LOCUS       sequence-13823-e        7696 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 7696)
  TITLE     Koloteva-Levine et al
REFERENCE   2  (bases 1 to 7696)
  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: pTH339-SUP45(1-138)-GalBD
FEATURES             Location/Qualifiers
     source          1..7696
                     /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      1730..1777
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1804..1991
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1859..1883)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2015..2031)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2015..2031)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2028..2050)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2039..2055
                     /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(2063..2069,2070..2087,2088..2093))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(2246..2263)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2417..3005)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2497..2516)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3334..3381
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3406..3425
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3614..4408)
                     /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(3725..3744)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4335..4354
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4409..4513)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4540..5882
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5503..5550)
                     /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     5923..5941
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5979..6001)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6101..6120
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6141..6422
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6423..7097
                     /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(7200..7655)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7337..7358)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7549..7568
                     /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 ttgcttcaat aactgcagcg gccgcataac tagcataacc
     1741 ccttggggcc tctaaacggg tcttgagggg ttttttgcgc gcttgcagcc aagctaattc
     1801 cgggcgaatt tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt
     1861 gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac
     1921 tctttcctgt aggtcaggtt gctttctcag gtatagcatg aggtcgctct tattgaccac
     1981 acctctaccg gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     2041 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     2101 gtgcctaatg agtgaggtaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     2161 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2221 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2281 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2341 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2401 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2461 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2521 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2581 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2641 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2701 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2761 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2821 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     2881 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2941 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3001 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3061 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3121 aaaaatgaag ttttaaatca atctaaagta tatatgagta acctgaggct atggcagggc
     3181 ctgccgcccc gacgttggct gcgagccctg ggccttcacc cgaacttggg gggtggggtg
     3241 gggaaaagga agaaacgcgg gcgtattggc cccaatgggg tctcggtggg gtatcgacag
     3301 agtgccagcc ctgggaccga accccgcgtt tatgaacaaa cgacccaaca ccgtgcgttt
     3361 tattctgtct ttttattgcc gtcatagcgc gggttccttc cggtattgtc tccttccgtg
     3421 tttcagttag cctcccccta gggtgggcga agaactccag catgagatcc ccgcgctgga
     3481 ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg
     3541 cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtt catttcgaac
     3601 cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat
     3661 cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt
     3721 cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc
     3781 cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat
     3841 cgtcatgggt cacgacgaga tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca
     3901 gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg
     3961 cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg
     4021 tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg
     4081 caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt
     4141 cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca
     4201 gccacgatag ccgcgctgcc tcgtcttgca gttcattcag ggcaccggac aggtcggtct
     4261 tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc
     4321 cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac
     4381 ctgcgtgcaa tccatcttgt tcaatcatac tcttcctttt tcaatattat tgaagcattt
     4441 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4501 taggggttcc gcgcacattt ccccgaaaag tgccacctga acgaagcatc tgtgcttcat
     4561 tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     4621 atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct
     4681 tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga
     4741 gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct
     4801 atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat
     4861 cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg
     4921 cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa
     4981 aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt
     5041 ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg
     5101 aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc
     5161 tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc
     5221 actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca
     5281 taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg
     5341 ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg
     5401 gaagcggtat tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt tggttttttg
     5461 aaagtgcgtc ttcagagcgc ttttggtttt caaaagcgct ctgaagttcc tatactttct
     5521 agagaatagg aacttcggaa taggaacttc aaagcgtttc cgaaaacgag cgcttccgaa
     5581 aatgcaacgc gagctgcgca catacagctc actgttcacg tcgcacctat atctgcgtgt
     5641 tgcctgtata tatatataca tgagaagaac ggcatagtgc gtgtttatgc ttaaatgcgt
     5701 acttatatgc gtctatttat gtaggatgaa aggtagtcta gtacctcctg tgatattatc
     5761 ccattccatg cggggtatcg tatgcttcct tcagcactac cctttagctg ttctatatgc
     5821 tgccactcct caattggatt agtctcatcc ttcaatgcta tcatttcctt tgatattgga
     5881 tcatattaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     5941 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     6001 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     6061 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     6121 ctgagagtgc accatagatc aacgacatta ctatatatat aatataggaa gcatttaata
     6181 gaacagcatc gtaatatatg tgtactttgc agttatgacg ccagatggca gtagtggaag
     6241 atattcttta ttgaaaaata gcttgtcacc ttacgtacaa tcttgatccg gagcttttct
     6301 ttttttgccg attaagaatt aattcggtcg aaaaaagaaa aggagagggc caagagggag
     6361 ggcattggtg actattgagc acgtgagtat acgtgattaa gcacacaaag gcagcttgga
     6421 gtatgtctgt tattaatttc acaggtagtt ctggtccatt ggtgaaagtt tgcggcttgc
     6481 agagcacaga ggccgcagaa tgtgctctag attccgatgc tgacttgctg ggtattatat
     6541 gtgtgcccaa tagaaagaga acaattgacc cggttattgc aaggaaaatt tcaagtcttg
     6601 taaaagcata taaaaatagt tcaggcactc cgaaatactt ggttggcgtg tttcgtaatc
     6661 aacctaagga ggatgttttg gctctggtca atgattacgg cattgatatc gtccaactgc
     6721 atggagatga gtcgtggcaa gaataccaag agttcctcgg tttgccagtt attaaaagac
     6781 tcgtatttcc aaaagactgc aacatactac tcagtgcagc ttcacagaaa cctcattcgt
     6841 ttattccctt gtttgattca gaagcaggtg ggacaggtga acttttggat tggaactcga
     6901 tttctgactg ggttggaagg caagagagcc ccgaaagctt acattttatg ttagctggtg
     6961 gactgacgcc agaaaatgtt ggtgatgcgc ttagattaaa tggcgttatt ggtgttgatg
     7021 taagcggagg tgtggagaca aatggtgtaa aagactctaa caaaatagca aatttcgtca
     7081 aaaatgctaa gaaataggtt attactgagt agtatttatt taagtattgt ttgtgcactt
     7141 gccgatctat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga
     7201 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     7261 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     7321 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     7381 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     7441 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     7501 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     7561 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     7621 acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac
     7681 tgttgggaag ggcgat
//
