LOCUS       sequence-92983-e        4287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector to create a cassette for homologous recombination in S. 
            cerevisiae; adds a C-terminal Twin-Strep tag and a URA3 selection 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4287)
  AUTHORS   Rai J, Pemmasani JK, Voronovsky A, Jensen IS, Manavalan A, Nyengaard
            JR, Golas MM, Sander B
  TITLE     Strep-tag II and Twin-Strep Based Cassettes for Protein Tagging by 
            Homologous Recombination and Characterization of Endogenous 
            Macromolecular Assemblies in Saccharomyces cerevisiae.
  JOURNAL   Mol Biotechnol. 2014 Jun 27.
  PUBMED    24969434
REFERENCE   2  (bases 1 to 4287)
  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"; journalName: "Mol 
            Biotechnol. 2014 Jun 27."
COMMENT     Sequence Label: pGS1022
FEATURES             Location/Qualifiers
     source          1..4287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             462..485
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             522..545
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(581..597)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(581..597)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      complement(636..833)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(1652..1995)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     complement(2066..2082)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2066..2082)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2079..2101)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2090..2106
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2114..2120,2121..2138,2139..2144))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2159..2180
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2297..2314)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2468..3056)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2548..2567)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3227..4018,4019..4087))
                     /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     3850..3869
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4088..4192)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4260..4278
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccggggat
      421 ccctcgaggt cgacctgcag ggggaccatg gtctcagcgc ttggtcgcat ccacaatttg
      481 agaaaggtgg aggttccgga ggtggatcgg gaggtggatc gtggagccac ccgcagttcg
      541 aaaaataata agcttgatat cgaattcctg cagcccgggg gatccactag ttctagagcg
      601 gccgctggat ggcggcgtta gtatcgaatc gacagcagta tagcgaccag cattcacata
      661 cgattgacgc atgatattac tttctgcgca cttaacttcg catctgggca gatgatgtcg
      721 aggcgaaaaa aaatataaat cacgctaaca tttgattaaa atagaacaac tacaatataa
      781 aaaaactata caaatgacaa gttcttgaaa acaagaatct ttttattgtc agtactgatt
      841 ataattggcc agtctttttc aaataagcat tccaaccagc atctctatac cttttacctt
      901 caatatctgg atctcttcct ttaccaaaca atcctctacc aacaatgata atatcagttc
      961 cagtgctaac aacttcatca acagttctat attgttgtcc taatccatca cctttatcat
     1021 ctaatccaac tccaggtgtc ataataagcc aatcaaatcc ttcttcttgt ccacccatat
     1081 cacgttgggc aataaatcca ataacaaatt ccttatcgga tttagcaatt tcaacagttt
     1141 tttgagaata ttctccatat gctaatgatc ccactgatga taattcagct aacatcaata
     1201 accctcttgg ctcttggttg gtggtggttt ctttagctcc ctgttttaat ccttcaacta
     1261 ctccattccc agtgacacca tgagcattag taatatctgc ccaactacta attttataaa
     1321 ctccaccaat atattgtttc ttcacggtat taccaatatc agcaaatttt ctatcttcaa
     1381 aaatcataaa ttgatgttta cgtgaaagtt ctaataatgg ttcaatagtg gattcatagg
     1441 aaaaatcatt gattatatca atatgagtct tgattaagca tacataagga cccaatttat
     1501 caattaattc aaggaattcc ttagtggtat caacatcaat tgatgcacat aaattggttt
     1561 tcttcagttc cattaatcga aataatcgtt gtgctactgg tgaggcatga gtttctgctc
     1621 tctcactata ggtcttagtg ttgactgtca tggttgttta tgttcggatg tgatgtgaga
     1681 actgtatcct agcaagattt taaaaggaag tatatgaaag aagaacctca gtggcaaatc
     1741 ctaacctttt atatttctct acaggggcgc ggcgtgggga caattcaacg cgtctgtgag
     1801 gggagcgttt ccctgctcgc aggtccgcag cgaggagccg taatttttgc ttcgcgccgt
     1861 gcggccatca aaatgtatgg atgcaaatga ttatacatgg ggatgtatgg gctaaatgta
     1921 cgggcgacag tcacatcatg cccctgagct gcgcacgtca agactgtcaa ggagggtatt
     1981 ctgggcctcc atgtcgctgg ccgggtgacc cggcggggac gaggcaagct gtcgatcagg
     2041 cgcatgcaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     2101 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     2161 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     2221 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     2281 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2341 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2401 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2461 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2521 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2581 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2641 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2701 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2761 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2821 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2881 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     2941 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3001 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3061 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3121 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3181 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3241 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3301 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3361 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3421 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3481 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3541 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3601 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     3661 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     3721 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     3781 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     3841 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     3901 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     3961 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4021 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4081 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4141 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4201 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     4261 ataggcgtat cacgaggccc tttcgtc
//
