LOCUS       sequence-367560-        7184 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of full length NUP2 as fusion with GST.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7184)
  AUTHORS   Ken Belanger Lab
  TITLE     Belanger lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7184)
  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: 
            "Unpublished"
COMMENT     Sequence Label: LDB361
FEATURES             Location/Qualifiers
     source          1..7184
                     /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             complement(2193..2204)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3234..3256)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3401..3419)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3487..3591
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3592..3660,3661..4452)
                     /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(3810..3829)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4623..5211
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5112..5131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5365..5382
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5455..5532
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5533..6615
                     /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(5552..5571)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6664..6669,6670..6687,6688..6694)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6702..6718
                     /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     6726..6742
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6726..6742
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6738..7184,1..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(6758..6775)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6758..6774)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6767..6789)
                     /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 ggatccgcca
      961 aaagagttgc cgatgcgcga atacagagag aaacgtacga ttctaacgag tctgacgatg
     1021 acgtgactcc ctccactaag gttgcgtcat ctgctgtgat gaatagaaga aaaattgcca
     1081 tgccaaagcg caggatggcg ttcaaacctt ttggttctgc aaaatcggat gaaaccaagc
     1141 aggctagttc ctttagcttc ctgaaccggg cggacggcac tggagaagct caggttgata
     1201 atagccctac cacagaaagc aattccagac taaaagcatt gaacctccag ttcaaggcta
     1261 aggttgatga cttagttcta ggcaagccgt tagcggactt gaggcccctt ttcaccaggt
     1321 acgaattata cataaagaat atcttagaag ctcccgtgaa atctatcgag aatccaacgc
     1381 agacaaaggg aaatgatgct aaacctgcca aagtagaaga tgtccaaaaa agttccgatt
     1441 cttcatctga agatgaggtt aaggtggagg gacccaagtt cacaatagat gctaaaccgc
     1501 ctatttcaga ttccgttttc tcatttggcc caaaaaaaga aaatcgcaag aaagatgaaa
     1561 gtgatagcga aaacgatata gaaatcaagg gccctgaatt taaattttct ggaactgtat
     1621 caagtgatgt atttaagctg aatccaagca ccgataaaaa tgaaaagaaa accgagacta
     1681 atgctaaacc attttcattt tcttcggcca cttcaactac tgaacaaacg aagagtaaaa
     1741 atcccctttc attgacagaa gctaccaaga ccaatgtgga caacaacagt aaagccgagg
     1801 cttccttcac ttttggaaca aaacatgctg cggattctca aaataataaa ccatcttttg
     1861 tatttggtca agcagctgca aaaccatcgc tagaaaagag ctcattcacg tttggttcaa
     1921 caacaattga aaaaaaaaat gacgaaagct caacctctaa ctcaaaacct gaaaagtcta
     1981 gtgatagcaa tgattcaaac ccatcttttt ccttttccat acccagtaag aatacacctg
     2041 atgcatctaa gccatctttt agttttgggg tcccaaactc ttccaaaaac gaaacttcaa
     2101 aaccggtatt ttcgtttggt gcagcaacac catcggccaa agaagctagt caggaagatg
     2161 acaacaacaa cgttgaaaaa ccttcctcta agcctgcctt caatttaata tctaacgctg
     2221 gtaccgagaa agagaaggaa agtaaaaagg actcaaagcc agctttttca tttggcatat
     2281 caaacggaag tgaaagcaaa gactctgaca aaccctcttt accctctgcg gttgatggtg
     2341 aaaatgacaa gaaagaagca acaaaacctg ctttttcgtt tggaataaat acaaatacta
     2401 ctaaaaccgc ggatactaaa gctccaactt ttacatttgg ctcctctgca ctcgctgaca
     2461 ataaagagga tgttaagaaa cctttttctt tcggtacctc ccagcctaat aatactccat
     2521 ccttctcatt cggaaaaaca acagcaaact tgcctgctaa ttcttcaaca tctcctgctc
     2581 cctctatacc atcgacgggg ttcaaatttt ctttgccatt tgaacaaaaa ggtagtcaaa
     2641 caactacaaa tgatagcaag gaagaatcaa caacagaagc aactggaaat gagtcgcaag
     2701 atgcaaccaa agtagatgct accccagaag aatcaaagcc aataaacttg caaaacggtg
     2761 aggaagacga agtggcttta ttttcgcaaa aagcaaaatt aatgacattc aatgctgaaa
     2821 ccaaatcgta cgattcaaga ggcgtaggcg aaatgaagct tttgaagaaa aaggacgatc
     2881 cttctaaagt gcgcctactt tgtaggtctg acggtatggg taatgtatta ctaaatgcaa
     2941 ctgttgtaga ctccttcaaa tatgagcctt tagctcccgg aaatgataat ctcattaaag
     3001 ctcctactgt tgcggctgat gggaaacttg taacttatat cgtcaagttt aagcagaagg
     3061 aagaaggccg ctcatttacg aaagctattg aagatgctaa aaaagaaatg aaataagagt
     3121 aaatacagtt aacaatttta aatagaatag aaccctcata ttttgtagct caagggaatt
     3181 catcgtgact gactgacgat ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     3241 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     3301 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
     3361 cgtagcgata gcggagtgta taattcttga agacgaaagg gcctcgtgat acgcctattt
     3421 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3481 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3541 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3601 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3661 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3721 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3781 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     3841 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3901 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3961 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4021 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4081 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     4141 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4201 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4261 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4321 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4381 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4441 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4501 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4561 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4621 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4681 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4741 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     4801 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4861 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4921 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4981 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5041 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5101 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5161 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5221 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5281 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     5341 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     5401 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcataaa ttccgacacc
     5461 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
     5521 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
     5581 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
     5641 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
     5701 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
     5761 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
     5821 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
     5881 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
     5941 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
     6001 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
     6061 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
     6121 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
     6181 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
     6241 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
     6301 attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc
     6361 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
     6421 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     6481 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
     6541 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     6601 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcacccca
     6661 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     6721 tcacacagga aacagctatg accatgatta cggattcact ggccgtcgtt ttacaacgtc
     6781 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     6841 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     6901 tgaatggcga atggcgcttt gcctggtttc cggcaccaga agcggtgccg gaaagctggc
     6961 tggagtgcga tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg
     7021 gttacgatgc gcccatctac accaacgtga cctatcccat tacggtcaat ccgccgtttg
     7081 ttcccacgga gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc
     7141 tacaggaagg ccagacgcga attatttttg atggcgttgg aatt
//
