LOCUS       sequence-311201-        4910 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with sfGFP under KanMX 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4910)
  TITLE     Stelling Lab Plasmids
REFERENCE   2  (bases 1 to 4910)
  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"
COMMENT     Sequence Label: FRP272
FEATURES             Location/Qualifiers
     source          1..4910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1..18
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             join(89..91,92..805)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             89..802
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(119..147)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     742..762
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      842..1029
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(897..921)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1104..2460
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1104..1447
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1448..2257
                     /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(1515..1534)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2263..2460
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2564..2583)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2565..2583)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2670..2687)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2841..3429)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2921..2940)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3600..4391,4392..4460))
                     /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     4223..4242
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4461..4565)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4633..4651
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4689..4711)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4811..4830
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atttaggtga cactatagaa cgcggccgcc agctgaagct tcgtacgctg caggtcgacg
       61 gatcggtgac ggtgctggtt taattaacat gagtaaagga gaagaacttt tcactggagt
      121 tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtccgtgg
      181 agagggtgaa ggtgatgcta caaacggaaa actcaccctt aaatttattt gcactactgg
      241 aaaactacct gttccgtggc caacacttgt cactactctg acctatggtg ttcaatgctt
      301 ttcccgttat ccggatcaca tgaaacggca tgactttttc aagagtgcca tgcccgaagg
      361 ttatgtacag gaacgcacta tatctttcaa agatgacggg acctacaaga cgcgtgctga
      421 agtcaagttt gaaggtgata cccttgttaa tcgtatcgag ttaaagggta ttgattttaa
      481 agaagatgga aacattcttg gacacaaact cgagtacaac tttaactcac acaatgtata
      541 catcacggca gacaaacaaa agaatggaat caaagctaac ttcaaaattc gccacaacgt
      601 tgaagatggt tccgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg
      661 ccctgtcctt ttaccagaca accattacct gtcgacacaa tctgtccttt cgaaagatcc
      721 caacgaaaag cgtgaccaca tggtccttct tgagtttgta actgctgctg ggattacaca
      781 tggcatggat gagctctaca aataataagc ttggctgttt tggcgcgcca cttctaaata
      841 agcgaatttc ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt
      901 atacaaattt taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc
      961 tttcctgtag gtcaggttgc tttctcaggt atagtatgag gtcgctctta ttgaccacac
     1021 ctctaccggc agatccgcta gggataacag ggtaatatag atctgtttag cttgcctcgt
     1081 ccccgccggg tcacccggcc agcgacatgg aggcccagaa taccctcctt gacagtcttg
     1141 acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc
     1201 atgtataatc atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg
     1261 gctcctcgct gcagacctgc gagcagggaa acgctcccct cacagacgcg ttgaattgtc
     1321 cccacgccgc gcccctgtag agaaatataa aaggttagga tttgccactg aggttcttct
     1381 ttcatatact tccttttaaa atcttgctag gatacagttc tcacatcaca tccgaacata
     1441 aacaaccatg ggtaaggaaa agactcacgt ttcgaggccg cgattaaatt ccaacatgga
     1501 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     1561 ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     1621 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     1681 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     1741 gatccccggc aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     1801 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     1861 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     1921 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     1981 agaaatgcat aagcttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     2041 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     2101 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     2161 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     2221 attgcagttt catttgatgc tcgatgagtt tttctaatca gtactgacaa taaaaagatt
     2281 cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt tctattttaa
     2341 tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc agatgcgaag
     2401 ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt cgctatactg
     2461 ctgtcgattc gatactaacg ccgccatcca gtttaaacga gctcgaattc atcgatgata
     2521 tcagatccac tagtggccta tgcggccgcg gatctgccgg tctccctata gtgagtcgta
     2581 ttaatttcga taagccaggt taacctgcat taatgaatcg gccaacgcgc ggggagaggc
     2641 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2701 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2761 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2821 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2881 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2941 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3001 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3061 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3121 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3181 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3241 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3301 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3361 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3421 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3481 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3541 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3601 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3661 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3721 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3781 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3841 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3901 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3961 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4021 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4081 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4141 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4201 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4261 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4321 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4381 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4441 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4501 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4561 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     4621 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     4681 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     4741 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     4801 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgga catattgtcg
     4861 ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg
//
