LOCUS       sequence-13824-e        8104 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 8104)
  TITLE     Koloteva-Levine et al
REFERENCE   2  (bases 1 to 8104)
  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: pTH340-SUP45(1-242)-GalBD
FEATURES             Location/Qualifiers
     source          1..8104
                     /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      2138..2185
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2212..2399
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2267..2291)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2423..2439)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2423..2439)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2436..2458)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2447..2463
                     /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(2471..2477,2478..2495,2496..2501))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(2654..2671)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2825..3413)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2905..2924)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3742..3789
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3814..3833
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4022..4816)
                     /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(4133..4152)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4743..4762
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4817..4921)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4948..6290
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5911..5958)
                     /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     6331..6349
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6387..6409)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6509..6528
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6549..6830
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6831..7505
                     /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(7608..8063)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7745..7766)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7957..7976
                     /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 tggagatgga taacgaggtt gaaaaaaata
     1321 ttgagatctg gaaggtcaag aagttggtcc aatctttaga aaaagctaga ggtaatggta
     1381 cttctatgat ttccttagtt attcctccta agggtcaaat tccactgtac caaaaaatgt
     1441 taacagatga atatggtact gcctcgaata ttaaatctag ggttaatcgt ctttccgttt
     1501 tatctgctat cacttccacc caacaaaagt tgaagctata taatactttg cccaagaacg
     1561 gtttagtttt atattgtggt gatatcatca ctgaagatgg taaagaaaaa aaggtcactt
     1621 ttgacatcga accttacaaa cctatcaaca catccttata tttgtgtgat aacaaatttc
     1681 atacagaagt tctttcggaa ttgcttcaag ctgacgacaa gttcggtttt atagtcatgg
     1741 acggtcaagg tactttgttt ggttctgtgt ccggtaatac gagaactgtt ttacataaat
     1801 ttactgtcga tctgccaaaa aagcatggta gaggtggtca atctgcgctt cgttttgctc
     1861 gtttaagaga agaaaaaaga cataattatg tgagaaaggt cgccgaagtt gctgttcaaa
     1921 attttattac taatgacaaa gtcaatgtta agggtttaat tttagctggt tctgctgact
     1981 ttaagaccga tttggctaaa tctgaattat tcgatccaag actagcatgt aaggttattt
     2041 ccatcgtgga tgtttcttat ggtggtgaaa acggtttcaa ccaggctatc gaactttctg
     2101 ccgaagcgtt ggccaattaa ctgcagcggc cgcataacta gcataacccc ttggggcctc
     2161 taaacgggtc ttgaggggtt ttttgcgcgc ttgcagccaa gctaattccg ggcgaatttc
     2221 ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt
     2281 taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag
     2341 gtcaggttgc tttctcaggt atagcatgag gtcgctctta ttgaccacac ctctaccggc
     2401 atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     2461 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     2521 tgaggtaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     2581 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2641 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2701 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2761 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2821 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2881 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2941 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3001 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3061 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3121 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3181 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3241 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     3301 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3361 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3421 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3481 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3541 ttaaatcaat ctaaagtata tatgagtaac ctgaggctat ggcagggcct gccgccccga
     3601 cgttggctgc gagccctggg ccttcacccg aacttggggg gtggggtggg gaaaaggaag
     3661 aaacgcgggc gtattggccc caatggggtc tcggtggggt atcgacagag tgccagccct
     3721 gggaccgaac cccgcgttta tgaacaaacg acccaacacc gtgcgtttta ttctgtcttt
     3781 ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc
     3841 tccccctagg gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc
     3901 cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga
     3961 aatctcgtga tggcaggttg ggcgtcgctt ggtcggttca tttcgaaccc cagagtcccg
     4021 ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga
     4081 taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac
     4141 gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga
     4201 atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg tcatgggtca
     4261 cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg
     4321 cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag
     4381 tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa
     4441 gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt
     4501 gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt
     4561 cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc
     4621 gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa
     4681 ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt
     4741 gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc
     4801 catcttgttc aatcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4861 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4921 gcacatttcc ccgaaaagtg ccacctgaac gaagcatctg tgcttcattt tgtagaacaa
     4981 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     5041 cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa
     5101 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     5161 cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt
     5221 tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt
     5281 tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc
     5341 gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac
     5401 tccacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag
     5461 gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga
     5521 tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct
     5581 atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata
     5641 gttcttacta caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag
     5701 aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat
     5761 atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc
     5821 gcaatatttt agtagctcgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtctt
     5881 cagagcgctt ttggttttca aaagcgctct gaagttccta tactttctag agaataggaa
     5941 cttcggaata ggaacttcaa agcgtttccg aaaacgagcg cttccgaaaa tgcaacgcga
     6001 gctgcgcaca tacagctcac tgttcacgtc gcacctatat ctgcgtgttg cctgtatata
     6061 tatatacatg agaagaacgg catagtgcgt gtttatgctt aaatgcgtac ttatatgcgt
     6121 ctatttatgt aggatgaaag gtagtctagt acctcctgtg atattatccc attccatgcg
     6181 gggtatcgta tgcttccttc agcactaccc tttagctgtt ctatatgctg ccactcctca
     6241 attggattag tctcatcctt caatgctatc atttcctttg atattggatc atattaagaa
     6301 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     6361 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     6421 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     6481 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     6541 catagatcaa cgacattact atatatataa tataggaagc atttaataga acagcatcgt
     6601 aatatatgtg tactttgcag ttatgacgcc agatggcagt agtggaagat attctttatt
     6661 gaaaaatagc ttgtcacctt acgtacaatc ttgatccgga gcttttcttt ttttgccgat
     6721 taagaattaa ttcggtcgaa aaaagaaaag gagagggcca agagggaggg cattggtgac
     6781 tattgagcac gtgagtatac gtgattaagc acacaaaggc agcttggagt atgtctgtta
     6841 ttaatttcac aggtagttct ggtccattgg tgaaagtttg cggcttgcag agcacagagg
     6901 ccgcagaatg tgctctagat tccgatgctg acttgctggg tattatatgt gtgcccaata
     6961 gaaagagaac aattgacccg gttattgcaa ggaaaatttc aagtcttgta aaagcatata
     7021 aaaatagttc aggcactccg aaatacttgg ttggcgtgtt tcgtaatcaa cctaaggagg
     7081 atgttttggc tctggtcaat gattacggca ttgatatcgt ccaactgcat ggagatgagt
     7141 cgtggcaaga ataccaagag ttcctcggtt tgccagttat taaaagactc gtatttccaa
     7201 aagactgcaa catactactc agtgcagctt cacagaaacc tcattcgttt attcccttgt
     7261 ttgattcaga agcaggtggg acaggtgaac ttttggattg gaactcgatt tctgactggg
     7321 ttggaaggca agagagcccc gaaagcttac attttatgtt agctggtgga ctgacgccag
     7381 aaaatgttgg tgatgcgctt agattaaatg gcgttattgg tgttgatgta agcggaggtg
     7441 tggagacaaa tggtgtaaaa gactctaaca aaatagcaaa tttcgtcaaa aatgctaaga
     7501 aataggttat tactgagtag tatttattta agtattgttt gtgcacttgc cgatctatgc
     7561 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaaacgt
     7621 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     7681 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     7741 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     7801 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     7861 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     7921 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     7981 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     8041 taatgcgccg ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg
     8101 cgat
//
