LOCUS       sequence-311212-        4831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast codon 
            optimized mKOkappa under KanMX selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4831)
  TITLE     Stelling Lab Plasmids
REFERENCE   2  (bases 1 to 4831)
  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: FRP2008
FEATURES             Location/Qualifiers
     source          1..4831
                     /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      763..950
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(818..842)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1025..2381
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1025..1368
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1369..2178
                     /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(1436..1455)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2184..2381
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2485..2504)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2486..2504)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2591..2608)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2762..3350)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2842..2861)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3521..4312,4313..4381))
                     /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     4144..4163
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4382..4486)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4554..4572
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4610..4632)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4732..4751
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atttaggtga cactatagaa cgcggccgcc agctgaagct tcgtacgctg caggtcgacg
       61 gatcggtgac ggtgctggtt taattaacat gtccgtcatc aagccagaaa tgaagatgag
      121 atattacatg gacggttccg ttaacggtca tgaattcact attgaaggtg aaggtactgg
      181 tagaccatat gaaggtcatc aagaaatgac tttgagagtt actatggctg aaggtggtcc
      241 aatgccattt gcttttgatt tggtttctca cgttttctgc tacggtcata gagtttttac
      301 taagtaccca gaagaaatcc cagactactt taagcaagct tttccagaag gtttgtcctg
      361 ggaaagatct ttggaatttg aagatggtgg ttccgcttct gtttctgctc atatttcttt
      421 gagaggtaac accttctacc acaagtctaa gtttactggt gttaactttc cagccgatgg
      481 tccaattatg caaaatcaat ctgttgactg ggaaccatcc actgaaaaga ttactgcttc
      541 tgatggtgtt ttgaagggtg atgttactat gtacttgaag ttggaaggtg gtggtaacca
      601 taagtgtcaa ttcaagacta cttacaaggc cgccaaagaa attttggaaa tgccaggtga
      661 tcattacatc ggtcatagat tggttagaaa gaccgaaggt aacatcaccg aacaagttga
      721 agatgctgtt gctcattctt aaggcgcgcc acttctaaat aagcgaattt cttatgattt
      781 atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac
      841 tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg
      901 ctttctcagg tatagtatga ggtcgctctt attgaccaca cctctaccgg cagatccgct
      961 agggataaca gggtaatata gatctgttta gcttgcctcg tccccgccgg gtcacccggc
     1021 cagcgacatg gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc
     1081 atgatgtgac tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc
     1141 catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg
     1201 cgagcaggga aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta
     1261 gagaaatata aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa
     1321 aatcttgcta ggatacagtt ctcacatcac atccgaacat aaacaaccat gggtaaggaa
     1381 aagactcacg tttcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     1441 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag
     1501 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     1561 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     1621 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg caaaacagca
     1681 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     1741 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     1801 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     1861 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg
     1921 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     1981 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     2041 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     2101 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     2161 ctcgatgagt ttttctaatc agtactgaca ataaaaagat tcttgttttc aagaacttgt
     2221 catttgtata gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt
     2281 atattttttt tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa
     2341 tatcatgcgt caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac
     2401 gccgccatcc agtttaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct
     2461 atgcggccgc ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg
     2521 ttaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2581 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2641 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2701 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2761 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2821 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2881 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2941 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3001 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3061 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3121 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3181 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3241 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3301 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3361 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3421 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3481 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3541 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3601 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3661 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3721 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3781 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3841 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3901 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3961 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4021 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4081 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4141 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4201 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4261 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4321 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4381 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4441 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4501 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     4561 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     4621 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     4681 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     4741 agcagattgt actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt
     4801 aatacataac cttatgtatc atacacatac g
//
