LOCUS       sequence-311211-        4897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast codon 
            optimized mTurquoise2 under KanMX selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4897)
  TITLE     Stelling Lab Plasmids
REFERENCE   2  (bases 1 to 4897)
  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: FRP2002
FEATURES             Location/Qualifiers
     source          1..4897
                     /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"
     terminator      829..1016
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(884..908)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1091..2447
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1091..1434
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1435..2244
                     /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(1502..1521)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2250..2447
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2551..2570)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2552..2570)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2657..2674)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2828..3416)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2908..2927)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3587..4378,4379..4447))
                     /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     4210..4229
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4448..4552)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4620..4638
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4676..4698)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4798..4817
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atttaggtga cactatagaa cgcggccgcc agctgaagct tcgtacgctg caggtcgacg
       61 gatcggtgac ggtgctggtt taattaacat ggtttccaag ggtgaagaat tattcactgg
      121 tgttgttcca atcttggttg aattggatgg tgatgttaac ggtcacaagt tttctgtttc
      181 tggtgaaggt gaaggtgatg ctacttatgg taaattgacc ttgaagttca tctgtaccac
      241 aggtaaattg ccagttccat ggccaacttt ggttactact ttgtcttggg gtgttcaatg
      301 ttttgctaga tacccagatc atatgaagca acacgatttc ttcaaatccg ctatgccaga
      361 aggttacgtt caagaaagaa ccatcttctt caaggatgac ggtaactaca aaactagagc
      421 cgaagttaag ttcgaaggtg ataccttggt taacagaatc gaattgaagg gtatcgactt
      481 caaagaagat ggtaacatct tgggtcataa gttggaatac aactacttct ccgataacgt
      541 ttacattacc gccgataagc aaaagaacgg tattaaggct aacttcaaga tcagacacaa
      601 catcgaagat ggtggtgtac aattggctga tcattaccaa caaaacactc caattggtga
      661 tggtccagtt ttgttgccag ataaccatta cttgtctacc caatccaaat tgtctaagga
      721 cccaaacgaa aaaagagatc acatggtctt gttggaattt gttactgctg ctggtattac
      781 cttgggtatg gatgaattat acaagtaagg cgcgccactt ctaaataagc gaatttctta
      841 tgatttatga tttttattat taaataagtt ataaaaaaaa taagtgtata caaattttaa
      901 agtgactctt aggttttaaa acgaaaattc ttattcttga gtaactcttt cctgtaggtc
      961 aggttgcttt ctcaggtata gtatgaggtc gctcttattg accacacctc taccggcaga
     1021 tccgctaggg ataacagggt aatatagatc tgtttagctt gcctcgtccc cgccgggtca
     1081 cccggccagc gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc
     1141 aggggcatga tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt
     1201 tgcatccata cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgca
     1261 gacctgcgag cagggaaacg ctcccctcac agacgcgttg aattgtcccc acgccgcgcc
     1321 cctgtagaga aatataaaag gttaggattt gccactgagg ttcttctttc atatacttcc
     1381 ttttaaaatc ttgctaggat acagttctca catcacatcc gaacataaac aaccatgggt
     1441 aaggaaaaga ctcacgtttc gaggccgcga ttaaattcca acatggatgc tgatttatat
     1501 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat
     1561 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     1621 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     1681 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggcaaa
     1741 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     1801 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     1861 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     1921 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataag
     1981 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     2041 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     2101 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     2161 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     2221 ttgatgctcg atgagttttt ctaatcagta ctgacaataa aaagattctt gttttcaaga
     2281 acttgtcatt tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg
     2341 tgatttatat tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga
     2401 aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat
     2461 actaacgccg ccatccagtt taaacgagct cgaattcatc gatgatatca gatccactag
     2521 tggcctatgc ggccgcggat ctgccggtct ccctatagtg agtcgtatta atttcgataa
     2581 gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     2641 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2701 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2761 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2821 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2881 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2941 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3001 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3061 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3121 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3181 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3241 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3301 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3361 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3421 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3481 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3541 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3601 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3661 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3721 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3781 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3841 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3901 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3961 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4021 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4081 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4141 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4201 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4261 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4321 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4381 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4441 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4501 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4561 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     4621 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     4681 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     4741 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     4801 catcagagca gattgtactg agagtgcacc atatggacat attgtcgtta gaacgcggct
     4861 acaattaata cataacctta tgtatcatac acatacg
//
