LOCUS       sequence-310779-        6794 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ""cre-less"" yeast RPS22B tagged with mCherry under the 
            GAL1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6794)
  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 6794)
  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-RPS22B-Cl
FEATURES             Location/Qualifiers
     source          1..6794
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(560..578)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     828..847
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1306..1332
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1544..1562)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1583..1599)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1583..1599)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1596..1618)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1607..1623
                     /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(1631..1637,1638..1655,1656..1661))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(1814..1831)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1985..2573)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(2744..3535,3536..3604))
                     /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     3367..3386
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3605..3709)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3746..4249
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4291..4309
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4347..4369)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4469..4488
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4508..4728
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4729..5532
                     /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(5661..6116)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5798..5819)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6010..6029
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6246..6268
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6260..6277
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6261..6277
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6284..6303
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6284..6302
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        6320..6761
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    6320..6437
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     6732..6755
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
ORIGIN
        1 atgactcgga gctctgtatt ggccgatgct ttgaacgcca tcaacaatgc cgaaaagacc
       61 ggtaagagac aagttttaat tagaccatcc tccaaggtta ttattaagtt cttgcaagtt
      121 atgcaaaagc acggttacat tggtgaattt gaatacattg acgaccatcg ttccggtaag
      181 atcgttgttc aattgaacgg tagattgaac aagtgtggtg ttatttctcc aagatttaac
      241 gttaagatcg gtgacattga aaaatggacc gctaacttgt tgccagctag acaattcggt
      301 tacgttatct taaccacctc cgccggtatc atggatcacg aagaagctag aagaaagcac
      361 gtctctggta agattttggg tttcgtctac taagtgagca agggcgagga gcctagggtg
      421 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgagatt caaggtgcac
      481 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggcagaccc
      541 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
      601 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
      661 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gagagtgatg
      721 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
      781 ttcatctaca aggtgaagtt gagaggcacc aacttcccct ccgacggccc cgtaatgcag
      841 aagaagacca tgggctggga ggcctcctcc gagagaatgt accccgagga cggcgccctg
      901 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
      961 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     1021 aagttggaca tcacctccca caacgaggac tacaccattg tggaacaata tgaaagagct
     1081 gaaggtagac attctactgg tggtatggat gaattgtaca aataagctag catagattaa
     1141 tttaaacagt atatgtacag ttttatatat atatatatat atatatacat atataaagaa
     1201 acctgtgcgt tttttgtatt ttcaaataca tttagttttg gcgccacttc tataaaaggt
     1261 ctcatgatat tgttacagat ctgtttagct tgcctcgagg tcgactatcc atacgatgtt
     1321 cctgattatg cttagtgtac atgacaccga ttatttaaag ctgcagcata cgatatatat
     1381 acatgtgtat atatgtatac ctatgaatgt cagtaagtat gtatacgaac agtatgatac
     1441 tgaagatgac aaggtaatgc atcattctat acgtgtcatt ctgaacgagg cgcgcggatc
     1501 ccagcttttg ttccctttag tgagggttaa tcagcttttg ttccctttag tgagggttaa
     1561 ttccgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     1621 caattccaca caacatagga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     1681 tgaggtaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     1741 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     1801 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     1861 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     1921 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1981 cgtttttcca taggctcggc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2041 ggtggcgaaa cccgacagga ctataaagat accaggcgtt cccccctgga agctccctcg
     2101 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2161 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     2221 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2281 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2341 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2401 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     2461 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2521 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2581 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2641 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2701 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     2761 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactgcccg
     2821 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     2881 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     2941 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3001 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3061 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3121 gatcaaggcg agttacatga tcccccatgt tgtgaaaaaa agcggttagc tccttcggtc
     3181 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3241 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     3301 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     3361 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     3421 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     3481 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     3541 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     3601 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     3661 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     3721 gaaaagtgcc acctgggtcc ttttcatcac gtgctataaa aataattata atttaaattt
     3781 tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag
     3841 tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta
     3901 tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag
     3961 accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta
     4021 tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt
     4081 tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac
     4141 tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta
     4201 ttccatcatt aaaagatacg aggcgcgtgt aagttacagg caagcgatcc gtcctaagaa
     4261 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     4321 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     4381 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     4441 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     4501 cacgcttttc aattcaattc atcatttttt ttttattctt ttttttgatt tcggtttctt
     4561 tgaaattttt ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga
     4621 cttagattgg tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc
     4681 ttaacccaac tgcacagaac aaaaacctgc aggaaacgaa gataaatcat gtcgaaagct
     4741 acatataagg aacgtgctgc tactcatcct agtcctgttg ctgccaagct atttaatatc
     4801 atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac caaggaatta
     4861 ctggagttag ttgaagcatt aggtcccaaa atttgtttac taaaaacaca tgtggatatc
     4921 ttgactgatt tttccatgga gggcacagtt aagccgctaa aggcattatc cgccaagtac
     4981 aattttttac tcttcgaaga cagaaaattt gctgacattg gtaatacagt caaattgcag
     5041 tactctgcgg gtgtatacag aatagcagaa tgggcagaca ttacgaatgc acacggtgtg
     5101 gtgggcccag gtattgttag cggtttgaag caggcggcag aagaagtaac aaaggaacct
     5161 agaggccttt tgatgttagc agaattgtca tgcaagggct ccctatctac tggagaatat
     5221 actaagggta ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg ctttattgct
     5281 caaagagaca tgggtggaag agatgaaggt tacgattggt tgattatgac acccggtgtg
     5341 ggtttagatg acaagggaga cgcattgggt caacagtata gaaccgtgga tgatgtggtc
     5401 tctacaggat ctgacattat tattgttgga agaggactat ttgcaaaggg aagggatgct
     5461 aaggtagagg gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc
     5521 cagcaaaact aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga
     5581 gcttcaattt aattatatca gttattaccc tgcggtgtga aataccgcac agatgcgtaa
     5641 ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa
     5701 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     5761 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     5821 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     5881 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     5941 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     6001 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     6061 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcgcg
     6121 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     6181 attacgccag ctggcgaagg ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     6241 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg actcactata
     6301 gggcgaattg gcggccgcac ggattagaag ccgccgagcg ggtgacagcc ctccgaagga
     6361 agactctcct ccgtgcgtcc tcgtcttcac cggtcgcgtt cctgaaacgc agatgtgcct
     6421 cgcgccgcac tgctccgaac aataaagatt ctacaatact agcttttatg gttatgaaga
     6481 ggaaaaattg gcagtaacct ggccccacaa accttcaaat taacgaatca aattaacaac
     6541 cataggatga taatgcgatt agttttttag ccttatttct ggggtaatta atcagcgaag
     6601 cgatgatttt tgatctatta acagatatat aaatggaaaa gctgcataac cactttaact
     6661 aatactttca acattttcag tttgtattac ttcttattca aatgtcataa aagtatcaac
     6721 aaaaaattgt taatatacct ctatacttta acgtcaagga gaaggttaat taaggagaaa
     6781 aaactatagc tagc
//
