LOCUS       sequence-310781-        7052 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ""cre-less"" yeast RPL1B tagged with mCherry under the 
            GAL1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7052)
  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 7052)
  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-RPL1B-Cl
FEATURES             Location/Qualifiers
     source          1..7052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(818..836)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1086..1105
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1564..1590
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1802..1820)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1841..1857)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1841..1857)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1854..1876)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1865..1881
                     /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(1889..1895,1896..1913,1914..1919))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(2072..2089)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2243..2831)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(3002..3793,3794..3862))
                     /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     3625..3644
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3863..3967)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4004..4507
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4549..4567
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4605..4627)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4727..4746
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4766..4986
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4987..5790
                     /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(5919..6374)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6056..6077)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6268..6287
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6504..6526
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6518..6535
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6519..6535
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6542..6561
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6542..6560
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        6578..7019
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    6578..6695
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     6990..7013
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
ORIGIN
        1 atgtccaaga tcacctcttc tcaagtcaga gaacacgtta aggaattgtt gaagtattct
       61 aacgaaacta agaagagaaa ctttctagaa accgttgaat tgcaagttgg tttgaagaac
      121 tacgacccac aaagagacaa gagattctct ggttccttga agttgcctaa ctgtcctaga
      181 ccaaacatgt ccatctgtat cttcggtgat gctttcgatg ttgacagagc taagtcctgc
      241 ggtgttgacg ctatgtccgt tgatgacttg aagaagttga acaagaacaa gaagttgatt
      301 aagaagttgt ctaagaagta caacgctttc atcgcttccg aagttttgat caagcaagtc
      361 ccacgtttgt tgggtccaca attgtccaag gccggtaagt tcccaacccc agtttctcac
      421 aacgacgatt tgtatggtaa ggttaccgat gtcagatcta ccatcaagtt ccaactaaag
      481 aaggtcttgt gtttggctgt tgctgtcggt aacgtcgaaa tggaagaaga cgttttggtt
      541 aaccaaatct tgatgtctgt caatttcttc gtctctttat tgaagaagaa ctggcaaaac
      601 gttggttcct tggttgttaa gtcttctatg ggtccagctt tcagattgta cgtgagcaag
      661 ggcgaggagc ctagggtgag caagggcgag gaggataaca tggccatcat caaggagttc
      721 atgagattca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      781 gagggcgagg gcagacccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
      841 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
      901 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
      961 aagtgggaga gagtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     1021 tccctgcagg acggcgagtt catctacaag gtgaagttga gaggcaccaa cttcccctcc
     1081 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gagaatgtac
     1141 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     1201 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1261 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccattgtg
     1321 gaacaatatg aaagagctga aggtagacat tctactggtg gtatggatga attgtacaaa
     1381 taagctagca tagattaatt taaacagtat atgtacagtt ttatatatat atatatatat
     1441 atatacatat ataaagaaac ctgtgcgttt tttgtatttt caaatacatt tagttttggc
     1501 gccacttcta taaaaggtct catgatattg ttacagatct gtttagcttg cctcgaggtc
     1561 gactatccat acgatgttcc tgattatgct tagtgtacat gacaccgatt atttaaagct
     1621 gcagcatacg atatatatac atgtgtatat atgtatacct atgaatgtca gtaagtatgt
     1681 atacgaacag tatgatactg aagatgacaa ggtaatgcat cattctatac gtgtcattct
     1741 gaacgaggcg cgcggatccc agcttttgtt ccctttagtg agggttaatc agcttttgtt
     1801 ccctttagtg agggttaatt ccgagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     1861 gaaattgtta tccgctcaca attccacaca acataggagc cggaagcata aagtgtaaag
     1921 cctggggtgc ctaatgagtg aggtaactca cattaattgc gttgcgctca ctgcccgctt
     1981 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     2041 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2101 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2161 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2221 aaaaggccgc gttgctggcg tttttccata ggctcggccc ccctgacgag catcacaaaa
     2281 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgttcc
     2341 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2401 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     2461 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2521 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2581 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2641 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     2701 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2761 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2821 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2881 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2941 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3001 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3061 tagttgcctg actgcccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3121 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3181 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3241 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3301 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3361 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgaaaaaaag
     3421 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3481 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3541 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3601 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3661 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3721 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3781 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3841 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     3901 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3961 ttccgcgcac atttccccga aaagtgccac ctgggtcctt ttcatcacgt gctataaaaa
     4021 taattataat ttaaattttt taatataaat atataaatta aaaatagaaa gtaaaaaaag
     4081 aaattaaaga aaaaatagtt tttgttttcc gaagatgtaa aagactctag ggggatcgcc
     4141 aacaaatact accttttatc ttgctcttcc tgctctcagg tattaatgcc gaattgtttc
     4201 atcttgtctg tgtagaagac cacacacgaa aatcctgtga ttttacattt tacttatcgt
     4261 taatcgaatg tatatctatt taatctgctt ttcttgtcta ataaatatat atgtaaagta
     4321 cgctttttgt tgaaattttt taaacctttg tttatttttt tttcttcatt ccgtaactct
     4381 tctaccttct ttatttactt tctaaaatcc aaatacaaaa cataaaaata aataaacaca
     4441 gagtaaattc ccaaattatt ccatcattaa aagatacgag gcgcgtgtaa gttacaggca
     4501 agcgatccgt cctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     4561 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     4621 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     4681 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     4741 attgtactga gagtgcacca cgcttttcaa ttcaattcat catttttttt ttattctttt
     4801 ttttgatttc ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa
     4861 cgaaggaagg agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca
     4921 tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga
     4981 taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct
     5041 gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt
     5101 cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta
     5161 aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag
     5221 gcattatccg ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt
     5281 aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt
     5341 acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa
     5401 gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc
     5461 ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt
     5521 gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg
     5581 attatgacac ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga
     5641 accgtggatg atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt
     5701 gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca
     5761 tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata
     5821 ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattaccctg cggtgtgaaa
     5881 taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaaacg ttaatatttt
     5941 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     6001 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     6061 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     6121 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     6181 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     6241 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     6301 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     6361 gctacagggc gcgtcgcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     6421 ggtgcgggcc tcttcgctat tacgccagct ggcgaagggg ggatgtgctg caaggcgatt
     6481 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     6541 gtaatacgac tcactatagg gcgaattggc ggccgcacgg attagaagcc gccgagcggg
     6601 tgacagccct ccgaaggaag actctcctcc gtgcgtcctc gtcttcaccg gtcgcgttcc
     6661 tgaaacgcag atgtgcctcg cgccgcactg ctccgaacaa taaagattct acaatactag
     6721 cttttatggt tatgaagagg aaaaattggc agtaacctgg ccccacaaac cttcaaatta
     6781 acgaatcaaa ttaacaacca taggatgata atgcgattag ttttttagcc ttatttctgg
     6841 ggtaattaat cagcgaagcg atgatttttg atctattaac agatatataa atggaaaagc
     6901 tgcataacca ctttaactaa tactttcaac attttcagtt tgtattactt cttattcaaa
     6961 tgtcataaaa gtatcaacaa aaaattgtta atatacctct atactttaac gtcaaggaga
     7021 aggttaatta aggagaaaaa actatagcta gc
//
