LOCUS       sequence-311203-        4891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast codon 
            optimized mRuby2 under KanMX selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4891)
  TITLE     Stelling Lab Plasmids
REFERENCE   2  (bases 1 to 4891)
  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: FRP1931
FEATURES             Location/Qualifiers
     source          1..4891
                     /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             89..799
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     terminator      823..1010
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(878..902)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1085..2441
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1085..1428
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1429..2238
                     /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(1496..1515)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2244..2441
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2545..2564)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2546..2564)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2651..2668)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2822..3410)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2902..2921)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3581..4372,4373..4441))
                     /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     4204..4223
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4442..4546)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4614..4632
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4670..4692)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4792..4811
                     /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 aaacatgaga atgaaggtcg ttatggaagg ttctgttaac ggtcatcaat tcaagtgtac
      181 tggtgaaggt gaaggtaatc cttatatggg tactcaaacc atgagaatca aagtcattga
      241 aggtggtcca ttgccattcg cttttgatat tttggctacc tctttcatgt acggttccag
      301 aactttcatt aagtacccaa agggtatccc agacttcttc aaacaatctt ttccagaagg
      361 tttcacctgg gaaagagtta ctagatacga agatggtggt gttgttaccg ttatgcaaga
      421 tacttctttg gaagatggtt gcttggttta ccacgttcaa gttagaggtg ttaactttcc
      481 atctaacggt ccagtcatgc aaaaaaagac aaaaggttgg gaacctaaca ccgaaatgat
      541 gtatccagct gatggtggtt tgagaggtta tactcatatg gctttgaaag ttgacggtgg
      601 tggtcatttg tcttgttctt ttgttactac ctacagatcc aaaaagaccg tcggtaacat
      661 taagatgcca ggtattcatg ctgttgacca cagattggaa agattagaag aatccgacaa
      721 cgaaatgttc gtcgttcaaa gagaacacgc tgttgctaaa tttgctggtt tgggtggtgg
      781 tatggatgaa ttatacaagt gaggcgcgcc acttctaaat aagcgaattt cttatgattt
      841 atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac
      901 tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg
      961 ctttctcagg tatagtatga ggtcgctctt attgaccaca cctctaccgg cagatccgct
     1021 agggataaca gggtaatata gatctgttta gcttgcctcg tccccgccgg gtcacccggc
     1081 cagcgacatg gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc
     1141 atgatgtgac tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc
     1201 catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg
     1261 cgagcaggga aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta
     1321 gagaaatata aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa
     1381 aatcttgcta ggatacagtt ctcacatcac atccgaacat aaacaaccat gggtaaggaa
     1441 aagactcacg tttcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     1501 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag
     1561 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     1621 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     1681 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg caaaacagca
     1741 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     1801 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     1861 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     1921 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg
     1981 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     2041 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     2101 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     2161 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     2221 ctcgatgagt ttttctaatc agtactgaca ataaaaagat tcttgttttc aagaacttgt
     2281 catttgtata gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt
     2341 atattttttt tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa
     2401 tatcatgcgt caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac
     2461 gccgccatcc agtttaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct
     2521 atgcggccgc ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg
     2581 ttaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2641 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2701 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2761 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2821 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2881 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2941 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3001 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3061 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3121 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3181 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3241 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3301 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3361 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3421 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3481 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3541 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3601 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3661 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3721 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3781 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3841 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3901 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3961 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4021 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4081 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4141 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4201 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4261 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4321 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4381 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4441 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4501 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4561 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     4621 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     4681 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     4741 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     4801 agcagattgt actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt
     4861 aatacataac cttatgtatc atacacatac g
//
