LOCUS       sequence-307770-        4900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast codon 
            optimized mTFP1 under KanMX selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4900)
  TITLE     Stelling Lab Plasmids
REFERENCE   2  (bases 1 to 4900)
  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: FRP265
FEATURES             Location/Qualifiers
     source          1..4900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      15..202
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(70..94)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            277..1633
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        277..620
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             621..1430
                     /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(688..707)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1436..1633
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1737..1756)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1738..1756)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1843..1860)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2014..2602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2094..2113)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2773..3564,3565..3633))
                     /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     3396..3415
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3634..3738)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3806..3824
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3862..3884)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3984..4003
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4084..4102
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4084..4101
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ccacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
       61 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat
      121 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagtat gaggtcgctc
      181 ttattgacca cacctctacc ggcagatccg ctagggataa cagggtaata tagatctgtt
      241 tagcttgcct cgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc
      301 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag
      361 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga
      421 agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac
      481 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca
      541 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc
      601 acatccgaac ataaacaacc atgggtaagg aaaagactca cgtttcgagg ccgcgattaa
      661 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
      721 caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac
      781 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga
      841 cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt
      901 tactcaccac tgcgatcccc ggcaaaacag cattccaggt attagaagaa tatcctgatt
      961 caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg
     1021 tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa
     1081 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
     1141 aacaagtctg gaaagaaatg cataagcttt tgccattctc accggattca gtcgtcactc
     1201 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
     1261 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
     1321 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
     1381 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa tcagtactga
     1441 caataaaaag attcttgttt tcaagaactt gtcatttgta tagttttttt atattgtagt
     1501 tgttctattt taatcaaatg ttagcgtgat ttatattttt tttcgcctcg acatcatctg
     1561 cccagatgcg aagttaagtg cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct
     1621 ggtcgctata ctgctgtcga ttcgatacta acgccgccat ccagtttaaa cgagctcgaa
     1681 ttcatcgatg atatcagatc cactagtggc ctatgcggcc gcggatctgc cggtctccct
     1741 atagtgagtc gtattaattt cgataagcca ggttaacctg cattaatgaa tcggccaacg
     1801 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     1861 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     1921 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     1981 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2041 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2101 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2161 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     2221 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2281 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2341 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2401 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     2461 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2521 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2581 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2641 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2701 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2761 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2821 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     2881 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     2941 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3001 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3061 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3121 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3181 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3241 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3301 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3361 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3421 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3481 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3541 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3601 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     3661 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3721 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     3781 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     3841 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     3901 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     3961 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     4021 ggacatattg tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat
     4081 acgatttagg tgacactata gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg
     4141 acggatcggt gacggtgctg gtttaattaa caccggtatg gtttctaaag gtgaagaaac
     4201 tactatgggt gttattaaac cagatatgaa gattaagttg aagatggaag gtaatgttaa
     4261 tggtcatgct tttgttattg aaggtgaagg tgaaggtaaa ccatatgatg gtacaaacac
     4321 tattaacttg gaagttaaag aaggtgctcc attgccattt tcttacgata ttttgactac
     4381 tgcttttgct tatggtaata gagcttttac taagtaccca gatgatattc caaattactt
     4441 caagcaatct tttccagaag gttattcttg ggaaagaact atgacttttg aagataaggg
     4501 tattgttaag gttaagtctg atatttctat ggaagaagat tctttcattt acgaaattca
     4561 tttgaagggt gaaaattttc caccaaatgg tccagttatg caaaaaaaga ctactggttg
     4621 ggatgcttct actgaaagga tgtacgtcag agatggtgtt ttgaaaggtg atgttaagca
     4681 taaattgttg ttggaaggtg gtggtcatca tagagttgat ttcaagacaa tttacagagc
     4741 taaaaaggct gttaaattgc cagattacca ttttgttgat catagaattg aaattttgaa
     4801 ccatgataag gattacaaca aggttactgt ttatgaatct gctgttgcta gaaattctac
     4861 tgatggtatg gatgaattgt ataaagccgg ctagggcgcg
//
