LOCUS       sequence-317512-        6764 bp    DNA     circular SYN 14-APR-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 6764)
  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 6764)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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..6764
                     /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(1514..1532)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1553..1569)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1553..1569)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1566..1588)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1577..1593
                     /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(1601..1607,1608..1625,1626..1631))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1646..1667
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1784..1801)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1955..2543)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2035..2054)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2714..3505,3506..3574))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3337..3356
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3575..3679)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3716..4219
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4260..4278
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4316..4338)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4438..4457
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4481..4696
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4697..5500
                     /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(5631..6086)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5768..5789)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5980..5999
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6216..6238
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6230..6247
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6231..6247
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6254..6273
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6254..6272
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        6290..6731
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    6290..6407
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     6702..6725
                     /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 tttcgagctt ggcgtaatca tggtcatagc
     1561 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     1621 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     1681 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     1741 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     1801 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     1861 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     1921 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1981 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2041 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2101 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2161 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2221 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2281 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2341 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2401 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2461 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2521 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2581 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     2641 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     2701 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     2761 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     2821 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     2881 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     2941 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3001 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3061 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3121 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3181 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3241 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     3301 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     3361 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     3421 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     3481 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     3541 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     3601 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     3661 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgggtcc ttttcatcac
     3721 gtgctataaa aataattata atttaaattt tttaatataa atatataaat taaaaataga
     3781 aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt ccgaagatgt aaaagactct
     3841 agggggatcg ccaacaaata ctacctttta tcttgctctt cctgctctca ggtattaatg
     3901 ccgaattgtt tcatcttgtc tgtgtagaag accacacacg aaaatcctgt gattttacat
     3961 tttacttatc gttaatcgaa tgtatatcta tttaatctgc ttttcttgtc taataaatat
     4021 atatgtaaag tacgcttttt gttgaaattt tttaaacctt tgtttatttt tttttcttca
     4081 ttccgtaact cttctacctt ctttatttac tttctaaaat ccaaatacaa aacataaaaa
     4141 taaataaaca cagagtaaat tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt
     4201 aagttacagg caagcgatcc gtctaagaaa ccattattat catgacatta acctataaaa
     4261 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     4321 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     4381 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     4441 catcagagca gattgtactg agagtgcacc ataccacctt ttcaattcat catttttttt
     4501 ttattctttt ttttgatttc ggtttccttg aaattttttt gattcggtaa tctccgaaca
     4561 gaaggaagaa cgaaggaagg agcacagact tagattggta tatatacgca tatgtagtgt
     4621 tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacctgcag
     4681 gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag
     4741 tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc
     4801 attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat
     4861 ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa
     4921 gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc
     4981 tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg
     5041 ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca
     5101 ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg
     5161 caagggctcc ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga
     5221 caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta
     5281 cgattggttg attatgacac ccggtgtggg tttagatgac aagggagacg cattgggtca
     5341 acagtataga accgtggatg atgtggtctc tacaggatct gacattatta ttgttggaag
     5401 aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg
     5461 ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa
     5521 tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccta
     5581 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag
     5641 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     5701 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     5761 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     5821 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     5881 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     5941 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     6001 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     6061 gcttaatgcg ccgctacagg gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg
     6121 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaagg ggggatgtgc
     6181 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     6241 ggccagtgaa ttgtaatacg actcactata gggcgaattg gcggccgcac ggattagaag
     6301 ccgccgagcg ggcgacagcc ctccgacgga agactctcct ccgtgcgtcc tcgtcttcac
     6361 cggtcgcgtt cctgaaacgc agatgtgcct cgcgccgcac tgctccgaac aataaagatt
     6421 ctacaatact agcttttatg gttatgaaga ggaaaaattg gcagtaacct ggccccacaa
     6481 accttcaaat taacgaatca aattaacaac cataggatga taatgcgatt agttttttag
     6541 ccttatttct ggggtaatta atcagcgaag cgatgatttt tgatctatta acagatatat
     6601 aaatggaaaa gctgcataac cactttaact aatactttca acattttcag tttgtattac
     6661 ttcttattca aatgtcataa aagtatcaac aaaaaattgt taatatacct ctatacttta
     6721 acgtcaagga gaaggttaat taaggagaaa aaactatagc tagc
//
