LOCUS       sequence-451056-        7263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gal1p dependent budding yeast expression of tandem-affinity tagged 
            spt6p tSH2 domain (1123-1451) localized to nucleus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7263)
  TITLE     Hill lab spt6-tsh2 plasmids
REFERENCE   2  (bases 1 to 7263)
  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: CPH3375_SF_Spt6p-tSH2-1223-1451_NLS
FEATURES             Location/Qualifiers
     source          1..7263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        14..678
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    230..347
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     642..665
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             694..717
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             733..756
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             793..816
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1516..1536
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1594..1610)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1594..1610)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1607..1629)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1618..1634
                     /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(1642..1648,1649..1666,1667..1672))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1687..1708
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1825..1842)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1996..2584)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2076..2095)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2755..3546,3547..3615))
                     /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     3378..3397
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3616..3720)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3788..3806
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3826..4231
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             4232..5326
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     rep_origin      5464..6810
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     7232..7254
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7246..7263
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7247..7263
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gaattccttg aattttcaaa aattcttact ttttttttgg atggacgcaa agaagtttaa
       61 taatcatatt acatggcatt accaccatat acatatccat atacatatcc atatctaatc
      121 ttacttatat gttgtggaaa tgtaaagagc cccattatct tagcctaaaa aaaccttctc
      181 tttggaactt tcagtaatac gcttaactgc tcattgctat attgaagtac ggattagaag
      241 ccgccgagcg ggtgacagcc ctccgaagga agactctcct ccgtgcgtcc tcgtcttcac
      301 cggtcgcgtt cctgaaacgc agatgtgcct cgcgccgcac tgctccgaac aataaagatt
      361 ctacaatact agcttttatg gttatgaaga ggaaaaattg gcagtaacct ggccccacaa
      421 accttcaaat gaacgaatca aattaacaac cataggatga taatgcgatt agttttttag
      481 ccttatttct ggggtaatta atcagcgaag cgatgatttt tgatctatta acagatatat
      541 aaatgcaaaa actgcataac cactttaact aatactttca acattttcgg tttgtattac
      601 ttcttattca aatgtaataa aagtatcaac aaaaaattgt taatatacct ctatacttta
      661 acgtcaagga gaaaaaaccc cggatctcat atggactaca aggatgacga cgacaagggt
      721 accgccgcca gctggagcca cccgcagttc gaaaaaggcg gcggcagcgg cggcggcagc
      781 ggcggcggca gctggagcca cccgcagttc gaaaaaggct cgagcatttg ggacttgaaa
      841 caagaactgg aagatgccga agaggagagg aaattgatga tggcagaagc ccgtgcaaag
      901 agaacacatc gtgttatcaa tcatccttac tatttccctt tcaacggcag acaggctgag
      961 gattacttaa ggagtaaaga acgtggtgaa ttcgtgatca gacagtctag ccgaggtgat
     1021 gaccacttgg ttatcacctg gaaattggat aaggatttgt ttcaacatat tgatatccaa
     1081 gaattagaaa aagaaaatcc tttggcttta ggtaaagtct tgattgtcga caatcagaaa
     1141 tacaatgatt tagaccagat cattgtagaa tatcttcaaa acaaggtaag gctcttgaat
     1201 gaaatgacat ctagtgaaaa attcaaaagc ggtactaaga aagatgtggt caagtttatt
     1261 gaagactact ctagagtgaa tccaaataag tctgtgtact atttcagttt gaaccacgat
     1321 aaccctggtt ggttttactt gatgttcaag attaacgcaa atagcaaatt atacacatgg
     1381 aatgtgaaat taacgaacac tggttatttc ctggtaaact acaattatcc aagtgttatc
     1441 cagctttgta atggttttaa gacgcttcta aaatctaaca gtagtaagaa tagaatgaac
     1501 aactaccgtg gagggccaaa aaagaagaga aaggtcggag ggtaaggatc ctctagagtc
     1561 gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     1621 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     1681 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     1741 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1801 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1861 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1921 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1981 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2041 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2101 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2161 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2221 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2281 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2341 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2401 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2461 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2521 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2581 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2641 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2701 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2761 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2821 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2881 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     2941 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3001 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3061 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3121 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3181 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3241 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3301 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3361 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3421 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3481 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3541 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3601 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3661 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3721 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     3781 ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaattaactg tgggaatact
     3841 caggtatcgt aagatgcaag agttcgaatc tcttagcaac cattattttt ttcctcaaca
     3901 taacgagaac acacaggggc gctatcgcac agaatcaaat tcgatgactg gaaatttttt
     3961 gttaatttca gaggtcgcct gacgcatata cctttttcaa ctgaaaaatt gggagaaaaa
     4021 ggaaaggtga gagcgccgga accggctttt catatagaat agagaagcgt tcatgactaa
     4081 atgcttgcat cacaatactt gaagttgaca atattattta aggacctatt gttttttcca
     4141 ataggtggtt agcaatcgtc ttactttcta acttttctta ccttttacat ttcagcaata
     4201 tatatatata tttcaaggat ataccattct aatgtctgcc cctaagaaga tcgtcgtttt
     4261 gccaggtgac cacgttggtc aagaaatcac agccgaagcc attaaggttc ttaaagctat
     4321 ttctgatgtt cgttccaatg tcaagttcga tttcgaaaat catttaattg gtggtgctgc
     4381 tatcgatgct acaggtgttc cacttccaga tgaggcgctg gaagcctcca agaaggctga
     4441 tgccgttttg ttaggtgctg tgggtggtcc taaatggggt acaggtagtg ttagacctga
     4501 acaaggttta ctaaaaatcc gtaaagaact tcaattgtac gccaacttaa gaccatgtaa
     4561 ctttgcatcc gactctcttt tagacttatc tccaatcaag ccacaatttg ctaaaggtac
     4621 tgacttcgtt gttgtcagag aattagtggg aggtatttac tttggtaaga gaaaggaaga
     4681 cgatggtgat ggtgtcgctt gggatagtga acaatacacc gttccagaag tgcaaagaat
     4741 cacaagaatg gccgctttca tggccctaca acatgagcca ccattgccta tttggtcctt
     4801 ggataaagct aatgttttgg cctcttcaag attatggaga aaaactgtgg aggaaaccat
     4861 caagaacgaa tttcctacat tgaaggttca acatcaattg attgattctg ccgccatgat
     4921 cctagttaag aacccaaccc acctaaatgg tattataatc accagcaaca tgtttggtga
     4981 tatcatctcc gatgaagcct ccgttatccc aggttccttg ggtttgttgc catctgcgtc
     5041 cttggcctct ttgccagaca agaacaccgc atttggtttg tacgaaccat gccacggttc
     5101 tgctccagat ttgccaaaga ataaggtcaa ccctatcgcc actatcttgt ctgctgcaat
     5161 gatgttgaaa ttgtcattga acttgcctga agaaggtaag gccattgaag atgcagttaa
     5221 aaaggttttg gatgcaggta tcagaactgg tgatttaggt ggttccaaca gtaccaccga
     5281 agtcggtgat gctgtcgccg aagaagttaa gaaaatcctt gcttaaaaag attctctttt
     5341 tttatgatat ttgtacataa actttataaa tgaaattcat aatagaaacg acacgaaatt
     5401 acaaaatgga atatgttcat agggtagaat taattctcat gtttgacagc ttatcatcgg
     5461 atcgatccaa tatcaaagga aatgatagca ttgaaggatg agactaatcc aattgaggag
     5521 tggcagcata tagaacagct aaagggtagt gctgaaggaa gcatacgata ccccgcatgg
     5581 aatgggataa tatcacagga ggtactagac tacctttcat cctacataaa tagacgcata
     5641 taagtacgca tttaagcata aacacgcact atgccgttct tctcatgtat atatatatac
     5701 aggcaacacg cagatatagg tgcgacgtga acagtgagct gtatgtgcgc agctcgcgtt
     5761 gcattttcgg aagcgctcgt tttcggaaac gctttgaagt tcctattccg aagttcctat
     5821 tctctagcta gaaagtatag gaacttcaga gcgcttttga aaaccaaaag cgctctgaag
     5881 acgcactttc aaaaaaccaa aaacgcaccg gactgtaacg agctactaaa atattgcgaa
     5941 taccgcttcc acaaacattg ctcaaaagta tctctttgct atatatctct gtgctatatc
     6001 cctatataac ctacccatcc acctttcgct ccttgaactt gcatctaaac tcgacctcta
     6061 cattttttat gtttatctct agtattactc tttagacaaa aaaattgtag taagaactat
     6121 tcatagagtg aatcgaaaac aatacgaaaa tgtaaacatt tcctatacgt agtatataga
     6181 gacaaaatag aagaaaccgt tcataatttt ctgaccaatg aagaatcatc aacgctatca
     6241 ctttctgttc acaaagtatg cgcaatccac atcggtatag aatataatcg gggatgcctt
     6301 tatcttgaaa aaatgcaccc gcagcttcgc tagtaatcag taaacgcggg aagtggagtc
     6361 aggctttttt tatggaagag aaaatagaca ccaaagtagc cttcttctaa ccttaacgga
     6421 cctacagtgc aaaaagttat caagagactg cattatagag cgcacaaagg agaaaaaaag
     6481 taatctaaga tgctttgtta gaaaaatagc gctctcggga tgcatttttg tagaacaaaa
     6541 aagaagtata gattctttgt tggtaaaata gcgctctcgc gttgcatttc tgttctgtaa
     6601 aaatgcagct cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat ttttgtttta
     6661 caaaaatgaa gcacagattc ttcgttggta aaatagcgct ttcgcgttgc atttctgttc
     6721 tgtaaaaatg cagctcagat tctttgtttg aaaaattagc gctctcgcgt tgcatttttg
     6781 ttctacaaaa tgaagcacag atgcttcgtt aacaaagata tgctattgaa gtgcaagatg
     6841 gaaacgcaga aaatgaaccg gggatgcgac gtgcaagatt acctatgcaa tagatgcaat
     6901 agtttctcca ggaaccgaaa tacatacatt gtcttccgta aagcgctaga ctatatatta
     6961 ttatacaggt tcaaatatac tatctgtttc agggaaaact cccaggttcg gatgttcaaa
     7021 attcaatgat gggtaacaag tacgatccga taaatgcggt gtgaaatacc gcacagatgc
     7081 gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa
     7141 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     7201 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     7261 agt
//
