LOCUS       sequence-367544-        6150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of FG-repeat domain of NUP1 nucleoporin as 
            GST-Nup1-Rep fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6150)
  AUTHORS   Belanger KD, Walter D, Henderson TA, Yelton AL, O'Brien TG, Belanger
            KG, Geier SJ, Fahrenkrog B
  TITLE     Nuclear localisation is crucial for the proapoptotic activity of the
            HtrA-like serine protease Nma111p.
  JOURNAL   J Cell Sci. 2009 Nov 1;122(Pt 21):3931-41. doi: 10.1242/jcs.056887. 
            Epub 2009 Oct 6.
  PUBMED    19808885
REFERENCE   2  (bases 1 to 6150)
  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: "J Cell 
            Sci."; date: "2009-11-1"; volume: "122(Pt 21):3931-41. doi"; pages: 
            " 10.1242/jcs.056887. Epub 2009 Oct 6"
COMMENT     Sequence Label: LDB204
FEATURES             Location/Qualifiers
     source          1..6150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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)."
     CDS             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..935
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1044..1070
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2200..2222)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2367..2385)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2453..2557
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2558..2626,2627..3418)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2776..2795)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3589..4177
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4078..4097
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4331..4348
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4421..4498
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4499..5581
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4518..4537)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5630..5635,5636..5653,5654..5660)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5668..5684
                     /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)."
     primer_bind     5692..5708
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5692..5708
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5704..6150,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5724..5741)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5724..5740)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5733..5755)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gttccgcgtg gatctcgtcg tgcatctgtt ggatccagtg
      961 aaaatcatac agaatcagac gcgcctccaa aatcaactgc tccaatattt tcttttggta
     1021 aacaagaaga gaatggtgat gaatacccat acgacgtccc agactacgct ggtgatgatg
     1081 aaaatgagcc caaaagaaaa aggcgtttac ctgttagcga ggatacaaac accaagcctt
     1141 tattcgattt cggcaagacc ggtgatcaaa aggagaccaa aaagggagag tcagaaaagg
     1201 acgcatcagg gaaaccaagc tttgtctttg gtgcaagtga taagcaagct gaaggtacac
     1261 cattatttac attcggaaaa aaagctgatg taacaagcaa tattgactcc tctgcacaat
     1321 ttacctttgg taaagccgcc accgcgaaag aaacacacac caaaccatct gagacacctg
     1381 ccacaatagt caagaagcct acttttactt ttgggcagtc aacaagtgaa aataagatct
     1441 ctgagggaag tgcgaaacct acattctctt tctctaagtc agaggaggaa cgtaagagta
     1501 gtccaatttc aaacgaagca gctaaaccct cgttttcgtt tccgggcaag cctgttgatg
     1561 ttcaagcacc gacggatgat aagactctca agccaacttt ttcttttact gaacctgctc
     1621 aaaaagattc atctgttgtt tcggaaccta aaaagccctc ctttacgttt gcgtcttcaa
     1681 aaacctcaca accaaagcca ttgttttcat ttggtaagtc agatgcagct aaagaaccac
     1741 caggctctaa cacctcattt tctttcacta aacctcctgc taatgagaca gataaaagac
     1801 ctacaccgcc atctttcacc tttggcggtt ccacaacaaa taatacaaca accactagca
     1861 caaaaccatc ttttagtttt ggggctcccg agtcgatgaa gtcgacagca agtacagcgg
     1921 cagcaaatac ggagaagcta tcaaatggct tttcctttac aaagttcaat cacaataaag
     1981 aaaagtcaaa ctctccaact tctttcttcg atggttctgc ttcctcaacg ccgattcctg
     2041 tcttgggtaa gccaacagac gctactggta atacaacatc taaatctgca ttttcattcg
     2101 gtactgctaa caccaatggt accaatgcct cagcaaacta ggaattcatc gtgactgact
     2161 gacgatctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     2221 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     2281 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
     2341 agtgtataat tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     2401 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     2461 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2521 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     2581 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     2641 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     2701 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     2761 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     2821 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     2881 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     2941 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     3001 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     3061 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     3121 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     3181 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     3241 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     3301 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     3361 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     3421 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     3481 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     3541 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3601 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3661 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3721 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3781 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3841 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3901 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3961 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4021 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4081 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4141 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4201 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     4261 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     4321 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     4381 ccttacgcat ctgtgcggta tttcacaccg cataaattcc gacaccatcg aatggcgcaa
     4441 aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt
     4501 gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc
     4561 ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc
     4621 gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc
     4681 gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc
     4741 ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg
     4801 aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg
     4861 gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac
     4921 taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt
     4981 ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca
     5041 aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg
     5101 gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag
     5161 tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc
     5221 gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg
     5281 gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg
     5341 ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt
     5401 ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt
     5461 ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc
     5521 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     5581 agcgcaacgc aattaatgta agttagctca ctcattaggc accccaggct ttacacttta
     5641 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     5701 gctatgacca tgattacgga ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     5761 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     5821 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     5881 cgctttgcct ggtttccggc accagaagcg gtgccggaaa gctggctgga gtgcgatctt
     5941 cctgaggccg atactgtcgt cgtcccctca aactggcaga tgcacggtta cgatgcgccc
     6001 atctacacca acgtgaccta tcccattacg gtcaatccgc cgtttgttcc cacggagaat
     6061 ccgacgggtt gttactcgct cacatttaat gttgatgaaa gctggctaca ggaaggccag
     6121 acgcgaatta tttttgatgg cgttggaatt
//
