LOCUS       sequence-317510-        7022 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 7022)
  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 7022)
  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..7022
                     /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(1772..1790)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1811..1827)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1811..1827)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1824..1846)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1835..1851
                     /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(1859..1865,1866..1883,1884..1889))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1904..1925
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2042..2059)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2213..2801)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2293..2312)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2972..3763,3764..3832))
                     /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     3595..3614
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3833..3937)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3974..4477
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4518..4536
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4574..4596)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4696..4715
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4739..4954
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4955..5758
                     /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(5889..6344)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6026..6047)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6238..6257
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6474..6496
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6488..6505
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6489..6505
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6512..6531
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6512..6530
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        6548..6989
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    6548..6665
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     6960..6983
                     /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 agggttaatt tcgagcttgg
     1801 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     1861 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     1921 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     1981 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2041 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2101 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2161 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2221 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2281 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2341 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2401 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2461 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2521 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2581 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2641 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2701 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2761 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2821 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2881 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2941 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3001 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3061 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3121 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3181 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3241 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3301 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3361 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3421 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3481 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3541 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3601 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3661 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3721 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3781 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     3841 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3901 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     3961 ctgggtcctt ttcatcacgt gctataaaaa taattataat ttaaattttt taatataaat
     4021 atataaatta aaaatagaaa gtaaaaaaag aaattaaaga aaaaatagtt tttgttttcc
     4081 gaagatgtaa aagactctag ggggatcgcc aacaaatact accttttatc ttgctcttcc
     4141 tgctctcagg tattaatgcc gaattgtttc atcttgtctg tgtagaagac cacacacgaa
     4201 aatcctgtga ttttacattt tacttatcgt taatcgaatg tatatctatt taatctgctt
     4261 ttcttgtcta ataaatatat atgtaaagta cgctttttgt tgaaattttt taaacctttg
     4321 tttatttttt tttcttcatt ccgtaactct tctaccttct ttatttactt tctaaaatcc
     4381 aaatacaaaa cataaaaata aataaacaca gagtaaattc ccaaattatt ccatcattaa
     4441 aagatacgag gcgcgtgtaa gttacaggca agcgatccgt ctaagaaacc attattatca
     4501 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     4561 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     4621 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     4681 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat accacctttt
     4741 caattcatca tttttttttt attctttttt ttgatttcgg tttccttgaa atttttttga
     4801 ttcggtaatc tccgaacaga aggaagaacg aaggaaggag cacagactta gattggtata
     4861 tatacgcata tgtagtgttg aagaaacatg aaattgccca gtattcttaa cccaactgca
     4921 cagaacaaaa acctgcagga aacgaagata aatcatgtcg aaagctacat ataaggaacg
     4981 tgctgctact catcctagtc ctgttgctgc caagctattt aatatcatgc acgaaaagca
     5041 aacaaacttg tgtgcttcat tggatgttcg taccaccaag gaattactgg agttagttga
     5101 agcattaggt cccaaaattt gtttactaaa aacacatgtg gatatcttga ctgatttttc
     5161 catggagggc acagttaagc cgctaaaggc attatccgcc aagtacaatt ttttactctt
     5221 cgaagacaga aaatttgctg acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt
     5281 atacagaata gcagaatggg cagacattac gaatgcacac ggtgtggtgg gcccaggtat
     5341 tgttagcggt ttgaagcagg cggcagaaga agtaacaaag gaacctagag gccttttgat
     5401 gttagcagaa ttgtcatgca agggctccct atctactgga gaatatacta agggtactgt
     5461 tgacattgcg aagagcgaca aagattttgt tatcggcttt attgctcaaa gagacatggg
     5521 tggaagagat gaaggttacg attggttgat tatgacaccc ggtgtgggtt tagatgacaa
     5581 gggagacgca ttgggtcaac agtatagaac cgtggatgat gtggtctcta caggatctga
     5641 cattattatt gttggaagag gactatttgc aaagggaagg gatgctaagg tagagggtga
     5701 acgttacaga aaagcaggct gggaagcata tttgagaaga tgcggccagc aaaactaaaa
     5761 aactgtatta taagtaaatg catgtatact aaactcacaa attagagctt caatttaatt
     5821 atatcagtta ttaccctatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     5881 gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     5941 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     6001 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     6061 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     6121 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     6181 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     6241 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     6301 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcgcgcc attcgccatt
     6361 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     6421 ggcgaagggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     6481 acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactatagg gcgaattggc
     6541 ggccgcacgg attagaagcc gccgagcggg cgacagccct ccgacggaag actctcctcc
     6601 gtgcgtcctc gtcttcaccg gtcgcgttcc tgaaacgcag atgtgcctcg cgccgcactg
     6661 ctccgaacaa taaagattct acaatactag cttttatggt tatgaagagg aaaaattggc
     6721 agtaacctgg ccccacaaac cttcaaatta acgaatcaaa ttaacaacca taggatgata
     6781 atgcgattag ttttttagcc ttatttctgg ggtaattaat cagcgaagcg atgatttttg
     6841 atctattaac agatatataa atggaaaagc tgcataacca ctttaactaa tactttcaac
     6901 attttcagtt tgtattactt cttattcaaa tgtcataaaa gtatcaacaa aaaattgtta
     6961 atatacctct atactttaac gtcaaggaga aggttaatta aggagaaaaa actatagcta
     7021 gc
//
