LOCUS       sequence-489756-        4908 bp    DNA     circular SYN 20-OCT-2025
DEFINITION  Yeast tagging vector to create a gene fusion with yeast optimized 
            mNeongreen with Kan selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4908)
  TITLE     Teusink Lab yeast expression plasmids
REFERENCE   2  (bases 1 to 4908)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 20, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pFA6a-link-ymNeongreen-Kan
FEATURES             Location/Qualifiers
     source          1..4908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     125..143
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(181..203)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     303..322
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        403..421
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     403..420
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      746..755
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      1242..1429
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1297..1321)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1504..2860
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1504..1847
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1848..2657
                     /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(1915..1934)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2663..2860
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2964..2983)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2965..2983)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3070..3087)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3241..3829)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3321..3340)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4000..4791,4792..4860))
                     /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     4623..4642
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4861..4908,1..57))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
       61 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
      121 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
      181 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
      241 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
      301 tgcggcatca gagcagattg tactgagagt gcaccatatg gacatattgt cgttagaacg
      361 cggctacaat taatacataa ccttatgtat catacacata cgatttaggt gacactatag
      421 aacgcggccg ccagctgaag cttcgtacgc tgcaggtcga cggatcggtg acggtgctgg
      481 tttaattaac atggtctcta agggtgaaga agataacatg gcttctttgc cagccaccca
      541 cgagttgcac atcttcggtt ccattaacgg tgtcgacttc gatatggtcg gtcaaggtac
      601 cggtaaccca aacgacggtt acgaagaatt gaacttgaag tctaccaagg gcgacttgca
      661 gttctcccct tggattttgg ttccacacat cggttacggt ttccaccaat acttgccata
      721 cccagacggt atgtccccat tccaagccgc catggttgac ggttctggtt accaagtcca
      781 cagaaccatg caattcgaag acggtgcttc cttgaccgtg aactacagat acacctacga
      841 aggttctcac atcaagggcg aagcccaagt taagggtacc ggtttccctg ctgacggtcc
      901 agtcatgact aactccttga ccgctgctga ctggtgtcgt tccaagaaga cctacccaaa
      961 cgataagacc atcatttcca ctttcaagtg gtcctacacc accggtaacg gcaagagata
     1021 ccgctctacc gctcgtacca cctacacctt cgccaagcca atggctgcca actacttgaa
     1081 gaaccagcct atgtacgtct tccgtaagac cgagctgaag cattccaaga ctgaattgaa
     1141 cttcaaggaa tggcaaaagg ctttcaccga cgtcatgggt atggacgaat tgtacaagta
     1201 agcggccgcc tcgaggtcga cggcgcgcca cttctaaata agcgaatttc ttatgattta
     1261 tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact
     1321 cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc
     1381 tttctcaggt atagtatgag gtcgctctta ttgaccacac ctctaccggc agatccgcta
     1441 gggataacag ggtaatatag atctgtttag cttgcctcgt ccccgccggg tcacccggcc
     1501 agcgacatgg aggcccagaa taccctcctt gacagtcttg acgtgcgcag ctcaggggca
     1561 tgatgtgact gtcgcccgta catttagccc atacatcccc atgtataatc atttgcatcc
     1621 atacattttg atggccgcac ggcgcgaagc aaaaattacg gctcctcgct gcagacctgc
     1681 gagcagggaa acgctcccct cacagacgcg ttgaattgtc cccacgccgc gcccctgtag
     1741 agaaatataa aaggttagga tttgccactg aggttcttct ttcatatact tccttttaaa
     1801 atcttgctag gatacagttc tcacatcaca tccgaacata aacaaccatg ggtaaggaaa
     1861 agactcacgt ttcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     1921 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc
     1981 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     2041 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     2101 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccggc aaaacagcat
     2161 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     2221 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     2281 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     2341 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aagcttttgc
     2401 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     2461 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     2521 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     2581 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     2641 tcgatgagtt tttctaatca gtactgacaa taaaaagatt cttgttttca agaacttgtc
     2701 atttgtatag tttttttata ttgtagttgt tctattttaa tcaaatgtta gcgtgattta
     2761 tatttttttt cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat
     2821 atcatgcgtc aatcgtatgt gaatgctggt cgctatactg ctgtcgattc gatactaacg
     2881 ccgccatcca gtttaaacga gctcgaattc atcgatgata tcagatccac tagtggccta
     2941 tgcggccgcg gatctgccgg tctccctata gtgagtcgta ttaatttcga taagccaggt
     3001 taacctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3061 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3121 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3181 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3241 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3301 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3361 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3421 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3481 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3541 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3601 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3661 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3721 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3781 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3841 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3901 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3961 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4021 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4081 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4141 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4201 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4261 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4321 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4381 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4441 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4501 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4561 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4621 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4681 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4741 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4801 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4861 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtct
//
