LOCUS       sequence-13825-e        8179 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 8179)
  TITLE     Koloteva-Levine et al
REFERENCE   2  (bases 1 to 8179)
  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: pTH341-SUP45(139-437)-GalBD
FEATURES             Location/Qualifiers
     source          1..8179
                     /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      2213..2260
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2287..2474
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2342..2366)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2498..2514)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2498..2514)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2511..2533)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2522..2538
                     /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(2546..2552,2553..2570,2571..2576))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(2729..2746)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2900..3488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2980..2999)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3817..3864
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3889..3908
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4097..4891)
                     /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(4208..4227)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4818..4837
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4892..4996)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5023..6365
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5986..6033)
                     /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     6406..6424
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6462..6484)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6584..6603
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6624..6905
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6906..7580
                     /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(7683..8138)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7820..7841)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8032..8051
                     /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 tggaggctga cgacaagttc ggttttatag
     1321 tcatggacgg tcaaggtact ttgtttggtt ctgtgtccgg taatacgaga actgttttac
     1381 ataaatttac tgtcgatctg ccaaaaaagc atggtagagg tggtcaatct gcgcttcgtt
     1441 ttgctcgttt aagagaagaa aaaagacata attatgtgag aaaggtcgcc gaagttgctg
     1501 ttcaaaattt tattactaat gacaaagtca atgttaaggg tttaatttta gctggttctg
     1561 ctgactttaa gaccgatttg gctaaatctg aattattcga tccaagacta gcatgtaagg
     1621 ttatttccat cgtggatgtt tcttatggtg gtgaaaacgg tttcaaccag gctatcgaac
     1681 tttctgccga agcgttggcc aatgtcaagt atgttcaaga aaagaaatta ttggaggcat
     1741 attttgacga aatttcccag gacactggta aattctgtta tggtatagat gatactttaa
     1801 aggcattgga tttaggtgca gtcgaaaaat taattgtttt cgaaaatttg gaaactatca
     1861 gatatacatt taaagatgcc gaggataatg aggttataaa attcgctgaa ccagaagcca
     1921 aggacaagtc gtttgctatt gacaaagcta ccggccaaga aatggacgtt gtctccgaag
     1981 aacctttaat tgaatggcta gcagctaact acaaaaactt cggtgctacc ttggaattca
     2041 tcacagacaa atcttcagaa ggtgcccaat ttgtcacagg ttttggtggt attggtgcca
     2101 tgctgcgtta caaagttaat tttgaacaac tagttgatga atctgaggat gaatattatg
     2161 acgaagatga aggatccgac tatgatttca tttaactgca gcggccgcat aactagcata
     2221 accccttggg gcctctaaac gggtcttgag gggttttttg cgcgcttgca gccaagctaa
     2281 ttccgggcga atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata
     2341 agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt
     2401 aactctttcc tgtaggtcag gttgctttct caggtatagc atgaggtcgc tcttattgac
     2461 cacacctcta ccggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     2521 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     2581 ggggtgccta atgagtgagg taactcacat taattgcgtt gcgctcactg cccgctttcc
     2641 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2701 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2761 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2821 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2881 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2941 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3001 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3061 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3121 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3181 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3241 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3301 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     3361 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3421 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3481 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3541 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3601 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaacctgag gctatggcag
     3661 ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc acccgaactt ggggggtggg
     3721 gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg gggtctcggt ggggtatcga
     3781 cagagtgcca gccctgggac cgaaccccgc gtttatgaac aaacgaccca acaccgtgcg
     3841 ttttattctg tctttttatt gccgtcatag cgcgggttcc ttccggtatt gtctccttcc
     3901 gtgtttcagt tagcctcccc ctagggtggg cgaagaactc cagcatgaga tccccgcgct
     3961 ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg
     4021 cggcggtgga atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gttcatttcg
     4081 aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     4141 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
     4201 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
     4261 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     4321 catcgtcatg ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg agcctggcga
     4381 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     4441 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     4501 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     4561 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     4621 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     4681 ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg
     4741 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     4801 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     4861 aacctgcgtg caatccatct tgttcaatca tactcttcct ttttcaatat tattgaagca
     4921 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4981 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgaacgaagc atctgtgctt
     5041 cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
     5101 tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt
     5161 gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc
     5221 tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa
     5281 tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag
     5341 catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt
     5401 ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca
     5461 taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat
     5521 tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt
     5581 gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc
     5641 ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga
     5701 ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa
     5761 acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt
     5821 aggttatata gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt
     5881 gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt
     5941 ttgaaagtgc gtcttcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt
     6001 tctagagaat aggaacttcg gaataggaac ttcaaagcgt ttccgaaaac gagcgcttcc
     6061 gaaaatgcaa cgcgagctgc gcacatacag ctcactgttc acgtcgcacc tatatctgcg
     6121 tgttgcctgt atatatatat acatgagaag aacggcatag tgcgtgttta tgcttaaatg
     6181 cgtacttata tgcgtctatt tatgtaggat gaaaggtagt ctagtacctc ctgtgatatt
     6241 atcccattcc atgcggggta tcgtatgctt ccttcagcac taccctttag ctgttctata
     6301 tgctgccact cctcaattgg attagtctca tccttcaatg ctatcatttc ctttgatatt
     6361 ggatcatatt aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     6421 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     6481 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     6541 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     6601 gtactgagag tgcaccatag atcaacgaca ttactatata tataatatag gaagcattta
     6661 atagaacagc atcgtaatat atgtgtactt tgcagttatg acgccagatg gcagtagtgg
     6721 aagatattct ttattgaaaa atagcttgtc accttacgta caatcttgat ccggagcttt
     6781 tctttttttg ccgattaaga attaattcgg tcgaaaaaag aaaaggagag ggccaagagg
     6841 gagggcattg gtgactattg agcacgtgag tatacgtgat taagcacaca aaggcagctt
     6901 ggagtatgtc tgttattaat ttcacaggta gttctggtcc attggtgaaa gtttgcggct
     6961 tgcagagcac agaggccgca gaatgtgctc tagattccga tgctgacttg ctgggtatta
     7021 tatgtgtgcc caatagaaag agaacaattg acccggttat tgcaaggaaa atttcaagtc
     7081 ttgtaaaagc atataaaaat agttcaggca ctccgaaata cttggttggc gtgtttcgta
     7141 atcaacctaa ggaggatgtt ttggctctgg tcaatgatta cggcattgat atcgtccaac
     7201 tgcatggaga tgagtcgtgg caagaatacc aagagttcct cggtttgcca gttattaaaa
     7261 gactcgtatt tccaaaagac tgcaacatac tactcagtgc agcttcacag aaacctcatt
     7321 cgtttattcc cttgtttgat tcagaagcag gtgggacagg tgaacttttg gattggaact
     7381 cgatttctga ctgggttgga aggcaagaga gccccgaaag cttacatttt atgttagctg
     7441 gtggactgac gccagaaaat gttggtgatg cgcttagatt aaatggcgtt attggtgttg
     7501 atgtaagcgg aggtgtggag acaaatggtg taaaagactc taacaaaata gcaaatttcg
     7561 tcaaaaatgc taagaaatag gttattactg agtagtattt atttaagtat tgtttgtgca
     7621 cttgccgatc tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     7681 ggaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     7741 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     7801 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     7861 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     7921 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     7981 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     8041 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     8101 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc attcgccatt caggctgcgc
     8161 aactgttggg aagggcgat
//
