LOCUS       sequence-310760-        7685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast vector for expressing mCherry tagged genes under the GAL1 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7685)
  AUTHORS   Roy B, Granas D, Bragg F Jr, Cher JAY, White MA, Stormo GD
  TITLE     Autoregulation of yeast ribosomal proteins discovered by efficient 
            search for feedback regulation.
  JOURNAL   Commun Biol. 2020 Dec 11;3(1):761. doi: 10.1038/s42003-020-01494-z.
  PUBMED    33311538
REFERENCE   2  (bases 1 to 7685)
  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: "Commun 
            Biol."; date: "2020-12-11"; volume: "3(1):761. doi"; pages: " 
            10.1038/s42003-020-01494-z"
COMMENT     Sequence Label: pMJB1
FEATURES             Location/Qualifiers
     source          1..7685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        190..410
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             411..1214
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1343..1798)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1480..1501)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1692..1711
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1928..1950
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1942..1959
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1943..1959
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1966..1985
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1966..1984
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2002..2443
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2002..2119
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2414..2437
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(3927..3945)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4195..4214
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4673..4699
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(4911..4929)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4950..4966)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4950..4966)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4963..4985)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4974..4990
                     /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(4998..5004,5005..5022,5023..5028))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5181..5198)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5352..5940)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6111..6902,6903..6971))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6734..6753
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6972..7076)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7113..7616
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     7658..7676
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accacgcttt tcaattcaat tcatcatttt ttttttattc ttttttttga tttcggtttc
      241 tttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
      301 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
      361 tcttaaccca actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag
      421 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
      481 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
      541 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
      601 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
      661 acaatttttt actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc
      721 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
      781 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
      841 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
      901 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
      961 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     1021 tgggtttaga tgacaaggga gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     1081 tctctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     1141 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     1201 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     1261 gagcttcaat ttaattatat cagttattac cctgcggtgt gaaataccgc acagatgcgt
     1321 aaggagaaaa taccgcatca ggaaattgta aacgttaata ttttgttaaa attcgcgtta
     1381 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     1441 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     1501 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     1561 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     1621 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     1681 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     1741 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg
     1801 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     1861 ctattacgcc agctggcgaa ggggggatgt gctgcaaggc gattaagttg ggtaacgcca
     1921 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta
     1981 tagggcgaat tggcggccgc acggattaga agccgccgag cgggtgacag ccctccgaag
     2041 gaagactctc ctccgtgcgt cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc
     2101 ctcgcgccgc actgctccga acaataaaga ttctacaata ctagctttta tggttatgaa
     2161 gaggaaaaat tggcagtaac ctggccccac aaaccttcaa attaacgaat caaattaaca
     2221 accataggat gataatgcga ttagtttttt agccttattt ctggggtaat taatcagcga
     2281 agcgatgatt tttgatctat taacagatat ataaatggaa aagctgcata accactttaa
     2341 ctaatacttt caacattttc agtttgtatt acttcttatt caaatgtcat aaaagtatca
     2401 acaaaaaatt gttaatatac ctctatactt taacgtcaag gagaaggtta attaaggaga
     2461 aaaaactata gctagcatgc ttaagggccc gcttaagggc tgcttgaata tgtcgaagaa
     2521 agtgattgta atagccggaa ctacaggtgt tggcaaaagc caattatcta tacagttggc
     2581 acaaaagttt aatggcgaag taattaactc tgattccatg caagtttata aagatatacc
     2641 gataatcact aataaacatc cgctacaaga gagagaaggg attcctcatc atgtcatgaa
     2701 tcacgtggac tggtcagaag agtactattc gcataggttc gaaacggaat gcatgaacgc
     2761 tatagaagac attcatagaa ggggcaaaat acctattgtt gttggaggta cacactacta
     2821 tttacagact ctctttaaca agcgagtaga tactaaatct tccgaacgta aactaactcg
     2881 taaacagttg gatattcttg aatctacaga tcctgatgtc atctacaata ctttggtaaa
     2941 atgtgacccc gacattgcga cgaaatacca tccgaacgac tatcgtaggg tgcaaagaat
     3001 gcttgagatt tattataaga cgggcaaaaa accatctgaa acgtttaacg agcaaaagat
     3061 cactttgaaa tttgacactc ttttcttgtg gctctattct aaacccgaac cactttttca
     3121 aaggctggat gacagggtag atgatatgtt agaaagaggg gctttgcaag aaataaaaca
     3181 attgtacgaa tactacagcc aaaataaatt cacacctgaa caatgtgaga atggcgtttg
     3241 gcaggtaatt ggcttcaagg aatttctgcc ttggctgaca ggtaagacag atgataacac
     3301 tgtaaaacta gaagattgta tcgagaggat gaaaacccga acccgtcaat atgctaagcg
     3361 tcaagtaaag tggattaaaa aaatgttaat accggatata aaaggtgata tctacctatt
     3421 ggatgctaca gatctaagtc aatgggacac caatgcatcc caaagggcta tagccattag
     3481 taacgacttt atcagcaatc ggcctatcaa gcaagaaaga gctcctaagg cattggaaga
     3541 attactttca aagggtgaaa caactatgaa aaaattggat gattggactc actatacgtg
     3601 caacgtatgc agaaatgccg acggtaaaaa tgttgttgca attggagaaa agtattggaa
     3661 aatacatcta ggttcccgtc ggcataaatc caacttgaag agaaacacaa gacaggcaga
     3721 ctttgagaag tggaaaatta ataaaaaaga gacggtggag gtgagcaagg gcgaggagcc
     3781 tagggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgagattcaa
     3841 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     3901 cagaccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     3961 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     4021 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagag
     4081 agtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     4141 cggcgagttc atctacaagg tgaagttgag aggcaccaac ttcccctccg acggccccgt
     4201 aatgcagaag aagaccatgg gctgggaggc ctcctccgag agaatgtacc ccgaggacgg
     4261 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     4321 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     4381 caacatcaag ttggacatca cctcccacaa cgaggactac accattgtgg aacaatatga
     4441 aagagctgaa ggtagacatt ctactggtgg tatggatgaa ttgtacaaat aagctagcat
     4501 agattaattt aaacagtata tgtacagttt tatatatata tatatatata tatacatata
     4561 taaagaaacc tgtgcgtttt ttgtattttc aaatacattt agttttggcg ccacttctat
     4621 aaaaggtctc atgatattgt tacagatctg tttagcttgc ctcgaggtcg actatccata
     4681 cgatgttcct gattatgctt agtgtacatg acaccgatta tttaaagctg cagcatacga
     4741 tatatataca tgtgtatata tgtataccta tgaatgtcag taagtatgta tacgaacagt
     4801 atgatactga agatgacaag gtaatgcatc attctatacg tgtcattctg aacgaggcgc
     4861 gcggatccca gcttttgttc cctttagtga gggttaatca gcttttgttc cctttagtga
     4921 gggttaattc cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4981 ccgctcacaa ttccacacaa cataggagcc ggaagcataa agtgtaaagc ctggggtgcc
     5041 taatgagtga ggtaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5101 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5161 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5221 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5281 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5341 ttgctggcgt ttttccatag gctcggcccc cctgacgagc atcacaaaaa tcgacgctca
     5401 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgttccc ccctggaagc
     5461 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5521 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     5581 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5641 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5701 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5761 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     5821 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5881 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5941 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6001 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6061 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6121 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6181 ctgcccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6241 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6301 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6361 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6421 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6481 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gaaaaaaagc ggttagctcc
     6541 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6601 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6661 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6721 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6781 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6841 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6901 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6961 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7021 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7081 tttccccgaa aagtgccacc tgggtccttt tcatcacgtg ctataaaaat aattataatt
     7141 taaatttttt aatataaata tataaattaa aaatagaaag taaaaaaaga aattaaagaa
     7201 aaaatagttt ttgttttccg aagatgtaaa agactctagg gggatcgcca acaaatacta
     7261 ccttttatct tgctcttcct gctctcaggt attaatgccg aattgtttca tcttgtctgt
     7321 gtagaagacc acacacgaaa atcctgtgat tttacatttt acttatcgtt aatcgaatgt
     7381 atatctattt aatctgcttt tcttgtctaa taaatatata tgtaaagtac gctttttgtt
     7441 gaaatttttt aaacctttgt ttattttttt ttcttcattc cgtaactctt ctaccttctt
     7501 tatttacttt ctaaaatcca aatacaaaac ataaaaataa ataaacacag agtaaattcc
     7561 caaattattc catcattaaa agatacgagg cgcgtgtaag ttacaggcaa gcgatccgtc
     7621 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     7681 tcgtc
//
