LOCUS       sequence-13826-e        7771 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 7771)
  TITLE     Koloteva-Levine et al
REFERENCE   2  (bases 1 to 7771)
  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: pTH342-SUP45(276-437)-GalBD
FEATURES             Location/Qualifiers
     source          1..7771
                     /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      1805..1852
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1879..2066
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1934..1958)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2090..2106)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2090..2106)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2103..2125)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2114..2130
                     /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(2138..2144,2145..2162,2163..2168))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(2321..2338)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2492..3080)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2572..2591)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3409..3456
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3481..3500
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3689..4483)
                     /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(3800..3819)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4410..4429
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4484..4588)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4615..5957
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5578..5625)
                     /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     5998..6016
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6054..6076)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6176..6195
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6216..6497
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6498..7172
                     /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(7275..7730)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7412..7433)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7624..7643
                     /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 tggaggtcaa gtatgttcaa gaaaagaaat
     1321 tattggaggc atattttgac gaaatttccc aggacactgg taaattctgt tatggtatag
     1381 atgatacttt aaaggcattg gatttaggtg cagtcgaaaa attaattgtt ttcgaaaatt
     1441 tggaaactat cagatataca tttaaagatg ccgaggataa tgaggttata aaattcgctg
     1501 aaccagaagc caaggacaag tcgtttgcta ttgacaaagc taccggccaa gaaatggacg
     1561 ttgtctccga agaaccttta attgaatggc tagcagctaa ctacaaaaac ttcggtgcta
     1621 ccttggaatt catcacagac aaatcttcag aaggtgccca atttgtcaca ggttttggtg
     1681 gtattggtgc catgctgcgt tacaaagtta attttgaaca actagttgat gaatctgagg
     1741 atgaatatta tgacgaagat gaaggatccg actatgattt catttaactg cagcggccgc
     1801 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgcgcgcttg
     1861 cagccaagct aattccgggc gaatttctta tgatttatga tttttattat taaataagtt
     1921 ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc
     1981 ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gcatgaggtc
     2041 gctcttattg accacacctc taccggcatg caagcttggc gtaatcatgg tcatagctgt
     2101 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     2161 agtgtaaagc ctggggtgcc taatgagtga ggtaactcac attaattgcg ttgcgctcac
     2221 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2281 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2341 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2401 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2461 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2521 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2581 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2641 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2701 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2761 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2821 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2881 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     2941 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3001 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3061 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3121 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3181 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg
     3241 aggctatggc agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac
     3301 ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg
     3361 gtggggtatc gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc
     3421 caacaccgtg cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta
     3481 ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac tccagcatga
     3541 gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc
     3601 tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt
     3661 cggttcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg
     3721 cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt
     3781 cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg
     3841 ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat
     3901 tcggcaagca ggcatcgtca tgggtcacga cgagatcctc gccgtcgggc atgctcgcct
     3961 tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct
     4021 gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt
     4081 ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga
     4141 tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc
     4201 ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa
     4261 cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac
     4321 cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg
     4381 cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc
     4441 aagcggccgg agaacctgcg tgcaatccat cttgttcaat catactcttc ctttttcaat
     4501 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4561 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgaacgaa
     4621 gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca
     4681 aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct attttaccaa
     4741 cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc
     4801 aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga gcgctatttt
     4861 accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga gagcgctatt
     4921 tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat aatgcagtct
     4981 cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact ttggtgtcta
     5041 ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt actagcgaag
     5101 ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac cgatgtggat
     5161 tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg tcagaaaatt
     5221 atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt tacattttcg
     5281 tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc taaagagtaa
     5341 tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc aaggagcgaa
     5401 aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa gagatacttt
     5461 tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac agtccggtgc
     5521 gtttttggtt ttttgaaagt gcgtcttcag agcgcttttg gttttcaaaa gcgctctgaa
     5581 gttcctatac tttctagaga ataggaactt cggaatagga acttcaaagc gtttccgaaa
     5641 acgagcgctt ccgaaaatgc aacgcgagct gcgcacatac agctcactgt tcacgtcgca
     5701 cctatatctg cgtgttgcct gtatatatat atacatgaga agaacggcat agtgcgtgtt
     5761 tatgcttaaa tgcgtactta tatgcgtcta tttatgtagg atgaaaggta gtctagtacc
     5821 tcctgtgata ttatcccatt ccatgcgggg tatcgtatgc ttccttcagc actacccttt
     5881 agctgttcta tatgctgcca ctcctcaatt ggattagtct catccttcaa tgctatcatt
     5941 tcctttgata ttggatcata ttaagaaacc attattatca tgacattaac ctataaaaat
     6001 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     6061 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     6121 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     6181 tcagagcaga ttgtactgag agtgcaccat agatcaacga cattactata tatataatat
     6241 aggaagcatt taatagaaca gcatcgtaat atatgtgtac tttgcagtta tgacgccaga
     6301 tggcagtagt ggaagatatt ctttattgaa aaatagcttg tcaccttacg tacaatcttg
     6361 atccggagct tttctttttt tgccgattaa gaattaattc ggtcgaaaaa agaaaaggag
     6421 agggccaaga gggagggcat tggtgactat tgagcacgtg agtatacgtg attaagcaca
     6481 caaaggcagc ttggagtatg tctgttatta atttcacagg tagttctggt ccattggtga
     6541 aagtttgcgg cttgcagagc acagaggccg cagaatgtgc tctagattcc gatgctgact
     6601 tgctgggtat tatatgtgtg cccaatagaa agagaacaat tgacccggtt attgcaagga
     6661 aaatttcaag tcttgtaaaa gcatataaaa atagttcagg cactccgaaa tacttggttg
     6721 gcgtgtttcg taatcaacct aaggaggatg ttttggctct ggtcaatgat tacggcattg
     6781 atatcgtcca actgcatgga gatgagtcgt ggcaagaata ccaagagttc ctcggtttgc
     6841 cagttattaa aagactcgta tttccaaaag actgcaacat actactcagt gcagcttcac
     6901 agaaacctca ttcgtttatt cccttgtttg attcagaagc aggtgggaca ggtgaacttt
     6961 tggattggaa ctcgatttct gactgggttg gaaggcaaga gagccccgaa agcttacatt
     7021 ttatgttagc tggtggactg acgccagaaa atgttggtga tgcgcttaga ttaaatggcg
     7081 ttattggtgt tgatgtaagc ggaggtgtgg agacaaatgg tgtaaaagac tctaacaaaa
     7141 tagcaaattt cgtcaaaaat gctaagaaat aggttattac tgagtagtat ttatttaagt
     7201 attgtttgtg cacttgccga tctatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     7261 aataccgcat caggaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg
     7321 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     7381 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     7441 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     7501 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     7561 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     7621 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     7681 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca
     7741 ttcaggctgc gcaactgttg ggaagggcga t
//
