LOCUS       sequence-392381-        6761 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresesses mAID-Nb(LaM8) under the control of ADH1 promoter in 
            budding yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6761)
  AUTHORS   Ogawa Y, Nishimura K, Obara K, Kamura T
  TITLE     Development of AlissAID system targeting GFP or mCherry fusion 
            protein.
  JOURNAL   PLoS Genet. 2023 Jun 14;19(6):e1010731. doi: 
            10.1371/journal.pgen.1010731. eCollection 2023 Jun.
  PUBMED    37315088
REFERENCE   2  (bases 1 to 6761)
  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"; journalName: "PLoS 
            Genet."; date: "2023-06-14"; volume: "19(6):e1010731. doi"; pages: "
            10.1371/journal.pgen.1010731. eCollection 2023 Jun"
COMMENT     Sequence Label: pRS305-mAID-LaM8
FEATURES             Location/Qualifiers
     source          1..6761
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        33..51
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     79..95
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     80..96
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        403..804
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     primer_bind     complement(1447..1466)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1448..1466)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1473..1490)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1473..1489)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1482..1504)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1630..2085
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1717..1736)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1927..1948
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2386..2793
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2794..3888
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(4382..4401)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4501..4523
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4561..4579)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4647..4751
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4752..4820,4821..5612)
                     /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     complement(4970..4989)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5783..6371
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6272..6291
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6525..6542
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6659..6680
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6695..6700,6701..6718,6719..6725)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6733..6749
                     /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)."
     primer_bind     6738..6760
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6757..6761,1..12)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6757..6761,1..12)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aacagctatg accatgatta cgccaagctc gaaattaacc ctcactaaag ggaacaaaag
       61 ctggtaccgg gccccccctc gaggtcgacg gtatcgataa gcttgatatc gaattcgggt
      121 gtacaatatg gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata
      181 ccttcgttgg tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac
      241 cagacaagac ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg
      301 tacataacga actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca
      361 ctaccctttt tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct
      421 ttttttttct tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat
      481 gatggaagac actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt
      541 ccagagctga tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca
      601 cactactctc taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata
      661 aaaaaaagtt tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt
      721 ttcctcgtca ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa
      781 gctataccaa gcatacaatc aactatctca tatacaacta gtatgaggga gaggagtgct
      841 tgtcctagag atccagccag acctccggcc agggcacaag ttgtgggatg gccaccggtg
      901 agatcatacc ggcggaacgt gatggtttcc tgccaaagat caagcggtgg cccggaggcg
      961 gcggcgttcg tgagggtatc aatggacgga gcaccgtact tgaggagaat cgatttgagg
     1021 atgtatagag gagctggtgc aggcgctgga gcgggtgccg atgttcagct ggttgaaagc
     1081 ggtggcggct tggtccaggc aggtggaagt ttgcggctta gctgcgctgt gagtgggcga
     1141 cctttttcag agtacaacct gggatggttt cggcaggccc ccggtcggga acgggagttc
     1201 gttgcccgca tcagatccag cgggactacc gtgtataccg actccgtccg gggtagattc
     1261 tctgcaagta gggacaacgc tcggaacatg gggtatctcc agcttaatag tctggagccc
     1321 gaggataccg ccgtatatta ctgtgccatg tccagggtcg atactgattc tcctgccttt
     1381 tatgactatt ggggtcaggg cacccaggtt accgttagca gctaaccgcg gtggagctcc
     1441 aattcgccct atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac
     1501 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     1561 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     1621 ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     1681 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     1741 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     1801 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     1861 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     1921 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     1981 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     2041 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt
     2101 tctccttacg catctgtgcg gtatttcaca ccgcatatcg accggtcgag gagaacttct
     2161 agtatatcta catacctaat attattgcct tattaaaaat ggaatcccaa caattacatc
     2221 aaaatccaca ttctcttcaa aatcaattgt cctgtacttc cttgttcatg tgtgttcaaa
     2281 aacgttatat ttataggata attatactct atttctcaac aagtaattgg ttgtttggcc
     2341 gagcggtcta aggcgcctga ttcaagaaat atcttgaccg cagttaactg tgggaatact
     2401 caggtatcgt aagatgcaag agttcgaatc tcttagcaac cattattttt ttcctcaaca
     2461 taacgagaac acacaggggc gctatcgcac agaatcaaat tcgatgactg gaaatttttt
     2521 gttaatttca gaggtcgcct gacgcatata cctttttcaa ctgaaaaatt gggagaaaaa
     2581 ggaaaggtga gagcgccgga accggctttt catatagaat agagaagcgt tcatgactaa
     2641 atgcttgcat cacaatactt gaagttgaca atattattta aggacctatt gttttttcca
     2701 ataggtggtt agcaatcgtc ttactttcta acttttctta ccttttacat ttcagcaata
     2761 tatatatata tatttcaagg atataccatt ctaatgtctg cccctaagaa gatcgtcgtt
     2821 ttgccaggtg accacgttgg tcaagaaatc acagccgaag ccattaaggt tcttaaagct
     2881 atttctgatg ttcgttccaa tgtcaagttc gatttcgaaa atcatttaat tggtggtgct
     2941 gctatcgatg ctacaggtgt tccacttcca gatgaggcgc tggaagcctc caagaaggct
     3001 gatgccgttt tgttaggtgc tgtgggtggt cctaaatggg gtaccggtag tgttagacct
     3061 gaacaaggtt tactaaaaat ccgtaaagaa cttcaattgt acgccaactt aagaccatgt
     3121 aactttgcat ccgactctct tttagactta tctccaatca agccacaatt tgctaaaggt
     3181 actgacttcg ttgttgtcag agaattagtg ggaggtattt actttggtaa gagaaaggaa
     3241 gacgatggtg atggtgtcgc ttgggatagt gaacaataca ccgttccaga agtgcaaaga
     3301 atcacaagaa tggccgcttt catggcccta caacatgagc caccattgcc tatttggtcc
     3361 ttggataaag ctaatgtttt ggcctcttca agattatgga gaaaaactgt ggaggaaacc
     3421 atcaagaacg aattccctac attgaaggtt caacatcaat tgattgattc tgccgccatg
     3481 atcctagtta agaacccaac ccacctaaat ggtattataa tcaccagcaa catgtttggt
     3541 gatatcatct ccgatgaagc ctccgttatc ccaggttcct tgggtttgtt gccatctgcg
     3601 tccttggcct ctttgccaga caagaacacc gcatttggtt tgtacgaacc atgccacggt
     3661 tctgctccag atttgccaaa gaataaggtc aaccctatcg ccactatctt gtctgctgca
     3721 atgatgttga aattgtcatt gaacttgcct gaagaaggta aggccattga agatgcagtt
     3781 aaaaaggttt tggatgcagg tatcagaact ggtgatttag gtggttccaa cagtaccacc
     3841 gaagtcggtg atgctgtcgc cgaagaagtt aagaaaatcc ttgcttaaaa agattctctt
     3901 tttttatgat atttgtacat aaactttata aatgaaattc ataatagaaa cgacacgaaa
     3961 ttacaaaatg gaatatgttc atagggtaga cgaaactata tacgcaatct acatacattt
     4021 atcaagaagg agaaaaagga ggatgtaaag gaatacaggt aagcaaattg atactaatgg
     4081 ctcaacgtga taaggaaaaa gaattgcact ttaacattaa tattgacaag gaggagggca
     4141 ccacacaaaa agttaggtgt aacagaaaat catgaaacta tgattcctaa tttatatatt
     4201 ggaggatttt ctctaaaaaa aaaaaaatac aacaaataaa aaacactcaa tgacctgacc
     4261 atttgatgga gtttaagtca ataccttctt gaaccatttc ccataatggt gaaagttccc
     4321 tcaagaattt tactctgtca gaaacggcct taacgacgta gtcgatatgg tgcactctca
     4381 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     4441 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4501 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     4561 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4621 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4681 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4741 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4801 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4861 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4921 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4981 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5041 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5101 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5161 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5221 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5281 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5341 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5401 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5461 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5521 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5581 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5641 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5701 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5761 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5821 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5881 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5941 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6001 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6061 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6121 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6181 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6241 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6301 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6361 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6421 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     6481 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6541 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     6601 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     6661 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     6721 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg a
//
