LOCUS       sequence-401864-        7678 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrative vector for the expression of the yEmCherry-tagged 
            nuclear localizing 40nm nucGEMs under the INO4 promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7678)
  AUTHORS   Shu T et al.
  TITLE     nucGEMs probe the biophysical properties of the nucleoplasm
  JOURNAL   bioRxiv 2022
REFERENCE   2  (bases 1 to 7678)
  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: "bioRxiv 
            2022"
COMMENT     Sequence Label: pRS305-LEU2-pINO4-SV40NLS-PfV-GS-yEmCherry
FEATURES             Location/Qualifiers
     source          1..7678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..69)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     169..188
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(682..1776)
                     /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"
     promoter        complement(1777..2184)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(2485..2940)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2622..2643)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2834..2853
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3066..3088
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3080..3097
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3081..3097
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3104..3123
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3104..3122
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3313..3333
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5160..5347
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5215..5239)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        complement(5436..5454)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5475..5491)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5475..5491)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5488..5510)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5499..5515
                     /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(5523..5529,5530..5547,5548..5553))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5568..5589
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5706..5723)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5877..6465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5957..5976)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6636..7427,7428..7496))
                     /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     7259..7278
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7497..7601)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(7669..7678,1..9)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
       61 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
      121 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
      181 agattgtact gagagtgcac catatcgact acgtcgttaa ggccgtttct gacagagtaa
      241 aattcttgag ggaactttca ccattatggg aaatggttca agaaggtatt gacttaaact
      301 ccatcaaatg gtcaggtcat tgagtgtttt ttatttgttg tatttttttt tttttagaga
      361 aaatcctcca atatataaat taggaatcat agtttcatga ttttctgtta cacctaactt
      421 tttgtgtggt gccctcctcc ttgtcaatat taatgttaaa gtgcaattct ttttccttat
      481 cacgttgagc cattagtatc aatttgctta cctgtattcc tttacatcct cctttttctc
      541 cttcttgata aatgtatgta gattgcgtat atagtttcgt ctaccctatg aacatattcc
      601 attttgtaat ttcgtgtcgt ttctattatg aatttcattt ataaagttta tgtacaaata
      661 tcataaaaaa agagaatctt tttaagcaag gattttctta acttcttcgg cgacagcatc
      721 accgacttcg gtggtactgt tggaaccacc taaatcacca gttctgatac ctgcatccaa
      781 aaccttttta actgcatctt caatggcctt accttcttca ggcaagttca atgacaattt
      841 caacatcatt gcagcagaca agatagtggc gatagggttg accttattct ttggcaaatc
      901 tggagcagaa ccgtggcatg gttcgtacaa accaaatgcg gtgttcttgt ctggcaaaga
      961 ggccaaggac gcagatggca acaaacccaa ggaacctggg ataacggagg cttcatcgga
     1021 gatgatatca ccaaacatgt tgctggtgat tataatacca tttaggtggg ttgggttctt
     1081 aactaggatc atggcggcag aatcaatcaa ttgatgttga accttcaatg tagggaattc
     1141 gttcttgatg gtttcctcca cagtttttct ccataatctt gaagaggcca aaacattagc
     1201 tttatccaag gaccaaatag gcaatggtgg ctcatgttgt agggccatga aagcggccat
     1261 tcttgtgatt ctttgcactt ctggaacggt gtattgttca ctatcccaag cgacaccatc
     1321 accatcgtct tcctttctct taccaaagta aatacctccc actaattctc tgacaacaac
     1381 gaagtcagta cctttagcaa attgtggctt gattggagat aagtctaaaa gagagtcgga
     1441 tgcaaagtta catggtctta agttggcgta caattgaagt tctttacgga tttttagtaa
     1501 accttgttca ggtctaacac taccggtacc ccatttagga ccacccacag cacctaacaa
     1561 aacggcatca gccttcttgg aggcttccag cgcctcatct ggaagtggaa cacctgtagc
     1621 atcgatagca gcaccaccaa ttaaatgatt ttcgaaatcg aacttgacat tggaacgaac
     1681 atcagaaata gctttaagaa ccttaatggc ttcggctgtg atttcttgac caacgtggtc
     1741 acctggcaaa acgacgatct tcttaggggc agacattaga atggtatatc cttgaaatat
     1801 atatatatat attgctgaaa tgtaaaaggt aagaaaagtt agaaagtaag acgattgcta
     1861 accacctatt ggaaaaaaca ataggtcctt aaataatatt gtcaacttca agtattgtga
     1921 tgcaagcatt tagtcatgaa cgcttctcta ttctatatga aaagccggtt ccggcgctct
     1981 cacctttcct ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct
     2041 gaaattaaca aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg
     2101 ttctcgttat gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta
     2161 cgatacctga gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgcctt
     2221 agaccgctcg gccaaacaac caattacttg ttgagaaata gagtataatt atcctataaa
     2281 tataacgttt ttgaacacac atgaacaagg aagtacagga caattgattt tgaagagaat
     2341 gtggattttg atgtaattgt tgggattcca tttttaataa ggcaataata ttaggtatgt
     2401 agatatacta gaagttctcc tcgaccggtc gatatgcggt gtgaaatacc gcacagatgc
     2461 gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta aaattcgcgt
     2521 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     2581 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     2641 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     2701 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     2761 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     2821 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     2881 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     2941 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     3001 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     3061 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg actcactata
     3121 gggcgaattg gagctctctt ggcttggttt gtatgagctt gttacgtatc aaacaaaata
     3181 ttcacatgtt tttctcacct taaagacaga ggcgcgattg cgtttgcgaa attccaatgc
     3241 caacaagata agcttgacga aggagttaag agggcggctt gaactaaaaa gagaaaagca
     3301 atatctagaa tgccaaaaaa gaagagaaag gtcctctcaa taaatccaac ccttataaat
     3361 agggataagc catacaccaa agaagaatta atggaaattc tgagattagc aataatagct
     3421 gagcttgatg caataaatct atatgagcag atggcaagat attccgaaga tgaaaacgta
     3481 agaaagattc tccttgatgt tgcaagggag gagaaggctc acgttggaga attcatggca
     3541 ctccttctaa atttggatcc agaacaagta actgagctga agggtggatt tgaggaagtc
     3601 aaggagctaa ctggaattga ggctcacata aatgataaca agaaggagga atctaacgtg
     3661 gaatattttg agaagttgag aagtgctctt ttggatgggg taaataaggg aagatcttta
     3721 ctaaagcact tgccagtaac gagaatagaa ggacaatcat tcagggttga catcattaaa
     3781 tttgaagatg gtgttagggt agtaaagcaa gagtacaagc caattccatt attgaagaag
     3841 aaattctatg tgggaataag agaactcaac gatgggactt acgatgtatc tatagcaact
     3901 aaagctgggg aactcctagt taaagacgaa gagtcgctag taataagaga aattctatcc
     3961 acagagggaa ttaagaaaat gaaactttcc tcctgggaca atccagaaga agcacttaat
     4021 gatttaatga acgcactgca ggaagcctca aatgcgtcag ctggaccatt tgggctgata
     4081 attaacccaa aaagatatgc aaagttgcta aaaatttacg agaagagtgg aaagatgcta
     4141 gttgaagtgc taaaagagat attcagggga ggaatcattg ttactctaaa cattgatgag
     4201 aataaggtaa taatatttgc caatacacca gctgttcttg acgttgtagt tggccaagat
     4261 gtgactctac aagagctagg acctgaagga gacgacgttg cattcttggt tagtgaagct
     4321 ataggaataa gaataaagaa cccagaggca atagtagtct tagagtggtc tggcagcggt
     4381 agtgggtcgg gatcaggctc cggtcgacgg atccccgggt taattaagat ggtttcaaaa
     4441 ggtgaagaag ataatatggc tattattaaa gaatttatga gatttaaagt tcatatggaa
     4501 ggttcagtta atggtcatga atttgaaatt gaaggtgaag gtgaaggtag accatatgaa
     4561 ggtactcaaa ctgctaaatt gaaagttact aaaggtggtc cattaccatt tgcttgggat
     4621 attttgtcac cacaatttat gtatggttca aaagcttatg ttaaacatcc agctgatatt
     4681 ccagattatt taaaattgtc atttccagaa ggttttaaat gggaaagagt tatgaatttt
     4741 gaagatggtg gtgttgttac tgttactcaa gattcatcat tacaagatgg tgaatttatt
     4801 tataaagtta aattgagagg tactaatttt ccatcagatg gtccagttat gcaaaaaaaa
     4861 actatgggtt gggaagcttc atcagaaaga atgtatccag aagatggtgc tttaaaaggt
     4921 gaaattaaac aaagattgaa attaaaagat ggtggtcatt atgatgctga agttaaaact
     4981 acttataaag ctaaaaaacc agttcaatta ccaggtgctt ataatgttaa tattaaattg
     5041 gatattactt cacataatga agattatact attgttgaac aatatgaaag agctgaaggt
     5101 agacattcaa ctggtggtat ggatgaatta tataaataag gcgcgccact tctaaataag
     5161 cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat
     5221 acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt
     5281 tcctgtaggt caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct
     5341 ctaccggcag atccgctagg gataacaggg taatatagat ctgtttagct tgcctcgtcc
     5401 ccgctcgagg gggggcccgg taccagcttt tgttcccttt agtgagggtt aatttcgagc
     5461 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5521 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5581 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5641 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5701 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5761 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5821 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5881 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5941 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6001 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6061 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6121 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6181 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6241 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6301 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6361 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6421 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6481 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6541 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6601 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6661 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6721 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6781 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6841 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6901 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6961 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7021 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7081 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7141 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7201 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7261 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7321 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7381 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7441 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7501 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7561 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7621 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgta
//
