccttggtggt
     4201 acgaacatcc aatgaagcac acaagtttgt ttgcttttcg tgcatgatat taaatagctt
     4261 ggcagcaaca ggactaggat gagtagcagc acgttcctta tatgtagctt tcgacatgat
     4321 ttatcttcgt ttcctgcagg tttttgttct gtgcagttgg gttaagaata ctgggcaatt
     4381 tcatgtttct tcaacactac atatgcgtat atataccaat ctaagtctgt gctccttcct
     4441 tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa tttcaaagaa accgaaatca
     4501 aaaaaaagaa taaaaaaaaa atgatgaatt gaattgaaaa gctagcttat cgatgataag
     4561 ctgtcaaaga tgagaattaa ttccacggac tatagactat actagatact ccgtctactg
     4621 tacgatacac ttccgctcag gtccttgtcc tttaacgagg ccttaccact cttttgttac
     4681 tctattgatc cagctcagca aaggcagtgt gatctaagat tctatcttcg cgatgtagta
     4741 aaactagcta gaccgagaaa gagactagaa atgcaaaagg cacttctaca atggctgcca
     4801 tcattattat ccgatgtgac gctgcagctt ctcaatgata ttcgaatacg ctttgaggag
     4861 atacagccta atatccgaca aactgtttta cagatttacg atcgtacttg ttacccatca
     4921 ttgaattttg aacatccgaa cctgggagtt ttccctgaaa cagatagtat atttgaacct
     4981 gtataataat atatagtcta gcgctttacg gaagacaatg tatgtatttc ggttcctgga
     5041 gaaactattg catctattgc ataggtaatc ttgcacgtcg catccccggt tcattttctg
     5101 cgtttccatc ttgcacttca atagcatatc tttgttaacg aagcatctgt gcttcatttt
     5161 gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     5221 tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca
     5281 tttttgtaaa acaaaaatgc aacgcgacga gagcgctaat ttttcaaaca aagaatctga
     5341 gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct
     5401 atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat
     5461 cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg
     5521 cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa
     5581 aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt
     5641 ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg
     5701 aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc
     5761 tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc
     5821 actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca
     5881 taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg
     5941 ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg
     6001 gaagcggtat tcgcaatggg aagctccacc ccggttgata atcagaaaag ccccaaaaac
     6061 aggaagattg tataagcaaa tatttaaatt gtaaacgtta atattttgtt aaaattcgcg
     6121 ttaaattttt gttaaatcag ctcatttttt aacgaatagc ccgaaatcgg caaaatccct
     6181 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttc caacaagagt
     6241 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaagggtcta tcagggcgat
     6301 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     6361 gtaaatcgga agggtaaacg gatgccccca tttagagctt gacggggaaa gccggcgaac
     6421 gtggcgagaa aggaagggaa gaaagcgaaa ggagcggggg ctagggcggt gggaagtgta
     6481 ggggtcacgc tgggcgtaac caccacaccc gccgcgctta atggggcgct acagggcgcg
     6541 tggggatgat ccactagt
//
