LOCUS       sequence-191147-        8736 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMYA5 (aa 3859 – 4069) Leu allele (4063) in a GAL4 AD vector, used 
            for yeast-two hybrid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8736)
  TITLE     Shiang Lab CMYA5 project plasmids (unpublished)
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8736)
  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: 
            "Unpublished"
COMMENT     Sequence Label: CMYA5tnT/pACT2
FEATURES             Location/Qualifiers
     source          1..8736
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    112..145
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    223..256
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    300..333
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    411..444
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      complement(465..652)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     573..597
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(1699..1725)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             complement(1705..1731)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1744..2085)
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     CDS             complement(2101..2121)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(2167..2568)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     primer_bind     complement(2597..2619)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2705..2845
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2760..2779
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2860..2877)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3031..3619)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3111..3130)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3790..4581,4582..4650))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4413..4432
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4651..4755)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5020..6362
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    5352..5399
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        6839..7244
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             7245..8339
                     /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"
ORIGIN
        1 ttcttgaacc atttcccata atggtgaaag ttccctcaag aattttactc tgtcagaaac
       61 ggccttaacg acgtagtcga ccaattccga tcatattcaa taacccttaa tataacttcg
      121 tataatgtat gctatacgaa gttattaggt ctgaagagga gtttacgtcc agccaagcta
      181 gcttggctgc aggtcgagcg gccgcgatcc ggaaccctta atataacttc gtataatgta
      241 tgctatacga agttattagg tccctcgacc aattccgatc atattcaata acccttaata
      301 taacttcgta taatgtatgc tatacgaagt tattaggtct gaagaggagt ttacgtccag
      361 ccaagctagc ttggctgcag gtcgagcggc cgcgatccgg aacccttaat ataacttcgt
      421 ataatgtatg ctatacgaag ttattaggtc cctcgaccgg catgccggta gaggtgtggt
      481 caataagagc gacctcatgc tatacctgag aaagcaacct gacctacagg aaagagttac
      541 tcaagaataa gaattttcgt tttaaaacct aagagtcact ttaaaatttg tatacactta
      601 ttttttttat aacttattta ataataaaaa tcataaatca taagaaattc gcttatttag
      661 aagtgtcaac aacgtatcta ccaacgattt gacccttttc catcttttcg taaatttctg
      721 gcaaggtaga caagccgaca accttgattg gagacttgac caaacctctg gcgaagaagt
      781 ccaaagcttc tgaataagcc ctcgtaatat attttcatga agaatttagg tccaaaaaaa
      841 agatgggcat taattctagt catttaaaaa attctataga tcagaggtta catggccaag
      901 attgaaactt agaggagtat agttacataa aagaaggcaa aacgatgtat aaatgaaaga
      961 aattgagatg gtgcacgatg cacagttgaa gtgaacttgc ggggtttttc agtatctacg
     1021 attcatagat ctctcgagct cgaattcctt gtgccttaca gaatccaggg gctctatccc
     1081 caggtgcaaa gtacattttc caggtttctc cagggcaaag gcagggtgga caccctcatt
     1141 aaacctgtgc ctgatgatga agagcaactg ctcgctctct gcgttgatga agataagtct
     1201 ctggttgttg tagtccagca ggatgcccac tctggctgga cgctcagtca catgaacatc
     1261 actcacaata ccactgtaga aaaatgtgta tctctgtggc tcagagcagt gcatgtacca
     1321 tgaggtctcc ccttgtccta gggcacctgc ctgcacagcc gagctggagc agatgccgag
     1381 tcgatatgct gggcagtctg tgactgtggt ttcccagtaa tgctggccac aggaaggcag
     1441 ctcctcaccc agcactgacg ggatttccgt cagccgtctc ctctcaccaa agctgatcac
     1501 tgtcccactt gaggaaatgt gtagagcagg atgagctgtt tctctcaaca agagaaatct
     1561 ggttcctctg gtggagatga gagcagcttc actctgttca cttgtcccaa atgcattgat
     1621 ggccctcaca tagaaaaagt agttatcatt gggttggagg ttaactttca gctggagtcc
     1681 catggccata tgaccaccca agctagcgta atctggaaca tcgtatgggt aagccataga
     1741 gatctctttt tttgggtttg gtggggtatc ttcatcatcg aatagatagt tatatacatc
     1801 atccattgta gtggtattaa acatccctgt agtgattcca aacgcgttat acgcagtttg
     1861 gtccgtccaa ccaggtgaca gtggttttga attattacca tcatcaattt tactagccgt
     1921 gatttcatta ttcatgaagt tatcatgaac gttagaggag gcaattggtt gtgaaagcgc
     1981 ttgagaattt gtttgagttg ttatgaggtt cggaccgttg ctactgttag tgaaagtgaa
     2041 ggacaatgag ctatcagcaa tattcccact ttgattaaaa ttggcggcgg tacccaattc
     2101 gacctttctc ttcttttttg gaggctcggg aattaattcc gctttatcca tctttgcaaa
     2161 gcttggagtt gattgtatgc ttggtatagc ttgaaatatt gtgcagaaaa agaaacaagg
     2221 aagaaaggga acgagaacaa tgacgaggaa acaaaagatt aataattgca ggtctattta
     2281 tacttgatag caagacagca aacttttttt tatttcaaat tcaagtaact ggaaggaagg
     2341 ccgtataccg ttgctcatta gagagtagtg tgcgtgaatg aaggaaggaa aaagtttcgt
     2401 gtgcttcgag atacccctca tcagctctgg aacaacgaca tctgttggtg ctgtctttgt
     2461 cgttaatttt ttcctttagt gtcttccatc atttttttgt cattgcggat atggtgagac
     2521 aacaacgggg gagagagaaa agaaaaaaaa agaaaagaag ttgctgcctc gcgcgtttcg
     2581 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     2641 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     2701 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc
     2761 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     2821 cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2881 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2941 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3001 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3061 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3121 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3181 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3241 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3301 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3361 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3421 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     3481 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3541 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3601 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3661 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3721 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3781 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3841 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3901 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3961 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4021 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4081 tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat
     4141 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4201 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4261 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4321 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4381 accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt
     4441 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4501 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4561 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4621 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4681 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4741 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gagctttgaa gaaaaatgcg
     4801 ccttattcaa tctttgctat aaaaaatggc ccaaaatctc acattggaag acatttgatg
     4861 acctcatttc tttcaatgaa gggcctaacg gagttgacta atgttgtggg aaattggagc
     4921 gataagcgtg cttctgccgt ggccaggaca acgtatactc atcagataac agcaatacct
     4981 gatcactact tcgcactagt ttctcggtac tatgcatatg atccaatatc aaaggaaatg
     5041 atagcattga aggatgagac taatccaatt gaggagtggc agcatataga acagctaaag
     5101 ggtagtgctg aaggaagcat acgatacccc gcatggaatg ggataatatc acaggaggta
     5161 ctagactacc tttcatccta cataaataga cgcatataag tacgcattta agcataaaca
     5221 cgcactatgc cgttcttctc atgtatatat atatacaggc aacacgcaga tataggtgcg
     5281 acgtgaacag tgagctgtat gtgcgcagct cgcgttgcat tttcggaagc gctcgttttc
     5341 ggaaacgctt tgaagttcct attccgaagt tcctattctc tagaaagtat aggaacttca
     5401 gagcgctttt gaaaaccaaa agcgctctga agacgcactt tcaaaaaacc aaaaacgcac
     5461 cggactgtaa cgagctacta aaatattgcg aataccgctt ccacaaacat tgctcaaaag
     5521 tatctctttg ctatatatct ctgtgctata tccctatata acctacccat ccacctttcg
     5581 ctccttgaac ttgcatctaa actcgacctc tacatttttt atgtttatct ctagtattac
     5641 tctttagaca aaaaaattgt agtaagaact attcatagag tgaatcgaaa acaatacgaa
     5701 aatgtaaaca tttcctatac gtagtatata gagacaaaat agaagaaacc gttcataatt
     5761 ttctgaccaa tgaagaatca tcaacgctat cactttctgt tcacaaagta tgcgcaatcc
     5821 acatcggtat agaatataat cggggatgcc tttatcttga aaaaatgcac ccgcagcttc
     5881 gctagtaatc agtaaacgcg ggaagtggag tcaggctttt tttatggaag agaaaataga
     5941 caccaaagta gccttcttct aaccttaacg gacctacagt gcaaaaagtt atcaagagac
     6001 tgcattatag agcgcacaaa ggagaaaaaa agtaatctaa gatgctttgt tagaaaaata
     6061 gcgctctcgg gatgcatttt tgtagaacaa aaaagaagta tagattcttt gttggtaaaa
     6121 tagcgctctc gcgttgcatt tctgttctgt aaaaatgcag ctcagattct ttgtttgaaa
     6181 aattagcgct ctcgcgttgc atttttgttt tacaaaaatg aagcacagat tcttcgttgg
     6241 taaaatagcg ctttcgcgtt gcatttctgt tctgtaaaaa tgcagctcag attctttgtt
     6301 tgaaaaatta gcgctctcgc gttgcatttt tgttctacaa aatgaagcac agatgcttcg
     6361 ttaacaaaga tatgctattg aagtgcaaga tggaaacgca gaaaatgaac cggggatgcg
     6421 acgtgcaaga ttacctatgc aatagatgca atagtttctc caggaaccga aatacataca
     6481 ttgtcttccg taaagcgcta gactatatat tattatacag gttcaaatat actatctgtt
     6541 tcagggaaaa ctcccaggtt cggatgttca aaattcaatg atgggtaaca agtacgatcg
     6601 taaatctgta aaacagtttg tcggatatta ggctgtatct cctcaaagcg tattcgaata
     6661 tcattgagaa gctgcagcgt cacatcggat aataatgatg gcagccattg tagaagtgcc
     6721 ttttgcattt ctagtctctt tctcggtcta gctagtttta ctacatcgcg aagatagaat
     6781 cttagatcac actgcctttg ctgagctgga tcaatagagt aacaaaagag tggtaaggaa
     6841 ctgtgggaat actcaggtat cgtaagatgc aagagttcga atctcttagc aaccattatt
     6901 tttttcctca acataacgag aacacacagg ggcgctatcg cacagaatca aattcgatga
     6961 ctggaaattt tttgttaatt tcagaggtcg cctgacgcat ataccttttt caactgaaaa
     7021 attgggagaa aaaggaaagg tgagagcgcc ggaaccggct tttcatatag aatagagaag
     7081 cgttcatgac taaatgcttg catcacaata cttgaagttg acaatattat ttaaggacct
     7141 attgtttttt ccaataggtg gttagcaatc gtcttacttt ctaacttttc ttacctttta
     7201 catttcagca atatatatat atatttcaag gatataccat tctaatgtct gcccctaaga
     7261 agatcgtcgt tttgccaggt gaccacgttg gtcaagaaat cacagccgaa gccattaagg
     7321 ttcttaaagc tatttctgat gttcgttcca atgtcaagtt cgatttcgaa aatcatttaa
     7381 ttggtggtgc tgctatcgat gctacaggtg ttccacttcc agatgaggcg ctggaagcct
     7441 ccaagaaggc tgatgccgtt ttgttaggtg ctgtgggtgg tcctaaatgg ggtacaggta
     7501 gtgttagacc tgaacaaggt ttactaaaaa tccgtaaaga acttcaattg tacgccaact
     7561 taagaccatg taactttgca tccgactctc ttttagactt atctccaatc aagccacaat
     7621 ttgctaaagg tactgacttc gttgttgtca gagaattagt gggaggtatt tactttggta
     7681 agagaaagga agacgatggt gatggtgtcg cttgggatag tgaacaatac accgttccag
     7741 aagtgcaaag aatcacaaga atggccgctt tcatggccct acaacatgag ccaccattgc
     7801 ctatttggtc cttggataaa gctaatgttt tggcctcttc aagattatgg agaaaaactg
     7861 tggaggaaac catcaagaac gaatttccta cattgaaggt tcaacatcaa ttgattgatt
     7921 ctgccgccat gatcctagtt aagaacccaa cccacctaaa tggtattata atcaccagca
     7981 acatgtttgg tgatatcatc tccgatgaag cctccgttat cccaggttcc ttgggtttgt
     8041 tgccatctgc gtccttggcc tctttgccag acaagaacac cgcatttggt ttgtacgaac
     8101 catgccacgg ttctgctcca gatttgccaa agaataaggt caaccctatc gccactatct
     8161 tgtctgctgc aatgatgttg aaattgtcat tgaacttgcc tgaagaaggt aaggccattg
     8221 aagatgcagt taaaaaggtt ttggatgcag gtatcagaac tggtgattta ggtggttcca
     8281 acagtaccac cgaagtcggt gatgctgtcg ccgaagaagt taagaaaatc cttgcttaaa
     8341 aagattctct ttttttatga tatttgtaca taaactttat aaatgaaatt cataatagaa
     8401 acgacacgaa attacaaaat ggaatatgtt catagggtag acgaaactat atacgcaatc
     8461 tacatacatt tatcaagaag gagaaaaagg aggatgtaaa ggaatacagg taagcaaatt
     8521 gatactaatg gctcaacgtg ataaggaaaa agaattgcac tttaacatta atattgacaa
     8581 ggaggagggc accacacaaa aagttaggtg taacagaaaa tcatgaaact atgattccta
     8641 atttatatat tggaggattt tctctaaaaa aaaaaaaata caacaaataa aaaacactca
     8701 atgacctgac catttgatgg agtttaagtc aatacc
//
