LOCUS       sequence-367566-        8984 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NUP1-HA CEN TRP1 AmpR. Yeast shuttle plasmid for expression of 
            hemagluttinin tagged NUP1 nucleoporin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8984)
  AUTHORS   Belanger KD, Kenna MA, Wei S, Davis LI
  TITLE     Genetic and physical interactions between Srp1p and nuclear pore 
            complex proteins Nup1p and Nup2p.
  JOURNAL   J Cell Biol. 1994 Aug;126(3):619-30. doi: 10.1083/jcb.126.3.619.
  PUBMED    8045927
REFERENCE   2  (bases 1 to 8984)
  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"; doi: "10.1083/jcb.126.3";
            journalName: "J Cell Biol"; date: "1994-08"; volume: "126"; issue: 
            "3"; pages: "619-30"
COMMENT     Sequence Label: LDB107
FEATURES             Location/Qualifiers
     source          1..8984
                     /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"
     promoter        191..471
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             472..1146
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(1249..1704)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1386..1407)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1598..1617
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1830..1852
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1844..1861
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1845..1861
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1868..1887
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1868..1886
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1919..1935
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1919..1935
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             complement(4192..4218)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(6166..6182)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6167..6183)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(6211..6229)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6250..6266)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6250..6266)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6263..6285)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6274..6290
                     /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(6298..6304,6305..6322,6323..6328))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6343..6364
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6481..6498)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6652..7240)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6732..6751)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7411..8202,8203..8271))
                     /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     8034..8053
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8272..8376)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    8413..8916
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     8957..8975
                     /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 accatagatc acgacattac tatatatata atataggaag catttaatag aacagcatcg
      241 taatatatgt gtactttgca gttatgacgc cagatggcag tagtggaaga tattctttat
      301 tgaaaaatag cttgtcacct tacgtacaat cttgatccgg agcttttctt tttttgccga
      361 ttaagaatta attcggtcga aaaaagaaaa ggagagggcc aagagggagg gcattggtga
      421 ctattgagca cgtgagtata cgtgattaag cacacaaagg cagcttggag tatgtctgtt
      481 attaatttca caggtagttc tggtccattg gtgaaagttt gcggcttgca gagcacagag
      541 gccgcagaat gtgctctaga ttccgatgct gacttgctgg gtattatatg tgtgcccaat
      601 agaaagagaa caattgaccc ggttattgca aggaaaattt caagtcttgt aaaagcatat
      661 aaaaatagtt caggcactcc gaaatacttg gttggcgtgt ttcgtaatca acctaaggag
      721 gatgttttgg ctctggtcaa tgattacggc attgatatcg tccaactgca tggagatgag
      781 tcgtggcaag aataccaaga gttcctcggt ttgccagtta ttaaaagact cgtatttcca
      841 aaagactgca acatactact cagtgcagct tcacagaaac ctcattcgtt tattcccttg
      901 tttgattcag aagcaggtgg gacaggtgaa cttttggatt ggaactcgat ttctgactgg
      961 gttggaaggc aagagagccc cgaaagctta cattttatgt tagctggtgg actgacgcca
     1021 gaaaatgttg gtgatgcgct tagattaaat ggcgttattg gtgttgatgt aagcggaggt
     1081 gtggagacaa atggtgtaaa agactctaac aaaatagcaa atttcgtcaa aaatgctaag
     1141 aaataggtta ttactgagta gtatttattt aagtattgtt tgtgcacttg ccgatctatg
     1201 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg
     1261 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     1321 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     1381 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     1441 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     1501 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     1561 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     1621 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     1681 ttaatgcgcc gctacagggc gcgtccattc gccattcagg ctgcgcaact gttgggaagg
     1741 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     1801 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     1861 tgaattgtaa tacgactcac tatagggcga attggagctc caccgcggtg gcggccgctc
     1921 tagaactagt ggatcccccg ggctgcagga attcctgcag cccctgatcg aagtaaacag
     1981 aaaagaacac taattgtaga tcaggctgtg tactagacct tattttactg tattttttcg
     2041 gaaagaaaaa aggagcgctt tgcagatcga aagtttcgct cgtaaattat ttgtaagatg
     2101 ctattcataa tatgttaact gagagaaacc aggtcaaaac aaaacaattt tgggctcttg
     2161 cctccaaatt tgcctaccct agaacaggta tccattatct cgcctgtacc cgattaaaaa
     2221 aaagaccaat tatttaaaac ttctcaagaa gtttcatatg cagtgtataa gttgaaggaa
     2281 tataggaata tatatccttc agaaaagcaa cacaatacct aattacataa ccgatattta
     2341 ccttttagag tgcctcattc ttgcaatctt tctgttcgcc ataacaccac cgcccatgct
     2401 catgccatta tttgttccca tccccatctg attaggggct gactgcggct gcccaaaaga
     2461 agttgtcggc acaccacctg cccccccaaa aatggatgat ggatttgtta cgtttgaatt
     2521 ggaaccagag gcagcattcg caccaaatat atcacttggc cttaatgcat tggtcgcggt
     2581 attagtaatt ccgccattca atccgctaaa attaatatta ggaactgttg atggcgtgaa
     2641 tgagctgttt gtattgaaag atggggtttg cgattgatgt ggttggttat tagagccggc
     2701 aaacaccgta ttagcattag tgttgccgtt catattaaat acagagccgc caccaggcgt
     2761 tgaattattt cccgtaaaat taaaagcaga gggaacattg acattttgtg cattcgtgga
     2821 aggaggttta ttgaataaac cagcattagc attagttgac gaagttgctg atgtaaaagg
     2881 attgagaccc cctgcaccat tgtttccaaa attaaatgaa gattgattag aagctgcacc
     2941 agttgctgct gttcctgagc tcgaaaagcc aaatgccgag cccgctccat tcgtattgcc
     3001 actagcgata ctttgatccg gttttcctac gttaaatgta cccgctatat tggttcctga
     3061 ggtattggaa gtagtagttg tgccgttacc agtagcaggg gcgttaaacg agaatgatgt
     3121 ggagtttgct gaggcattgg taccattggt gttagcagta ccgaatgaaa atgcagattt
     3181 agatgttgta ttaccagtag cgtctgttgg cttacccaag acaggaatcg gcgttgagga
     3241 agcagaacca tcgaagaaag aagttggaga gtttgacttt tctttattgt gattgaactt
     3301 tgtaaaggaa aagccatttg atagcttctc cgtatttgct gccgctgtac ttgctgtcga
     3361 cttcatcgac tcgggagccc caaaactaaa agatggtttt gtgctagtgg ttgttgtatt
     3421 atttgttgtg gaaccgccaa aggtgaaaga tggcggtgta ggtcttttat ctgtctcatt
     3481 agcaggaggt ttagtgaaag aaaatgaggt gttagagcct ggtggttctt tagctgcatc
     3541 tgacttacca aatgaaaaca atggctttgg ttgtgaggtt tttgaagacg caaacgtaaa
     3601 ggagggcttt ttaggttccg aaacaacaga tgaatctttt tgagcaggtt cagtaaaaga
     3661 aaaagttggc ttgagagtct tatcatccgt cggtgcttga acatcaacag gcttgcccgg
     3721 aaacgaaaac gagggtttag ctgcttcgtt tgaaattgga ctactcttac gttcctcctc
     3781 tgacttagag aaagagaatg taggtttcgc acttccctca gagatcttat tttcacttgt
     3841 tgactgccca aaagtaaaag taggcttctt gactattgtg gcaggtgtct cagatggttt
     3901 ggtgtgtgtt tctttcgcgg tggcggcttt accaaaggta aattgtgcag aggagtcaat
     3961 attgcttgtt acatcagctt tttttccgaa tgtaaataat ggtgtacctt cagcttgctt
     4021 atcacttgca ccaaagacaa agcttggttt ccctgatgcg tccttttctg actctccctt
     4081 tttggtctcc ttttgatcac cggtcttgcc gaaatcgaat aaaggcttgg tgtttgtatc
     4141 ctcgctaaca ggtaaacgcc tttttctttt gggctcattt tcatcatcac cagcgtagtc
     4201 tgggacgtcg tatgggtatt catcaccatt ctcttcttgt ttaccaaaag aaaatattgg
     4261 agcagttgat tttggaggcg cgtctgattc tgtatgattt tcactttttt cggatgtctt
     4321 tccaaattta aaaggttgac tggcagaagt aacggtatct gatttaccac caaaattgaa
     4381 taaagttgtg gaagggacag tattgtcgac agccttagtt ttattagcct tttggctaaa
     4441 attgaatgat aaggtaggcg cctcggcagt tttcgttgac ttatcggttt ttcccatttc
     4501 tacactcgat ttaaagactg cacctgcaga agaagttgcc ttaggagaag ttttcttaga
     4561 tggagtctca ttgtccttga taaagtcaaa acctacggtg ggcaaaacaa tactttcttt
     4621 gtcctttttg ggctcaatat ttttctttaa cgtaggcgta ccagagcgct ctgaattggg
     4681 aacaaagctc tcctgaatag gggcagtttt tgcaactgtg gaagctggtc cttttaaaag
     4741 tagatttttt tgaggattcg ataacctatt agagttgatg tctgcacgta ggtcttcaat
     4801 ttcgcttgtc aggttagggc ctgtagccag attgccattt gaaatactac tcttaatatt
     4861 atttctattc tcgcttgtct tctgatcacc gccagcgtta ccttccttat ccttgttatc
     4921 ctttttttgt atagcgtcat attctgacaa atcatattca aaatttgctg accacacggt
     4981 cccctttgac tgactatgaa acctttttct attggatcta tttttcaatg atttgagaat
     5041 gggtagtcca acattggtgt cttcaccgct ttttccggcc aactgcctag tgcaagaacc
     5101 gtttttaata ggggaaggag tagatgatgt gcataggtac gatcctccct catcgctttt
     5161 actttgagag cccaatataa ccgacgatgt aatagatgga aattcagttg attgaattaa
     5221 tccaagctca cgcatatttc tcaccctctg cttctccctt aataacctca gtctttgaat
     5281 gggcaaaatt ggcaaaagcg gcggtctctc agtgttttcg gttccatata ttataattgg
     5341 cgcattattg ttgttctgag ttaagctgtc gctatgctgt gatgtaccgg acaccctctt
     5401 tctcttatta acatgcagtg tgtcttcaac atctgattcc tccaaatgat tcgcgtatga
     5461 gaggtttgaa ctgaaaactt tcttgctcga tggccgtttt ttattggggt ttgtgaagaa
     5521 tgattttaaa gtggaagaaa acgatctctt ttcgacacgt ggagaagaca tcacagaaga
     5581 agtgtttgaa gacatgaatg actaaaaatt gtcgctcact ctctgtccct ataacccttt
     5641 cgaggctaat atcctatcgt atttgcaccg ctacgtagtg tccttattga gttcctcatc
     5701 acttattttc tttaagtgtt tcttgacatt acgaaatttc gtcaaagaaa aaaattaaaa
     5761 tgaaaaagca tttcaatgtc acataatacg aaccattgat cacgtgcaac gacaaaccct
     5821 aaatataaaa actagggcgt aaaaaccggg gcttgaaaat tagggcataa aataggcttt
     5881 gcatacacgt gacttatatt tggtgtcggc gttttcttta cgcggtgtag tgtaaatctc
     5941 ttgtcgtaca agtggatata cgcactgtat acctccagta acaccaaaaa aaaaaccgtg
     6001 gttgtcccat gtaaacgagt accgcacacg taggccaaag cactccagag agacttcgtg
     6061 tcaaaggtct ataataggtg gtgccttctt gcttcttttt tgcagattct tagtataata
     6121 cgctagacta ttgtactttc taattttaag agatatcaag cttatcgata ccgtcgacct
     6181 cgaggggggg cccggtacca gcttttgttc cctttagtga gggttaattt cgagcttggc
     6241 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6301 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6361 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6421 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6481 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6541 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6601 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6661 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6721 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6781 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6841 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6901 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6961 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7021 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7081 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7141 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7201 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7261 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7321 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7381 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7441 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7501 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7561 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7621 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7681 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7741 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7801 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7861 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7921 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7981 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8041 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8101 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8161 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8221 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8281 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8341 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8401 tgggtccttt tcatcacgtg ctataaaaat aattataatt taaatttttt aatataaata
     8461 tataaattaa aaatagaaag taaaaaaaga aattaaagaa aaaatagttt ttgttttccg
     8521 aagatgtaaa agactctagg gggatcgcca acaaatacta ccttttatct tgctcttcct
     8581 gctctcaggt attaatgccg aattgtttca tcttgtctgt gtagaagacc acacacgaaa
     8641 atcctgtgat tttacatttt acttatcgtt aatcgaatgt atatctattt aatctgcttt
     8701 tcttgtctaa taaatatata tgtaaagtac gctttttgtt gaaatttttt aaacctttgt
     8761 ttattttttt ttcttcattc cgtaactctt ctaccttctt tatttacttt ctaaaatcca
     8821 aatacaaaac ataaaaataa ataaacacag agtaaattcc caaattattc catcattaaa
     8881 agatacgagg cgcgtgtaag ttacaggcaa gcgatccgtc taagaaacca ttattatcat
     8941 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtc
//
