LOCUS       sequence-367180-        5235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of C-terminus of NUP1 nucleoporin as GST-Nup1-C
            fusion including C-terminal half of Nup1 C-terminal FG domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5235)
  AUTHORS   Belanger KD
  TITLE     Using affinity chromatography to investigate novel protein-protein 
            interactions in an undergraduate cell and molecular biology lab 
            course.
  JOURNAL   CBE Life Sci Educ. 2009 Fall;8(3):214-25. doi: 
            10.1187/cbe.09-03-0019.
  PUBMED    19723816
REFERENCE   2  (bases 1 to 5235)
  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: "10.1187/cbe"; 
            journalName: "CBE Life Sci Educ"; date: "2009-?"; volume: "8"; 
            issue: "3"; pages: "214-25"
COMMENT     Sequence Label: KBB106
FEATURES             Location/Qualifiers
     source          1..5235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(523..528,529..546,547..553)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    561..577
                     /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     585..601
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     585..601
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             597..1124
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(617..634)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(617..633)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(626..648)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1226..1231,1232..1247,1248..1254)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1262..1278
                     /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             1301..1954
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1912..1934
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1961..1978
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2328..2350)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2495..2513)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2581..2685
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2686..2754,2755..3546)
                     /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(2904..2923)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3717..4305
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4206..4225
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4459..4476
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4549..4626
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(4627..5235,1..474)
                     /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(4646..4665)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
       61 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
      121 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
      181 gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc
      241 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
      301 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg
      361 gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc
      421 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
      481 cgcaattaat gtaagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
      541 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
      601 ccatgattac ggattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
      661 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
      721 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg
      781 cctggtttcc ggcaccagaa gcggtgccgg aaagctggct ggagtgcgat cttcctgagg
      841 ccgatactgt cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg cccatctaca
      901 ccaacgtgac ctatcccatt acggtcaatc cgccgtttgt tcccacggag aatccgacgg
      961 gttgttactc gctcacattt aatgttgatg aaagctggct acaggaaggc cagacgcgaa
     1021 ttatttttga tggcgttgga attagcttat cgactgcacg gtgcaccaat gcttctggcg
     1081 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
     1141 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
     1201 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga
     1261 attgtgagcg gataacaatt tcacacagga aacagtattc atgtccccta tactaggtta
     1321 ttggaaaatt aagggccttg tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa
     1381 atatgaagag catttgtatg agcgcgatga aggtgataaa tggcgaaaca aaaagtttga
     1441 attgggtttg gagtttccca atcttcctta ttatattgat ggtgatgtta aattaacaca
     1501 gtctatggcc atcatacgtt atatagctga caagcacaac atgttgggtg gttgtccaaa
     1561 agagcgtgca gagatttcaa tgcttgaagg agcggttttg gatattagat acggtgtttc
     1621 gagaattgca tatagtaaag actttgaaac tctcaaagtt gattttctta gcaagctacc
     1681 tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa acatatttaa atggtgatca
     1741 tgtaacccat cctgacttca tgttgtatga cgctcttgat gttgttttat acatggaccc
     1801 aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa aaacgtattg aagctatccc
     1861 acaaattgat aagtacttga aatccagcaa gtatatagca tggcctttgc agggctggca
     1921 agccacgttt ggtggtggcg accatcctcc aaaatcggat ctggttccgc gtggatctcg
     1981 tcgtgcatct gttggatccc cgggaatttt aaggccaagt gatatatttg gtgcgaatgc
     2041 tgcctctggt tccaattcaa acgtaacaaa tccatcatcc atttttgggg gggcaggtgg
     2101 tgtgccgaca acttcttttg ggcagccgca gtcagcccct aatcagatgg ggatgggaac
     2161 aaataatggc atgagcatgg gcggtggtgt tatggcgaac agaaagattg caagaatgag
     2221 gcactctaaa aggtaaatat cggttatgta attaggtatt gtgttgtagg aattcatcgt
     2281 gactgactga cgatctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     2341 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     2401 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
     2461 gatagcggag tgtataattc ttgaagacga aagggcctcg tgatacgcct atttttatag
     2521 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     2581 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2641 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     2701 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     2761 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     2821 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     2881 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     2941 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3001 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3061 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3121 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3181 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     3241 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3301 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3361 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     3421 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     3481 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3541 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3601 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3661 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3721 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3781 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3841 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     3901 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3961 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4021 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4081 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4141 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4201 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4261 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4321 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4381 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4441 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     4501 tattttctcc ttacgcatct gtgcggtatt tcacaccgca taaattccga caccatcgaa
     4561 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
     4621 gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag
     4681 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg
     4741 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc
     4801 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa
     4861 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg
     4921 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc
     4981 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct
     5041 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt
     5101 attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt
     5161 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg
     5221 gctggctggc ataaa
//
