LOCUS       sequence-383892-        7626 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TEF1 promoter controlled expression plasmid with G418 resistance for
            expressing ymScarletI in budding yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7626)
  TITLE     Expression plasmids with KanMX/G418 marker
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7626)
  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            "Unpublished"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 ymScarletI
FEATURES             Location/Qualifiers
     source          1..7626
                     /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      2520..2709
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2781..2798)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2952..3540)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3032..3051)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3711..4502,4503..4571))
                     /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     4334..4353
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4572..4676)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4703..6045
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5666..5713)
                     /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    6065..6098
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            6153..7509
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        6153..6496
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             6497..7306
                     /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(6564..6583)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      7312..7509
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    7572..7605
                     /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 gaagctgtta taaaagaatt catgaggttt aaagttcata
     1861 tggaaggttc aatgaatggt catgaatttg aaattgaagg tgaaggtgaa ggtagaccat
     1921 atgaaggtac acaaactgct aaattgaaag ttactaaagg tggtccatta ccattttctt
     1981 gggatatttt gtctccacaa ttcatgtatg gttctagagc ttttattaag catccagctg
     2041 atattccaga ttattataaa caatcttttc ctgaaggttt taaatgggaa agagttatga
     2101 atttcgaaga tggtggtgct gttactgtta ctcaagatac ttctttggaa gatggtactt
     2161 taatctataa agttaaattg agaggtacta attttccacc agatggtcca gttatgcaaa
     2221 agaaaactat gggttgggaa gcatctactg aaagattgta tccagaagat ggtgttttga
     2281 aaggagacat taagatggct ttgagattga aagatggtgg tcggtacctg gccgacttca
     2341 agaccaccta taaagctaaa aaaccagttc aaatgccagg tgcatataat gttgatagaa
     2401 agttagatat aacgtcgcat aacgaggact atactgtagt tgaacaatat gaacgtagtg
     2461 aaggtagaca tagtaccgga ggaatggatg aattgtataa ataaccgcgg ctagctaaga
     2521 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt
     2581 tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     2641 tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga
     2701 cgctcgaaga tccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     2761 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2821 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2881 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2941 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3001 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3061 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3121 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3181 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3241 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3301 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3361 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3421 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3481 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3541 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3601 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3661 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3721 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3781 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3841 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3901 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3961 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4021 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4081 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4141 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4201 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4261 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4321 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4381 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4441 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4501 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4561 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4621 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4681 tttccccgaa aagtgccacc tgaacgaagc atctgtgctt cattttgtag aacaaaaatg
     4741 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     4801 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
     4861 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     4921 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     4981 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     5041 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     5101 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     5161 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     5221 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     5281 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     5341 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     5401 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     5461 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     5521 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     5581 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcttcagag
     5641 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagagaat aggaacttcg
     5701 gaataggaac ttcaaagcgt ttccgaaaac gagcgcttcc gaaaatgcaa cgcgagctgc
     5761 gcacatacag ctcactgttc acgtcgcacc tatatctgcg tgttgcctgt atatatatat
     5821 acatgagaag aacggcatag tgcgtgttta tgcttaaatg cgtacttata tgcgtctatt
     5881 tatgtaggat gaaaggtagt ctagtacctc ctgtgatatt atcccattcc atgcggggta
     5941 tcgtatgctt ccttcagcac taccctttag ctgttctata tgctgccact cctcaattgg
     6001 attagtctca tccttcaatg ctatcatttc ctttgatatt ggatcatggt agacaaccct
     6061 taatataact tcgtataatg tatgctatac gaagttatta ggtctagaga tctgtttagc
     6121 ttgcctcgtc cccgccgggt cacccggcca gcgacatgga ggcccagaat accctccttg
     6181 acagtcttga cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac atttagccca
     6241 tacatcccca tgtataatca tttgcatcca tacattttga tggccgcacg gcgcgaagca
     6301 aaaattacgg ctcctcgctg cagacctgcg agcagggaaa cgctcccctc acagacgcgt
     6361 tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat ttgccactga
     6421 ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct cacatcacat
     6481 ccgaacataa acaaccatgg gtaaggaaaa gactcacgtt tcgaggccgc gattaaattc
     6541 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     6601 tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     6661 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     6721 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     6781 caccactgcg atccccggca aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     6841 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     6901 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     6961 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     7021 agtctggaaa gaaatgcata agcttttgcc attctcaccg gattcagtcg tcactcatgg
     7081 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     7141 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     7201 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     7261 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag tactgacaat
     7321 aaaaagattc ttgttttcaa gaacttgtca tttgtatagt ttttttatat tgtagttgtt
     7381 ctattttaat caaatgttag cgtgatttat attttttttc gcctcgacat catctgccca
     7441 gatgcgaagt taagtgcgca gaaagtaata tcatgcgtca atcgtatgtg aatgctggtc
     7501 gctatactgc tgtcgattcg atactaacgc cgccatccag tgtcgaaaac gagctctcga
     7561 gaacccttaa tataacttcg tataatgtat gctatacgaa gttattaggt gatatcagat
     7621 ccacta
//
