LOCUS       sequence-311204-        5554 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast codon 
            optimized tdKOkappa under KanMX selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5554)
  TITLE     Stelling Lab Plasmids
REFERENCE   2  (bases 1 to 5554)
  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: FRP1864
FEATURES             Location/Qualifiers
     source          1..5554
                     /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      1486..1673
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1541..1565)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1748..3104
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1748..2091
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2092..2901
                     /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(2159..2178)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2907..3104
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3208..3227)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3209..3227)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3314..3331)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3485..4073)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3565..3584)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4244..5035,5036..5104))
                     /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     4867..4886
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5105..5209)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5277..5295
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5333..5355)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5455..5474
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atttaggtga cactatagaa cgcggccgcc agctgaagct tcgtacgctg caggtcgacg
       61 gatcggtgac ggtgctggtt taattaacat ggtcagtgtc atcaagccag aaatgaagat
      121 gagatattac atggacggtt ctgttaacgg tcatgaattc actattgaag gtgaaggtac
      181 tggtagacca tatgaaggtc atcaagaaat gactttgaga gttactatgg ctgaaggtgg
      241 tccaatgcca tttgcttttg atttggtttc tcacgttttc tgctacggtc atagagtttt
      301 tactaagtac ccagaagaaa tcccagacta ctttaagcaa gcttttccag aaggtttgtc
      361 ctgggaaaga tcattggaat ttgaagatgg tggttccgct tctgtttctg ctcatatttc
      421 tttgagaggt aacaccttct accacaagtc taagtttact ggtgttaact ttccagccga
      481 tggtccaatt atgcaaaatc aatctgttga ctgggaacca tccactgaaa agattactgc
      541 ttctgatggt gttttgaagg gtgatgttac tatgtacttg aagttggaag gtggtggtaa
      601 ccataagtgt caattcaaga ctacttacaa ggccgccgaa aagattttgg aaatgccagg
      661 tgatcattac atcggtcaca gattggttag aaagaccgaa ggtaacatta ccgaacaagt
      721 tgaagatgct gttgctcatt ctcatggtac tggttctaca ggttctggtt cttctggtac
      781 agcttcttct gaagataaca acatggctat ggtttccgtt attaagcctg aaatgaagat
      841 gagatattac atggatggta gtgtcaatgg tcacgaattc acaatagaag gtgaaggtac
      901 aggtagacct tacgaaggtc accaagaaat gacattaaga gtaacaatgg ccgaaggtgg
      961 tcctatgcct ttcgcattcg acttagtttc acatgtcttt tgctatggtc acagagtatt
     1021 caccaaatat cctgaagaaa ttcctgatta cttcaaacaa gccttccctg aaggtttaag
     1081 ttgggaaaga agtttagaat tcgaagatgg tggtagtgct tcagttagtg cacatattag
     1141 tttaagaggt aatacttttt atcataagtc caagttcacc ggtgtcaatt tccctgctga
     1201 tggtcctata atgcaaaatc aaagtgtaga ttgggaacct tctacagaaa aaatcacagc
     1261 atcagacggt gtattgaaag gtgacgtaac catgtatttg aaattagaag gtggtggtaa
     1321 tcacaaatgc caattcaaaa caacctacaa agctgctgaa aaaatcttag aaatgcctgg
     1381 tgaccactac attggtcata gattagtcag aaaaactgaa ggtaatatca ctgaacaagt
     1441 cgaagatgca gtagcccatt cataaggcgc gccacttcta aataagcgaa tttcttatga
     1501 tttatgattt ttattattaa ataagttata aaaaaaataa gtgtatacaa attttaaagt
     1561 gactcttagg ttttaaaacg aaaattctta ttcttgagta actctttcct gtaggtcagg
     1621 ttgctttctc aggtatagta tgaggtcgct cttattgacc acacctctac cggcagatcc
     1681 gctagggata acagggtaat atagatctgt ttagcttgcc tcgtccccgc cgggtcaccc
     1741 ggccagcgac atggaggccc agaataccct ccttgacagt cttgacgtgc gcagctcagg
     1801 ggcatgatgt gactgtcgcc cgtacattta gcccatacat ccccatgtat aatcatttgc
     1861 atccatacat tttgatggcc gcacggcgcg aagcaaaaat tacggctcct cgctgcagac
     1921 ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct
     1981 gtagagaaat ataaaaggtt aggatttgcc actgaggttc ttctttcata tacttccttt
     2041 taaaatcttg ctaggataca gttctcacat cacatccgaa cataaacaac catgggtaag
     2101 gaaaagactc acgtttcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     2161 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg
     2221 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     2281 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     2341 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggcaaaaca
     2401 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     2461 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     2521 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     2581 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataagctt
     2641 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     2701 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     2761 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     2821 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     2881 atgctcgatg agtttttcta atcagtactg acaataaaaa gattcttgtt ttcaagaact
     2941 tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat gttagcgtga
     3001 tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt gcgcagaaag
     3061 taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg attcgatact
     3121 aacgccgcca tccagtttaa acgagctcga attcatcgat gatatcagat ccactagtgg
     3181 cctatgcggc cgcggatctg ccggtctccc tatagtgagt cgtattaatt tcgataagcc
     3241 aggttaacct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3301 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3361 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3421 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3481 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3541 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3601 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3661 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3721 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3781 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3841 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3901 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3961 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4021 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4081 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4141 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4201 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4261 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4321 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4381 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4441 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4501 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4561 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4621 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4681 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4741 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4801 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4861 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4921 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4981 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5041 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5101 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5161 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5221 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     5281 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     5341 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     5401 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     5461 cagagcagat tgtactgaga gtgcaccata tggacatatt gtcgttagaa cgcggctaca
     5521 attaatacat aaccttatgt atcatacaca tacg
//
