LOCUS       sequence-74637-e        7897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses GAL4BD-Rab30Q68LdeltaCCNFN in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7897)
  TITLE     Scidmore Lab GAL4BD plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7897)
  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            "Unpublished"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: GBK-Rab30Q68LdeltaCCNFN
FEATURES             Location/Qualifiers
     source          1..7897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..734
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             762..1202
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     primer_bind     1213..1232
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1213..1231
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1251..1280
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      1931..1978
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2005..2192
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2060..2084)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2216..2232)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2216..2232)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2229..2251)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2240..2256
                     /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(join(2264..2270,2271..2288,2289..2294))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(2447..2464)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2618..3206)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2698..2717)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3535..3582
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3607..3626
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3815..4609)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHDDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3926..3945)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4536..4555
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4610..4714)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4741..6083
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5704..5751)
                     /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)."
     primer_bind     6124..6142
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6180..6202)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6302..6321
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6342..6623
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6624..7298
                     /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"
     rep_origin      complement(7401..7856)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7538..7559)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7750..7769
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cggtgcgggc ctcttcgcta ttacgccaga tccttttgtt gtttccgggt gtacaatatg
       61 gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata ccttcgttgg
      121 tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac cagacaagac
      181 ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg tacataacga
      241 actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca ctaccctttt
      301 tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct ttttttttct
      361 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaaa tgatggaaga
      421 cactaaagga aaaaattaac gacaaagaca gcaccaacag atgtcgttgt tccagagctg
      481 atgaggggta tctcgaagca cacgaaactt tttccttcct tcattcacgc acactactct
      541 ctaatgagca acggtatacg gccttccttc cagttacttg aatttgaaat aaaaaaagtt
      601 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca
      661 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa
      721 gcatacaatc aactccaagc ttgaagcaag cctcctgaaa gatgaagcta ctgtcttcta
      781 tcgaacaagc atgcgatatt tgccgactta aaaagctcaa gtgctccaaa gaaaaaccga
      841 agtgcgccaa gtgtctgaag aacaactggg agtgtcgcta ctctcccaaa accaaaaggt
      901 ctccgctgac tagggcacat ctgacagaag tggaatcaag gctagaaaga ctggaacagc
      961 tatttctact gatttttcct cgagaagacc ttgacatgat tttgaaaatg gattctttac
     1021 aggatataaa agcattgtta acaggattat ttgtacaaga taatgtgaat aaagatgccg
     1081 tcacagatag attggcttca gtggagactg atatgcctct aacattgaga cagcatagaa
     1141 taagtgcgac atcatcatcg gaagagagta gtaacaaagg tcaaagacag ttgactgtat
     1201 cgccggaatt tgtaatacga ctcactatag ggcgagccgc catcatggag gagcagaagc
     1261 tgatctcaga ggaggacctg catatggcca tggaggccga attcccgggg atccatatga
     1321 gtatggaaga ttatgatttc ctgttcaaaa ttgttttaat tggcaacgct ggtgtgggga
     1381 agacgtgcct cgtccgaaga ttcactcagg gtcttttccc cccaggtcaa ggagccacaa
     1441 ttggagttga ttttatgatt aagacagtgg agattaatgg tgaaaaagta aagctacaga
     1501 tctgggacac agcaggtcta gagagatttc ggtccattac ccagagttac taccgaagcg
     1561 ccaatgcctt gatcctcacc tatgacatta cctgtgagga atccttccgt tgccttcctg
     1621 agtggctgcg ggagatagaa caatatgcca gcaacaaggt catcactgtg ttagtgggca
     1681 acaagattga cctggctgaa aggagagagg tttcccagca gcgagctgaa gaattctcag
     1741 aagctcagga catgtattat ctggagacct cagccaagga atctgataat gtggagaaac
     1801 tcttccttga cttagcatgc cgactcatca gtgaagccag acagaacaca cttgtgaaca
     1861 atgtatcctc acccttacct ggagaaggga aaagcatcag ctatttgact taactgcagc
     1921 ggccgcataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgcg
     1981 cgcttgcagc caagctaatt ccgggcgaat ttcttatgat ttatgatttt tattattaaa
     2041 taagttataa aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga
     2101 aaattcttat tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagcat
     2161 gaggtcgctc ttattgacca cacctctacc ggcatgcaag cttggcgtaa tcatggtcat
     2221 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     2281 gcataaagtg taaagcctgg ggtgcctaat gagtgaggta actcacatta attgcgttgc
     2341 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     2401 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2461 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2521 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2581 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2641 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2701 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2761 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2821 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2881 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     2941 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3001 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     3061 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3121 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3181 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3241 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3301 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3361 aacctgaggc tatggcaggg cctgccgccc cgacgttggc tgcgagccct gggccttcac
     3421 ccgaacttgg ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg ccccaatggg
     3481 gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt ttatgaacaa
     3541 acgacccaac accgtgcgtt ttattctgtc tttttattgc cgtcatagcg cgggttcctt
     3601 ccggtattgt ctccttccgt gtttcagtta gcctccccct agggtgggcg aagaactcca
     3661 gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg attccgaagc
     3721 ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg
     3781 cttggtcggt tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat
     3841 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
     3901 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
     3961 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
     4021 tgatattcgg caagcaggca tcgtcatggg tcacgacgag atcctcgccg tcgggcatgc
     4081 tcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
     4141 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
     4201 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
     4261 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
     4321 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
     4381 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc agttcattca
     4441 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
     4501 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
     4561 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcata ctcttccttt
     4621 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4681 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4741 aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt
     4801 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt
     4861 taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa
     4921 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc
     4981 tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc
     5041 gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg
     5101 cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg
     5161 tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta
     5221 gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat
     5281 gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag
     5341 aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca
     5401 ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa
     5461 gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg
     5521 agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga
     5581 tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc
     5641 cggtgcgttt ttggtttttt gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc
     5701 tctgaagttc ctatactttc tagagaatag gaacttcgga ataggaactt caaagcgttt
     5761 ccgaaaacga gcgcttccga aaatgcaacg cgagctgcgc acatacagct cactgttcac
     5821 gtcgcaccta tatctgcgtg ttgcctgtat atatatatac atgagaagaa cggcatagtg
     5881 cgtgtttatg cttaaatgcg tacttatatg cgtctattta tgtaggatga aaggtagtct
     5941 agtacctcct gtgatattat cccattccat gcggggtatc gtatgcttcc ttcagcacta
     6001 ccctttagct gttctatatg ctgccactcc tcaattggat tagtctcatc cttcaatgct
     6061 atcatttcct ttgatattgg atcatattaa gaaaccatta ttatcatgac attaacctat
     6121 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     6181 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     6241 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     6301 gcggcatcag agcagattgt actgagagtg caccatagat caacgacatt actatatata
     6361 taatatagga agcatttaat agaacagcat cgtaatatat gtgtactttg cagttatgac
     6421 gccagatggc agtagtggaa gatattcttt attgaaaaat agcttgtcac cttacgtaca
     6481 atcttgatcc ggagcttttc tttttttgcc gattaagaat taattcggtc gaaaaaagaa
     6541 aaggagaggg ccaagaggga gggcattggt gactattgag cacgtgagta tacgtgatta
     6601 agcacacaaa ggcagcttgg agtatgtctg ttattaattt cacaggtagt tctggtccat
     6661 tggtgaaagt ttgcggcttg cagagcacag aggccgcaga atgtgctcta gattccgatg
     6721 ctgacttgct gggtattata tgtgtgccca atagaaagag aacaattgac ccggttattg
     6781 caaggaaaat ttcaagtctt gtaaaagcat ataaaaatag ttcaggcact ccgaaatact
     6841 tggttggcgt gtttcgtaat caacctaagg aggatgtttt ggctctggtc aatgattacg
     6901 gcattgatat cgtccaactg catggagatg agtcgtggca agaataccaa gagttcctcg
     6961 gtttgccagt tattaaaaga ctcgtatttc caaaagactg caacatacta ctcagtgcag
     7021 cttcacagaa acctcattcg tttattccct tgtttgattc agaagcaggt gggacaggtg
     7081 aacttttgga ttggaactcg atttctgact gggttggaag gcaagagagc cccgaaagct
     7141 tacattttat gttagctggt ggactgacgc cagaaaatgt tggtgatgcg cttagattaa
     7201 atggcgttat tggtgttgat gtaagcggag gtgtggagac aaatggtgta aaagactcta
     7261 acaaaatagc aaatttcgtc aaaaatgcta agaaataggt tattactgag tagtatttat
     7321 ttaagtattg tttgtgcact tgccgatcta tgcggtgtga aataccgcac agatgcgtaa
     7381 ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa
     7441 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     7501 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     7561 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     7621 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     7681 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     7741 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     7801 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat
     7861 tcgccattca ggctgcgcaa ctgttgggaa gggcgat
//
