LOCUS       sequence-390052-        7647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TEF1 promoter controlled expression plasmid with G418 resistance for
            expressing ymTurquoise2 in budding yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7647)
  TITLE     Expression plasmids with KanMX/G418 marker
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7647)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCEV-G2-Km ymTurquoise2
FEATURES             Location/Qualifiers
     source          1..7647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(2..167)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     88..112
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(315..338)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        1354..1760
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     1780..1799
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1780..1798
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2541..2730
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2802..2819)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2973..3561)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3053..3072)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3732..4523,4524..4592))
                     /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     4355..4374
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4593..4697)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4724..6066
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5687..5734)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     protein_bind    6086..6119
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            6174..7530
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        6174..6517
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             6518..7327
                     /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(6585..6604)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      7333..7530
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    7593..7626
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 ggagcgacct catgctatac ctgagaaagc aacctgacct acaggaaaga gttactcaag
       61 aataagaatt ttcgttttaa aacctaagag tcactttaaa atttgtatac acttattttt
      121 tttataactt atttaataat aaaaatcata aatcataaga aattcgctta tttagaagtg
      181 tcaacaacgt atctaccaac gatttgaccc ttttccatct tttcgtaaat ttctggcaag
      241 gtagacaagc cgacaacctt gattggagac ttgaccaaac ctctggcgaa gaattgttaa
      301 ttaagagctc agatcttatc gtcgtcatcc ttgtaatcca tcgatactag tgcggccgct
      361 tgttttatat ttgttgtaaa aagtagataa ttacttcctt gatgatctgt aaaaaagaga
      421 aaaagaaagc atctaagaac ttgaaaaact acgaattaga aaagaccaaa tatgtatttc
      481 ttgcattgac caatttatgc aagtttatat atatgtaaat gtaagtttca cgaggttcta
      541 ctaaactaaa ccaccccctt ggttagaaga aaagagtgtg tgagaacagg ctgttgttgt
      601 cacacgattc ggacaattct gtttgaaaga gagagagtaa cagtacgatc gaacgaactt
      661 tgctctggag atcacagtgg gcatcatagc atgtggtact aaaccctttc ccgccattcc
      721 agaaccttcg attgcttgtt acaaaacctg tgagccgtcg ctaggacctt gttgtgtgac
      781 gaaattggaa gctgcaatca ataggaagac aggaagtcga gcgtgtctgg gttttttcag
      841 ttttgttctt tttgcaaaca aatcacgagc gacggtaatt tctttctcga taagaggcca
      901 cgtgctttat gagggtaaca tcaattcaag aaggagggaa acacttcctt tttctggccc
      961 tgataatagt atgagggtga agccaaaata aaggattcgc gcccaaatcg gcatctttaa
     1021 atgcaggtat gcgatagttc ctcactcttt ccttactcac gagtaattct tgcaaatgcc
     1081 tattatgcag atgttataat atctgtgcgt cttgagttga agtcaggaat ctaaaataaa
     1141 aattaaggtt aataaaaaga ggaaagaaaa aaaaattaat cgatttacag aaacttgcac
     1201 actaaaaata cacaactaaa agcaattaca gtatgggaag tcatcgacgt tatctctact
     1261 atagtatatt atcatttcta ttattatcct gctcagtggt acttgcaaaa caagataaga
     1321 ccccattctt tgaaggtact tccaggccgg ccgcacacac catagcttca aaatgtttct
     1381 actccttttt tactcttcca gattttctcg gactccgcgc atcgccgtac cacttcaaaa
     1441 cacccaagca cagcatacta aatttcccct ctttcttcct ctagggtgtc gttaattacc
     1501 cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt ttctttttct tcgtcgaaaa
     1561 aggcaataaa aatttttatc acgtttcttt ttcttgaaaa tttttttttt tgattttttt
     1621 ctctttcgat gacctcccat tgatatttaa gttaataaac ggtcttcaat ttctcaagtt
     1681 tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgctcat tagaaagaaa
     1741 gcatagcaat ctaatctaag ttttaattac aaggatccgt aatacgactc actatagggc
     1801 ccgggatggt tagtaaaggt gaagaattgt tcactggtgt agtgccaatt ttagtagaat
     1861 tagatggtga cgttaacggt cataagttct ctgtaagcgg tgaaggtgaa ggtgacgcaa
     1921 catacggtaa attaactttg aaattcattt gtactacagg taaattacca gttccatggc
     1981 caactttggt tactactttg tcttggggtg ttcaatgttt cgctagatac ccagatcata
     2041 tgaagcaaca tgatttcttt aagtctgcta tgccagaagg ttacgttcaa gaaagaacta
     2101 tcttcttcaa ggatgatggt aactataaga ctagagctga agttaagttc gaaggagata
     2161 ctttagttaa tagaattgaa ttgaagggta ttgattttaa ggaagatggt aatattttgg
     2221 gtcataagtt ggaatataat tacttctctg ataacgttta cattactgct gataagcaaa
     2281 agaacggtat taaggcgaac ttcaagatta gacataacat tgaagatggt ggtgttcaat
     2341 tagctgatca ttaccaacaa aacactccaa ttggtgacgg tccagttttg ttgccagata
     2401 accattactt gtctactcaa tctaagttgt ctaaggatcc aaacgaaaag agagatcata
     2461 tggttttgtt ggaattcgtt actgctgctg ggatcacttt aggtatggat gaattgtata
     2521 aataaccgcg gctagctaag atccgctcta accgaaaagg aaggagttag acaacctgaa
     2581 gtctaggtcc ctatttattt ttttatagtt atgttagtat taagaacgtt atttatattt
     2641 caaatttttc ttttttttct gtacagacgc gtgtacgcat gtaacattat actgaaaacc
     2701 ttgcttgaga aggttttggg acgctcgaag atccagctgc attaatgaat cggccaacgc
     2761 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2821 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2881 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2941 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3001 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3061 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3121 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3181 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3241 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3301 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3361 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3421 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3481 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3541 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3601 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3661 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3721 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3781 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3841 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3901 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3961 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4021 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4081 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4141 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4201 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4261 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4321 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4381 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4441 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4501 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4561 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4621 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4681 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaacgaag catctgtgct
     4741 tcattttgta gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag
     4801 ctgcattttt acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg
     4861 tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat
     4921 ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga
     4981 atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa
     5041 gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt
     5101 tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc
     5161 ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca
     5221 ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt
     5281 tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt
     5341 cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg
     5401 attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata
     5461 aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg
     5521 taggttatat agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt
     5581 tgtggaagcg gtattcgcaa tattttagta gctcgttaca gtccggtgcg tttttggttt
     5641 tttgaaagtg cgtcttcaga gcgcttttgg ttttcaaaag cgctctgaag ttcctatact
     5701 ttctagagaa taggaacttc ggaataggaa cttcaaagcg tttccgaaaa cgagcgcttc
     5761 cgaaaatgca acgcgagctg cgcacataca gctcactgtt cacgtcgcac ctatatctgc
     5821 gtgttgcctg tatatatata tacatgagaa gaacggcata gtgcgtgttt atgcttaaat
     5881 gcgtacttat atgcgtctat ttatgtagga tgaaaggtag tctagtacct cctgtgatat
     5941 tatcccattc catgcggggt atcgtatgct tccttcagca ctacccttta gctgttctat
     6001 atgctgccac tcctcaattg gattagtctc atccttcaat gctatcattt cctttgatat
     6061 tggatcatgg tagacaaccc ttaatataac ttcgtataat gtatgctata cgaagttatt
     6121 aggtctagag atctgtttag cttgcctcgt ccccgccggg tcacccggcc agcgacatgg
     6181 aggcccagaa taccctcctt gacagtcttg acgtgcgcag ctcaggggca tgatgtgact
     6241 gtcgcccgta catttagccc atacatcccc atgtataatc atttgcatcc atacattttg
     6301 atggccgcac ggcgcgaagc aaaaattacg gctcctcgct gcagacctgc gagcagggaa
     6361 acgctcccct cacagacgcg ttgaattgtc cccacgccgc gcccctgtag agaaatataa
     6421 aaggttagga tttgccactg aggttcttct ttcatatact tccttttaaa atcttgctag
     6481 gatacagttc tcacatcaca tccgaacata aacaaccatg ggtaaggaaa agactcacgt
     6541 ttcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     6601 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc
     6661 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     6721 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     6781 tcctgatgat gcatggttac tcaccactgc gatccccggc aaaacagcat tccaggtatt
     6841 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     6901 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     6961 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     7021 taatggctgg cctgttgaac aagtctggaa agaaatgcat aagcttttgc cattctcacc
     7081 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     7141 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     7201 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     7261 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     7321 tttctaatca gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag
     7381 tttttttata ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt
     7441 cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc
     7501 aatcgtatgt gaatgctggt cgctatactg ctgtcgattc gatactaacg ccgccatcca
     7561 gtgtcgaaaa cgagctctcg agaaccctta atataacttc gtataatgta tgctatacga
     7621 agttattagg tgatatcaga tccacta
//
