LOCUS       sequence-311210-        4888 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast codon 
            optimized mNeongreen under KanMX selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4888)
  TITLE     Stelling Lab Plasmids
REFERENCE   2  (bases 1 to 4888)
  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: FRP1975
FEATURES             Location/Qualifiers
     source          1..4888
                     /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      820..1007
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(875..899)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1082..2438
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1082..1425
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1426..2235
                     /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(1493..1512)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2241..2438
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2542..2561)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2543..2561)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2648..2665)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2819..3407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2899..2918)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3578..4369,4370..4438))
                     /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     4201..4220
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4439..4543)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4611..4629
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4667..4689)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4789..4808
                     /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 ggtgaagaag ataacatggc
      121 ttctttgcca gctactcatg aattgcatat tttcggttcc atcaacggtg ttgatttcga
      181 tatggttggt caaggtactg gtaatccaaa tgatggttac gaagaattga acttgaagtc
      241 cacaaaaggt gacttgcaat tctctccatg gattttggtt ccacatattg gttacggttt
      301 ccaccaatat ttgccatatc cagatggtat gtctccattt caagctgcta tggttgatgg
      361 ttctggttac caagttcata gaaccatgca atttgaagat ggtgcttctt tgaccgtcaa
      421 ctacagatat acttacgaag gttcccacat caaaggtgaa gctcaagtta agggtactgg
      481 ttttccagct gatggtccag ttatgactaa ttctttgact gctgctgatt ggtgcagatc
      541 caaaaaaact tacccaaacg ataagaccat catctctact ttcaagtggt cttacactac
      601 tggtaacggt aagagatata gatccactgc tagaactact tacactttcg ctaaacctat
      661 ggctgctaac tacttgaaga atcaacctat gtacgttttc agaaagaccg aattgaagca
      721 ctccaaaact gaattgaatt tcaaagaatg gcaaaaggcc ttcaccgatg ttatgggtat
      781 ggatgaatta tacaagtaag gcgcgccact tctaaataag cgaatttctt atgatttatg
      841 atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct
      901 taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt
      961 tctcaggtat agtatgaggt cgctcttatt gaccacacct ctaccggcag atccgctagg
     1021 gataacaggg taatatagat ctgtttagct tgcctcgtcc ccgccgggtc acccggccag
     1081 cgacatggag gcccagaata ccctccttga cagtcttgac gtgcgcagct caggggcatg
     1141 atgtgactgt cgcccgtaca tttagcccat acatccccat gtataatcat ttgcatccat
     1201 acattttgat ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc agacctgcga
     1261 gcagggaaac gctcccctca cagacgcgtt gaattgtccc cacgccgcgc ccctgtagag
     1321 aaatataaaa ggttaggatt tgccactgag gttcttcttt catatacttc cttttaaaat
     1381 cttgctagga tacagttctc acatcacatc cgaacataaa caaccatggg taaggaaaag
     1441 actcacgttt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     1501 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc
     1561 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     1621 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     1681 atccgtactc ctgatgatgc atggttactc accactgcga tccccggcaa aacagcattc
     1741 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     1801 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     1861 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     1921 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa gcttttgcca
     1981 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     2041 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     2101 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     2161 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     2221 gatgagtttt tctaatcagt actgacaata aaaagattct tgttttcaag aacttgtcat
     2281 ttgtatagtt tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata
     2341 ttttttttcg cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat
     2401 catgcgtcaa tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc
     2461 gccatccagt ttaaacgagc tcgaattcat cgatgatatc agatccacta gtggcctatg
     2521 cggccgcgga tctgccggtc tccctatagt gagtcgtatt aatttcgata agccaggtta
     2581 acctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2641 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2701 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2761 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2821 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2881 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2941 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3001 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3061 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3121 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3181 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3241 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3301 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3361 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3421 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3481 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3541 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3601 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3661 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3721 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3781 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3841 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3901 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3961 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4021 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4081 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4141 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4201 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4261 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4321 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4381 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4441 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4501 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4561 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     4621 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     4681 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     4741 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     4801 agattgtact gagagtgcac catatggaca tattgtcgtt agaacgcggc tacaattaat
     4861 acataacctt atgtatcata cacatacg
//
