LOCUS       sequence-318111-        5554 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses eGFP under control of the yeast TEF2 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5554)
  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 5554)
  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: pBAC690-TEF
FEATURES             Location/Qualifiers
     source          1..5554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             690..1406
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGIDELYK"
     terminator      1427..1614
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1482..1506)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1689..3045
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1689..2032
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2033..2842
                     /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     complement(2100..2119)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2848..3045
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3149..3168)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3150..3168)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3255..3272)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3426..4014)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3506..3525)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4185..4976,4977..5045))
                     /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     4808..4827
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5046..5150)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5218..5236
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5274..5296)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5396..5415
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5496..5514
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5496..5513
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 actttgttat gtagagtttt tttagctacc tatattccac cataacatca atcatgcggt
       61 tgctggtgta tttaccaata atgtttaatg tatatatata tatatatata tggggccgta
      121 tacttacata tagtagatgt caagcgtagg cgcttcccct gccggctgtg agggcgccat
      181 aaccaaggta tctatagacc gccaatcagc aaactacctc cgtacattca tgttgcaccc
      241 acacatttat acacccagac cgcgacaaat tacccataag gttgtttgtg acggcgtcgt
      301 acaagagaac gtgggaactt tttaggctca ccaaaaaaga aagaaaaaat acgagttgct
      361 gacagaagcc tcaagaaaaa aaaaattctt cttcgactat gctggaggca gagatgatcg
      421 agccggtagt taactatata tagctaaatt ggttccatca ccttcttttc tggtgtcgct
      481 ccttctagtg ctatttctgg cttttcctat tttttttttt ccatttttct ttctctcttt
      541 ctaatatata aattctcttg cattttctat ttttctctct atctattcta cttgtttatt
      601 cccttcaagg ttttttttta aggagtactt gtttttagaa tatacggtca acgaactata
      661 attaactaaa cctcgagatg ttaattaaca tgtctaaagg tgaagaatta ttcactggtg
      721 ttgtcccaat tttggttgaa ttagatggtg atgttaatgg tcacaaattt tctgtctccg
      781 gtgaaggtga aggtgatgct acttacggta aattgacctt aaaatttatt tgtactactg
      841 gtaaattgcc agttccatgg ccaaccttag tcactacttt aacttatggt gttcaatgtt
      901 tttctagata cccagatcat atgaaacaac atgacttttt caagtctgcc atgccagaag
      961 gttatgttca agaaagaact atttttttca aagatgacgg taactacaag accagagctg
     1021 aagtcaagtt tgaaggtgat accttagtta atagaatcga attaaaaggt attgatttta
     1081 aagaagatgg taacatttta ggtcacaaat tggaatacaa ctataactct cacaatgttt
     1141 acatcatggc tgacaaacaa aagaatggta tcaaagttaa cttcaaaatt agacacaaca
     1201 ttgaagatgg ttctgttcaa ttagctgacc attatcaaca aaatactcca attggtgatg
     1261 gtccagtctt gttaccagac aaccattact tatccactca atctgcctta tccaaagatc
     1321 caaacgaaaa gagagaccac atggtcttgt tagaatttgt tactgctgct ggtattaccc
     1381 atggtattga tgaattgtac aaataaggcg cgccacttct aaataagcga atttcttatg
     1441 atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca aattttaaag
     1501 tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc tgtaggtcag
     1561 gttgctttct caggtatagt atgaggtcgc tcttattgac cacacctcta ccggcagatc
     1621 cgctagggat aacagggtaa tatagatctg tttagcttgc ctcgtccccg ccgggtcacc
     1681 cggccagcga catggaggcc cagaataccc tccttgacag tcttgacgtg cgcagctcag
     1741 gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta taatcatttg
     1801 catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga
     1861 cctgcgagca gggaaacgct cccctcacag acgcgttgaa ttgtccccac gccgcgcccc
     1921 tgtagagaaa tataaaaggt taggatttgc cactgaggtt cttctttcat atacttcctt
     1981 ttaaaatctt gctaggatac agttctcaca tcacatccga acataaacaa ccatgggtaa
     2041 ggaaaagact cacgtttcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     2101 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg
     2161 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     2221 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     2281 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggcaaaac
     2341 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     2401 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     2461 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     2521 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataagct
     2581 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     2641 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     2701 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     2761 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     2821 gatgctcgat gagtttttct aatcagtact gacaataaaa agattcttgt tttcaagaac
     2881 ttgtcatttg tatagttttt ttatattgta gttgttctat tttaatcaaa tgttagcgtg
     2941 atttatattt tttttcgcct cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa
     3001 gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta tactgctgtc gattcgatac
     3061 taacgccgcc atccagttta aacgagctcg aattcatcga tgatatcaga tccactagtg
     3121 gcctatgcgg ccgcggatct gccggtctcc ctatagtgag tcgtattaat ttcgataagc
     3181 caggttaacc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3241 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3301 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3361 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3421 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3481 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3541 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3601 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3661 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3721 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3781 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3841 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     3901 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3961 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4021 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4081 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4141 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4201 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4261 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4321 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4381 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4441 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4501 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4561 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4621 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4681 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4741 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4801 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4861 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4921 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4981 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5041 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5101 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5161 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     5221 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     5281 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     5341 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     5401 tcagagcaga ttgtactgag agtgcaccat atggacatat tgtcgttaga acgcggctac
     5461 aattaataca taaccttatg tatcatacac atacgattta ggtgacacta tagaacgcgg
     5521 ccgccagctg aagcttcgta cgctgcaggt cgac
//
