LOCUS       sequence-74633-e        7933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses GAL4BD-Rab14S25N-Delta-CGC in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7933)
  TITLE     Scidmore Lab GAL4BD plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7933)
  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            "Unpublished"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-Rab14S25N-Delta-CGC
FEATURES             Location/Qualifiers
     source          1..7933
                     /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      1967..2014
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2041..2228
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2096..2120)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2252..2268)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2252..2268)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2265..2287)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2276..2292
                     /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(2300..2306,2307..2324,2325..2330))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(2483..2500)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2654..3242)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2734..2753)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3571..3618
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3643..3662
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3851..4645)
                     /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(3962..3981)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4572..4591
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4646..4750)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4777..6119
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5740..5787)
                     /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     6160..6178
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6216..6238)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6338..6357
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6378..6659
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6660..7334
                     /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(7437..7892)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7574..7595)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7786..7805
                     /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 attcatggca actacaccgt
     1321 acaactactc ttacatcttt aaatatatta ttattgggga catgggagta ggaaaaaatt
     1381 gcttgcttca tcaatttaca gaaaaaaaat ttatggctga ttgtcctcac acaattggtg
     1441 ttgaatttgg tacaagaata atcgaagtta gtggccaaaa aataaaactg cagatttggg
     1501 atacggcagg acaggagcga tttagggctg ttacacggag ctactacaga ggagctgcgg
     1561 gagctcttat ggtctatgat atcactagaa gaagtacata taaccactta agcagctggt
     1621 tgacagatgc aaggaatctc accaatccaa atactgtaat aattctcata ggaaataaag
     1681 cagatttgga ggcacagaga gatgttacat atgaagaagc caaacagttt gctgaagaaa
     1741 atggcttatt gttcctcgaa gcgagtgcaa aaacgggaga gaatgtagaa gatgccttcc
     1801 ttgaggctgc caagaaaatc tatcagaaca ttcaggatgg aagcttggat ctgaatgctg
     1861 ctgagtctgg tgtacaacac aaaccttcag ccccgcaggg aggccggcta accagtgaac
     1921 cccaacccca gagagaaggc tagctcgacc tgcagcggcc gcataactag cataacccct
     1981 tggggcctct aaacgggtct tgaggggttt tttgcgcgct tgcagccaag ctaattccgg
     2041 gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa aataagtgta
     2101 tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt gagtaactct
     2161 ttcctgtagg tcaggttgct ttctcaggta tagcatgagg tcgctcttat tgaccacacc
     2221 tctaccggca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     2281 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     2341 cctaatgagt gaggtaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2401 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2461 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2521 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2581 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2641 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2701 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2761 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2821 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2881 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2941 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3001 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3061 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     3121 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3181 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3241 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3301 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3361 aatgaagttt taaatcaatc taaagtatat atgagtaacc tgaggctatg gcagggcctg
     3421 ccgccccgac gttggctgcg agccctgggc cttcacccga acttgggggg tggggtgggg
     3481 aaaaggaaga aacgcgggcg tattggcccc aatggggtct cggtggggta tcgacagagt
     3541 gccagccctg ggaccgaacc ccgcgtttat gaacaaacga cccaacaccg tgcgttttat
     3601 tctgtctttt tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt
     3661 cagttagcct ccccctaggg tgggcgaaga actccagcat gagatccccg cgctggagga
     3721 tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg
     3781 tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg gtcggttcat ttcgaacccc
     3841 agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     3901 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     3961 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     4021 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgt
     4081 catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt
     4141 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     4201 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     4261 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     4321 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     4381 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     4441 acgatagccg cgctgcctcg tcttgcagtt cattcagggc accggacagg tcggtcttga
     4501 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     4561 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     4621 cgtgcaatcc atcttgttca atcatactct tcctttttca atattattga agcatttatc
     4681 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4741 gggttccgcg cacatttccc cgaaaagtgc cacctgaacg aagcatctgt gcttcatttt
     4801 gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     4861 tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca
     4921 tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     4981 gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata
     5041 cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt
     5101 agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac
     5161 tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa
     5221 aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc
     5281 aagataaagg catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac
     5341 agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat
     5401 tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact
     5461 ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa
     5521 aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta
     5581 tatagggata tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa
     5641 gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa
     5701 gtgcgtcttc agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctaga
     5761 gaataggaac ttcggaatag gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat
     5821 gcaacgcgag ctgcgcacat acagctcact gttcacgtcg cacctatatc tgcgtgttgc
     5881 ctgtatatat atatacatga gaagaacggc atagtgcgtg tttatgctta aatgcgtact
     5941 tatatgcgtc tatttatgta ggatgaaagg tagtctagta cctcctgtga tattatccca
     6001 ttccatgcgg ggtatcgtat gcttccttca gcactaccct ttagctgttc tatatgctgc
     6061 cactcctcaa ttggattagt ctcatccttc aatgctatca tttcctttga tattggatca
     6121 tattaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     6181 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     6241 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     6301 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     6361 agagtgcacc atagatcaac gacattacta tatatataat ataggaagca tttaatagaa
     6421 cagcatcgta atatatgtgt actttgcagt tatgacgcca gatggcagta gtggaagata
     6481 ttctttattg aaaaatagct tgtcacctta cgtacaatct tgatccggag cttttctttt
     6541 tttgccgatt aagaattaat tcggtcgaaa aaagaaaagg agagggccaa gagggagggc
     6601 attggtgact attgagcacg tgagtatacg tgattaagca cacaaaggca gcttggagta
     6661 tgtctgttat taatttcaca ggtagttctg gtccattggt gaaagtttgc ggcttgcaga
     6721 gcacagaggc cgcagaatgt gctctagatt ccgatgctga cttgctgggt attatatgtg
     6781 tgcccaatag aaagagaaca attgacccgg ttattgcaag gaaaatttca agtcttgtaa
     6841 aagcatataa aaatagttca ggcactccga aatacttggt tggcgtgttt cgtaatcaac
     6901 ctaaggagga tgttttggct ctggtcaatg attacggcat tgatatcgtc caactgcatg
     6961 gagatgagtc gtggcaagaa taccaagagt tcctcggttt gccagttatt aaaagactcg
     7021 tatttccaaa agactgcaac atactactca gtgcagcttc acagaaacct cattcgttta
     7081 ttcccttgtt tgattcagaa gcaggtggga caggtgaact tttggattgg aactcgattt
     7141 ctgactgggt tggaaggcaa gagagccccg aaagcttaca ttttatgtta gctggtggac
     7201 tgacgccaga aaatgttggt gatgcgctta gattaaatgg cgttattggt gttgatgtaa
     7261 gcggaggtgt ggagacaaat ggtgtaaaag actctaacaa aatagcaaat ttcgtcaaaa
     7321 atgctaagaa ataggttatt actgagtagt atttatttaa gtattgtttg tgcacttgcc
     7381 gatctatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat
     7441 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     7501 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     7561 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     7621 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     7681 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     7741 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     7801 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     7861 cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt
     7921 tgggaagggc gat
//
