LOCUS       sequence-322582-        5542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Backbone plasmid for insertion of the Saccharomyces cerevisiae 
            promoter library and yeGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5542)
  AUTHORS   Renganaath K, Cheung R, Day L, Kosuri S, Kruglyak L, Albert FW
  TITLE     Systematic identification of cis-regulatory variants that cause gene
            expression differences in a yeast cross.
  JOURNAL   Elife. 2020 Nov 12;9. pii: 62669. doi: 10.7554/eLife.62669.
  PUBMED    33179598
REFERENCE   2  (bases 1 to 5542)
  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: "Elife."; 
            date: "2020-11-12"; volume: "9. pii: 62669. doi"; pages: " 
            10.7554/eLife.62669"
COMMENT     Sequence Label: RCP83
FEATURES             Location/Qualifiers
     source          1..5542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             252..968
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1043..1208
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1098..1122)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     misc_feature    1524..2027
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     gene            complement(2107..3463)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      complement(2107..2304)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(2310..3119)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3033..3052
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3120..3463)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      3596..3627
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3628..3730
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(3731..3799,3800..4591)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3949..3968)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             4592..4783
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      4807..4834
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(4846..5433)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4926..4945)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(5455..5484)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 attcctgctg aaccgctctt ccgatctggc cattacggcc aagccggaca ggcagcgaca
       61 gccctgacag acacggaaga ctctcctccg tcctcgtctt caccggtcgc ggaagactct
      121 cctccgtgtg cctacagccg cacggcgcgc cattggcatt atcacataat gaattataca
      181 ttatataaag taatgtgatt tcttcgaaga atatactaaa aaatgagcag gcaagataaa
      241 cgaaggcaaa gatgtctaaa ggtgaagaat tattcactgg tgttgtccca attttggttg
      301 aattagatgg tgatgttaat ggtcacaaat tttctgtctc cggtgaaggt gaaggtgatg
      361 ctacttacgg taaattgacc ttaaaattga tttgtactac tggtaaattg ccagttccat
      421 ggccaacctt agtcactact ttaggttatg gtttgcaatg ttttgctaga tacccagatc
      481 atatgaaaca acatgacttt ttcaagtctg ccatgccaga aggttatgtt caagaaagaa
      541 ctattttttt caaagatgac ggtaactaca agaccagagc tgaagtcaag tttgaaggtg
      601 ataccttagt taatagaatc gaattaaaag gtattgattt taaagaagat ggtaacattt
      661 taggtcacaa attggaatac aactataact ctcacaatgt ttacatcact gctgacaaac
      721 aaaagaatgg tatcaaagct aacttcaaaa ttagacacaa cattgaagat ggtggtgttc
      781 aattagctga ccattatcaa caaaatactc caattggtga tggtccagtc ttgttaccag
      841 acaaccatta cttatcctat caatctgcct tatccaaaga tccaaacgaa aagagagacc
      901 acatggtctt gttagaattt gttactgctg ctggtattac ccatggtatg gatgaattgt
      961 acaaataacc tgcaggctac agcatgggag atacttagat cggaagagcg tcgtgtaggg
     1021 aaagagtgtg gcctaattgg ccgcgaattt cttatgattt atgattttta ttattaaata
     1081 agttataaaa aaaataagtg tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa
     1141 attcttattc ttgagtaact ctttcctgta ggtcaggttg ctttctcagg tatagtatga
     1201 ggtcgctctt attgaccaca ccacagaaga cgggagacac tagcacacaa ctttaccagg
     1261 caaggtattt gacgctagca tgtgtccaat tcagtgtcat ttatgatttt ttgtagtagg
     1321 atataaatat atacagcgct ccaaatagtg cggttgcccc aaaaacacca cggaacctca
     1381 tctgttctcg tactttgttg tgacaaagta gctcactgcc ttattatcac attttcatta
     1441 tgcaacgctt cggaaaatac gatgttgaaa atggatccgg ttccgcgcac atttccccga
     1501 aaagtgccac ctgggtcctt ttcatcacgt gctataaaaa taattataat ttaaattttt
     1561 taatataaat atataaatta aaaatagaaa gtaaaaaaag aaattaaaga aaaaatagtt
     1621 tttgttttcc gaagatgtaa aagactctag ggggatcgcc aacaaatact accttttatc
     1681 ttgctcttcc tgctctcagg tattaatgcc gaattgtttc atcttgtctg tgtagaagac
     1741 cacacacgaa aatcctgtga ttttacattt tacttatcgt taatcgaatg tatatctatt
     1801 taatctgctt ttcttgtcta ataaatatat atgtaaagta cgctttttgt tgaaattttt
     1861 taaacctttg tttatttttt tttcttcatt ccgtaactct tctaccttct ttatttactt
     1921 tctaaaatcc aaatacaaaa cataaaaata aataaacaca gagtaaattc ccaaattatt
     1981 ccatcattaa aagatacgag gcgcgtgtaa gttacaggca agcgatccgg gtaccctcgc
     2041 atgcattaag ggttctcgag agctcgtttt cgacactgga tggcggcgtt agtatcgaat
     2101 cgacagcagt atagcgacca gcattcacat acgattgacg catgatatta ctttctgcgc
     2161 acttaacttc gcatctgggc agatgatgtc gaggcgaaaa aaaatataaa tcacgctaac
     2221 atttgattaa aatagaacaa ctacaatata aaaaaactat acaaatgaca agttcttgaa
     2281 aacaagaatc tttttattgt cagtactgat tagaaaaact catcgagcat caaatgaaac
     2341 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     2401 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     2461 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     2521 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc
     2581 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     2641 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     2701 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     2761 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttgccg
     2821 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     2881 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     2941 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     3001 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     3061 tcagcatcca tgttggaatt taatcgcggc ctcgaaacgt gagtcttttc cttacccatg
     3121 gttgtttatg ttcggatgtg atgtgagaac tgtatcctag caagatttta aaaggaagta
     3181 tatgaaagaa gaacctcagt ggcaaatcct aaccttttat atttctctac aggggcgcgg
     3241 cgtggggaca attcaacgcg tctgtgaggg gagcgtttcc ctgctcgcag gtctgcagcg
     3301 aggagccgta atttttgctt cgcgccgtgc ggccatcaaa atgtatggat gcaaatgatt
     3361 atacatgggg atgtatgggc taaatgtacg ggcgacagtc acatcatgcc cctgagctgc
     3421 gcacgtcaag actgtcaagg agggtattct gggcctccat gtcgctggcc gggtgacccg
     3481 gcggggacga ggcaagctaa acagatctct agacctatct aatctagcgc gacgtcttta
     3541 attaagcggc cgccatcact gaccatttaa atcatacctg acctccatag cagaaagtca
     3601 aaagcctccg accggaggct tttgacttga tcggcacgta agaggttcca actttcacca
     3661 taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt caggagctaa
     3721 ggaagctaaa atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3781 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3841 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3901 tttacgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3961 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4021 gaatgacttg gttgagtact caccagtcac agaaaagcat ctcacggatg gcatgacagt
     4081 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4141 ggcaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4201 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4261 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4321 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggatc
     4381 acttctgcgc tcggccctcc cggctggctg gtttattgct gataaatctg gagccggtga
     4441 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgcatcgt
     4501 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4561 gataggtgcc tcactgatta agcattggta agtgaccaaa caggaaaaaa ccgcccttaa
     4621 catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga
     4681 cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg
     4741 cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgatgagggc ccaaatgtaa
     4801 tcacctggct caccttcggg tgggcctttc tgcgttgctg gcgtttttcc ataggctccg
     4861 cccccctgac gagcatcaca aaaatcgatg ctcaagtcag aggtggcgaa acccgacagg
     4921 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4981 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5041 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5101 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5161 caaccaggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5221 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5281 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttacctcgg aaaaagagtt
     5341 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5401 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatttt ctaccgaaga
     5461 aaggcccacc cgtgaaggtg agccagtgag ttgattgcag tccagttacg ctggagtcaa
     5521 gcgcggccgc ggcggtctcg gc
//
