LOCUS       sequence-311209-        4912 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast codon 
            optimized mNeptune2 under KanMX selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4912)
  TITLE     Stelling Lab Plasmids
REFERENCE   2  (bases 1 to 4912)
  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: FRP2000
FEATURES             Location/Qualifiers
     source          1..4912
                     /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      844..1031
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(899..923)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1106..2462
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1106..1449
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1450..2259
                     /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(1517..1536)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2265..2462
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2566..2585)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2567..2585)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2672..2689)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2843..3431)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2923..2942)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3602..4393,4394..4462))
                     /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     4225..4244
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4463..4567)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4635..4653
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4691..4713)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4813..4832
                     /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 tgatcaaaga
      121 aaacatgcac atgaagttgt acatggaagg tactgttaac aaccatcatt tcaagtgtac
      181 ctctgaaggt gaaggtaaac catatgaagg tactcaaact ggtagaatca aggttgttga
      241 aggtggtcca ttgccatttg cttttgatat tttggctacc tgcttcatgt acggttctaa
      301 gactttcatt aaccacaccc aaggtatccc agatttcttc aaacaatctt tcccagaagg
      361 tttcacttgg gaaagagtta ctacttacga agatggtggt gttttgactg ttactcaaga
      421 tacctcttta caagacggtt gcttgatcta caacgttaag ttgagaggtg ttaacttccc
      481 atctaatggt ccagtcatgc aaaaaaagac tttgggttgg gaagctagta ctgaaacatt
      541 atatccagct gatggtggtt tggaaggtag atgtgatatg gctttgaaat tggttggtgg
      601 tggtcacttg cattgcaact tgaaaactac ttacagatcc aagaagccag ccaagaattt
      661 gaaaatgcca ggtgtttact tcgtcgatag aagattggaa agaatcaaag aagccgacaa
      721 cgaaacctac gttgaacaac acgaagttgc tgttgctaga tattgtgatt tgccatctaa
      781 gttgggtcac aagttgaatg gtatggacga attatacaag taaggcgcgc cacttctaaa
      841 taagcgaatt tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt
      901 gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac
      961 tctttcctgt aggtcaggtt gctttctcag gtatagtatg aggtcgctct tattgaccac
     1021 acctctaccg gcagatccgc tagggataac agggtaatat agatctgttt agcttgcctc
     1081 gtccccgccg ggtcacccgg ccagcgacat ggaggcccag aataccctcc ttgacagtct
     1141 tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg tacatttagc ccatacatcc
     1201 ccatgtataa tcatttgcat ccatacattt tgatggccgc acggcgcgaa gcaaaaatta
     1261 cggctcctcg ctgcagacct gcgagcaggg aaacgctccc ctcacagacg cgttgaattg
     1321 tccccacgcc gcgcccctgt agagaaatat aaaaggttag gatttgccac tgaggttctt
     1381 ctttcatata cttcctttta aaatcttgct aggatacagt tctcacatca catccgaaca
     1441 taaacaacca tgggtaagga aaagactcac gtttcgaggc cgcgattaaa ttccaacatg
     1501 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     1561 atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     1621 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     1681 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     1741 gcgatccccg gcaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     1801 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     1861 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     1921 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     1981 aaagaaatgc ataagctttt gccattctca ccggattcag tcgtcactca tggtgatttc
     2041 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     2101 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     2161 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     2221 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagtactgac aataaaaaga
     2281 ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt
     2341 aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga
     2401 agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac
     2461 tgctgtcgat tcgatactaa cgccgccatc cagtttaaac gagctcgaat tcatcgatga
     2521 tatcagatcc actagtggcc tatgcggccg cggatctgcc ggtctcccta tagtgagtcg
     2581 tattaatttc gataagccag gttaacctgc attaatgaat cggccaacgc gcggggagag
     2641 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2701 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2761 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2821 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2881 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2941 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3001 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3061 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3121 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3181 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3241 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3301 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3361 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3421 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3481 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3541 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3601 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3661 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3721 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3781 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3841 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3901 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3961 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4021 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4081 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4141 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4201 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4261 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4321 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4381 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4441 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4501 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4561 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     4621 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     4681 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     4741 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     4801 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg gacatattgt
     4861 cgttagaacg cggctacaat taatacataa ccttatgtat catacacata cg
//
