LOCUS       sequence-317513-        5550 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 5550)
  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 5550)
  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..5550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             691..1407
                     /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      1428..1615
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1483..1507)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1690..3046
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1690..2033
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2034..2843
                     /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(2101..2120)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2849..3046
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3150..3169)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3151..3169)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3256..3273)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3427..4015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3507..3526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4186..4977,4978..5046))
                     /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     4809..4828
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5047..5151)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5219..5237
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5275..5297)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5397..5416
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5497..5515
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5497..5514
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 tcgacacttt gttatgtaga gtttttttag ctacctatat tccaccataa catcaatcat
       61 gcggttgctg gtgtatttac caataatgtt taatgtatat atatatatat atggggccgt
      121 atacttacat atagtagatg tcaagcgtag gcgcttcccc tgccggctgt gagggcgcca
      181 taaccaaggt atctatagac cgccaatcag caaactacct ccgtacattc atgttgcacc
      241 cacacattta tacacccaga ccgcgacaaa ttacccataa ggttgtttgt gacggcgtcg
      301 tacaagagaa cgtgggaact ttttaggctc accaaaaaag aaagaaaaaa tacgagttgc
      361 tgacagaagc ctcaagaaaa aaaaaattct tcttcgacta tgctggaggc agagatgatc
      421 gagccggtag ttaactatat atagctaaat tggttccatc accttctttt ctggtgtcgc
      481 tccttctagt gctatttctg gcttttccta tttttttttt tccatttttc tttctctctt
      541 tctaatatat aaattctctt gcattttcta tttttctctc tatctattct acttgtttat
      601 tcccttcaag gttttttttt aaggagtact tgtttttaga atatacggtc aacgaactat
      661 aattaactaa acctcgagat gttaattaac atgtctaaag gtgaagaatt attcactggt
      721 gttgtcccaa ttttggttga attagatggt gatgttaatg gtcacaaatt ttctgtctcc
      781 ggtgaaggtg aaggtgatgc tacttacggt aaattgacct taaaatttat ttgtactact
      841 ggtaaattgc cagttccatg gccaacctta gtcactactt taacttatgg tgttcaatgt
      901 ttttctagat acccagatca tatgaaacaa catgactttt tcaagtctgc catgccagaa
      961 ggttatgttc aagaaagaac tatttttttc aaagatgacg gtaactacaa gaccagagct
     1021 gaagtcaagt ttgaaggtga taccttagtt aatagaatcg aattaaaagg tattgatttt
     1081 aaagaagatg gtaacatttt aggtcacaaa ttggaataca actataactc tcacaatgtt
     1141 tacatcatgg ctgacaaaca aaagaatggt atcaaagtta acttcaaaat tagacacaac
     1201 attgaagatg gttctgttca attagctgac cattatcaac aaaatactcc aattggtgat
     1261 ggtccagtct tgttaccaga caaccattac ttatccactc aatctgcctt atccaaagat
     1321 ccaaacgaaa agagagacca catggtcttg ttagaatttg ttactgctgc tggtattacc
     1381 catggtattg atgaattgta caaataaggc gcgccacttc taaataagcg aatttcttat
     1441 gatttatgat ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa
     1501 gtgactctta ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca
     1561 ggttgctttc tcaggtatag tatgaggtcg ctcttattga ccacacctct accggcagat
     1621 ccgctaggga taacagggta atatagatct gtttagcttg cctcgtcccc gccgggtcac
     1681 ccggccagcg acatggaggc ccagaatacc ctccttgaca gtcttgacgt gcgcagctca
     1741 ggggcatgat gtgactgtcg cccgtacatt tagcccatac atccccatgt ataatcattt
     1801 gcatccatac attttgatgg ccgcacggcg cgaagcaaaa attacggctc ctcgctgcag
     1861 acctgcgagc agggaaacgc tcccctcaca gacgcgttga attgtcccca cgccgcgccc
     1921 ctgtagagaa atataaaagg ttaggatttg ccactgaggt tcttctttca tatacttcct
     1981 tttaaaatct tgctaggata cagttctcac atcacatccg aacataaaca accatgggta
     2041 aggaaaagac tcacgtttcg aggccgcgat taaattccaa catggatgct gatttatatg
     2101 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg
     2161 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     2221 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     2281 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggcaaaa
     2341 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     2401 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     2461 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     2521 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataagc
     2581 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     2641 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     2701 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     2761 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     2821 tgatgctcga tgagtttttc taatcagtac tgacaataaa aagattcttg ttttcaagaa
     2881 cttgtcattt gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt
     2941 gatttatatt ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa
     3001 agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt cgattcgata
     3061 ctaacgccgc catccagttt aaacgagctc gaattcatcg atgatatcag atccactagt
     3121 ggcctatgcg gccgcggatc tgccggtctc cctatagtga gtcgtattaa tttcgataag
     3181 ccaggttaac ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3241 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3301 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3361 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3421 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3481 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3541 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3601 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3661 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3721 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3781 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3841 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3901 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3961 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4021 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4081 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4141 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4201 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4261 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4321 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4381 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4441 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4501 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4561 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4621 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4681 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4741 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4801 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4861 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4921 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4981 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5041 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5101 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5161 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     5221 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     5281 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     5341 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     5401 atcagagcag attgtactga gagtgcacca tatggacata ttgtcgttag aacgcggcta
     5461 caattaatac ataaccttat gtatcataca catacgattt aggtgacact atagaacgcg
     5521 gccgccagct gaagcttcgt acgctgcagg
//
