LOCUS       sequence-106469-        3724 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector to create a cassette for homologous recombination in S. 
            cerevisiae; adds a C-terminal Strep tag II followed by TRP1 
            selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3724)
  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 3724)
  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: pGS1011
FEATURES             Location/Qualifiers
     source          1..3724
                     /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             423..446
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(481..497)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(481..497)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1503..1519)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1503..1519)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1516..1538)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1527..1543
                     /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(1551..1557,1558..1575,1576..1581))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1596..1617
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1734..1751)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1905..2493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1985..2004)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2664..3455,3456..3524))
                     /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     3287..3306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3525..3629)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3697..3715
                     /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 accggtagtg
      421 cttggagtca tccacaattc gaaaaataaa agcttgatat cgaattcctg cagcccgggg
      481 gatccactag ttctagagcg gccgcggaac gatcattcac tatatatata tcaatttata
      541 tatacgtatg tgtaattgaa gaaagatacg ttttttcctc tattgagagg cctgctggat
      601 gaatagcttt accttttcta aatccttgat accatcagtc tctactcctc cacttacatc
      661 gacaccgatc gcatttggta acatattaat ggcaacagaa acgttatcag gattcaatcc
      721 accagcgata atgaatttta tctcgggatg acttgcagac caactggaaa ttgcactcca
      781 attcaatttc tcaccagtgc caccttcacc agaatcgaac aacgtcagca cattgtctac
      841 gtgttcatac aggtccagta gtaattcaca atcctgtggg aactggaacc tcttaatgat
      901 tggaattgaa gatgggatca aagatctgta ttctttaata tcttcatctc catgtaattg
      961 tatcacatct agattatatt cgtggtacag ttgaaggaca tcatcaacgg actgatttct
     1021 aaacaccccg actagtttag tacctttcac gttctcttgt tggtgaactg cagttgaaat
     1081 acctttcgca acagatgagt caatggttct tttcctaccg ggaacacaaa tgatacctaa
     1141 gtaatcagca ccatcatcca cagcagtctt tgcagcttca acggtttgca aaccacacac
     1201 tttaacgagc atcacagtag atttcgcaag agggtagctc gctcagagta cccaagtaaa
     1261 tgattagtaa actgatgttt gatagttcaa tttttcaatg aaataacctt atattaaaat
     1321 tgatattact attatacaaa aataaagaat aaaggatttg agtttataca taaaatacca
     1381 ttattatttg ttcagtgaga gataccgggg tatatgggat gtgtgtagtg ataccatgca
     1441 atcatgtatc aaacatgggc cctcgctccg accgtgcggc atgcaagctt ggcgtaatca
     1501 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1561 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1621 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     1681 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     1741 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     1801 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1861 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1921 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1981 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2041 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2101 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2161 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2221 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2281 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2341 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2401 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2461 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2521 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2581 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2641 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2701 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2761 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2821 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2881 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     2941 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3001 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3061 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3121 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3181 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3241 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3301 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3361 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3421 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3481 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3541 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3601 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     3661 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     3721 cgtc
//
