LOCUS       sequence-368132-        7567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminus and FXFG repeats of NUP1 
            nucleoporin as GST-Nup1-N+Rep fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7567)
  AUTHORS   Ken Belanger Lab
  TITLE     Belanger lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7567)
  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            "Unpublished"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: KBB258
FEATURES             Location/Qualifiers
     source          1..7567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(493..498,499..516,517..523)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    531..547
                     /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     555..571
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     555..571
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             567..1094
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(587..604)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(587..603)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(596..618)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1196..1201,1202..1217,1218..1224)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1232..1248
                     /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             1271..1924
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1882..1904
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1931..1948
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(4630..4652)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4797..4815)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4883..4987
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4988..5056,5057..5848)
                     /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(5206..5225)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6019..6607
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6508..6527
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6761..6778
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        6851..6928
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(6929..7567,1..444)
                     /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(6948..6967)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
       61 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
      121 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac
      181 gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt
      241 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
      301 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat
      361 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      421 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta
      481 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      541 ataacaattt cacacaggaa acagctatga ccatgattac ggattcactg gccgtcgttt
      601 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
      661 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
      721 tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa gcggtgccgg
      781 aaagctggct ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc tcaaactggc
      841 agatgcacgg ttacgatgcg cccatctaca ccaacgtgac ctatcccatt acggtcaatc
      901 cgccgtttgt tcccacggag aatccgacgg gttgttactc gctcacattt aatgttgatg
      961 aaagctggct acaggaaggc cagacgcgaa ttatttttga tggcgttgga attagcttat
     1021 cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc atcggaagct gtggtatggc
     1081 tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa ggcgcactcc cgttctggat
     1141 aatgtttttt gcgccgacat cataacggtt ctggcaaata ttctgaaatg agctgttgac
     1201 aattaatcat cggctcgtat aatgtgtgga attgtgagcg gataacaatt tcacacagga
     1261 aacagtattc atgtccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac
     1321 tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga
     1381 aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca atcttcctta
     1441 ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga
     1501 caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg
     1561 agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac
     1621 tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt
     1681 atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga
     1741 cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt
     1801 ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa
     1861 gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc
     1921 aaaatcggat ctggttccgc gtggatctcg tcgtgcatct gttggatccg aatatgattt
     1981 gtcagaatat gacgctatac aaaaaaagga taacaaggat aaggaaggta acgctggcgg
     2041 tgatcagaag acaagcgaga atagaaataa tattaagagt agtatttcaa atggcaatct
     2101 ggctacaggc cctaacctga caagcgaaat tgaagaccta cgtgcagaca tcaactctaa
     2161 taggttatcg aatcctcaaa aaaatctact tttaaaagga ccagcttcca cagttgcaaa
     2221 aactgcccct attcaggaga gctttgttcc caattcagag cgctctggta cgcctacgtt
     2281 aaagaaaaat attgagccca aaaaggacaa agaaagtatt gttttgccca ccgtaggttt
     2341 tgactttatc aaggacaatg agactccatc taagaaaact tctcctaagg caacttcttc
     2401 tgcaggtgca gtctttaaat cgagtgtaga aatgggaaaa accgataagt caacgaaaac
     2461 tgccgaggcg cctaccttat cattcaattt tagccaaaag gctaataaaa ctaaggctgt
     2521 cgacaatact gtcccttcca caactttatt caattttggt ggtaaatcag ataccgttac
     2581 ttctgccagt caacctttta aatttggaaa gacatccgaa aaaagtgaaa atcatacaga
     2641 atcagacgcg cctccaaaat caactgctcc aatattttct tttggtaaac aagaagagaa
     2701 tggtgatgaa ggtgatgatg aaaatgagcc caaaagaaaa aggcgtttac ctgttagcga
     2761 ggatacaaac accaagcctt tattcgattt cggcaagacc ggtgatcaaa aggagaccaa
     2821 aaagggagag tcagaaaagg acgcatcagg gaaaccaagc tttgtctttg gtgcaagtga
     2881 taagcaagct gaaggtacac cattatttac attcggaaaa aaagctgatg taacaagcaa
     2941 tattgactcc tctgcacaat ttacctttgg taaagccgcc accgcgaaag aaacacacac
     3001 caaaccatct gagacacctg ccacaatagt caagaagcct acttttactt ttgggcagtc
     3061 aacaagtgaa aataagatct ctgagggaag tgcgaaacct acattctctt tctctaagtc
     3121 agaggaggaa cgtaagagta gtccaatttc aaacgaagca gctaaaccct cgttttcgtt
     3181 tccgggcaag cctgttgatg ttcaagcacc gacggatgat aagactctca agccaacttt
     3241 ttcttttact gaacctgctc aaaaagattc atctgttgtt tcggaaccta aaaagccctc
     3301 ctttacgttt gcgtcttcaa aaacctcaca accaaagcca ttgttttcat ttggtaagtc
     3361 agatgcagct aaagaaccac caggctctaa cacctcattt tctttcacta aacctcctgc
     3421 taatgagaca gataaaagac ctacaccgcc atctttcacc tttggcggtt ccacaacaaa
     3481 taatacaaca accactagca caaaaccatc ttttagtttt ggggctcccg agtcgatgaa
     3541 gtcgacagca agtacagcgg cagcaaatac ggagaagcta tcaaatggct tttcctttac
     3601 aaagttcaat cacaataaag aaaagtcaaa ctctccaact tctttcttcg atggttctgc
     3661 ttcctcaacg ccgattcctg tcttgggtaa gccaacagac gctactggta atacaacatc
     3721 taaatctgca ttttcattcg gtactgctaa caccaatggt accaatgcct cagcaaactc
     3781 cacatcattc tcgtttaacg cccctgctac tggtaacggc acaactacta cttccaatac
     3841 ctcaggaacc aatatagcgg gtacatttaa cgtaggaaaa ccggatcaaa gtatcgctag
     3901 tggcaatacg aatggagcgg gctcggcatt tggcttttcg agctcaggaa cagcagcaac
     3961 tggtgcagct tctaatcaat cttcatttaa ttttggaaac aatggtgcag ggggtctcaa
     4021 tccttttaca tcagcaactt cgtcaactaa tgctaatgct ggtttattca ataaacctcc
     4081 ttccacgaat gcacaaaatg tcaatgttcc ctctgctttt aattttacgg gaaataattc
     4141 aacgcctggt ggcggctctg tatttaatat gaacggcaac actaatgcta atacggtgtt
     4201 tgccggctct aataaccaac cacatcaatc gcaaacccca tctttcaata caaacagctc
     4261 attcacgcca tcaacagttc ctaatattaa ttttagcgga ttgaatggcg gaattactaa
     4321 taccgcgacc aatgcattaa ggccaagtga tatatttggt gcgaatgctg cctctggttc
     4381 caattcaaac gtaacaaatc catcatccat ttttgggggg gcaggtggtg tgccgacaac
     4441 ttcttttggg cagccgcagt cagcccctaa tcagatgggg atgggaacaa ataatggcat
     4501 gagcatgggc ggtggtgtta tggcgaacag aaagattgca agaatgaggc actctaaaag
     4561 gtaaatatcg ggaattcatc gtgactgact gacgatctgc ctcgcgcgtt tcggtgatga
     4621 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     4681 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
     4741 agccatgacc cagtcacgta gcgatagcgg agtgtataat tcttgaagac gaaagggcct
     4801 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     4861 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4921 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4981 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5041 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5101 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5161 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5221 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5281 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5341 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5401 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5461 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5521 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5581 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5641 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5701 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5761 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5821 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     5881 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5941 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6001 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6061 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6121 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     6181 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6241 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6301 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6361 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6421 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6481 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6541 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6601 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6661 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     6721 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     6781 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     6841 cataaattcc gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc
     6901 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
     6961 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
     7021 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
     7081 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
     7141 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
     7201 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
     7261 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
     7321 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
     7381 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
     7441 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
     7501 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca
     7561 gccgata
//
