LOCUS       sequence-364127-        5288 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of C-terminus of NUP1 nucleoporin as GST-Nup1-C
            fusion truncated after the last FXFG repeat.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5288)
  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 5288)
  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: KBB101
FEATURES             Location/Qualifiers
     source          1..5288
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(137..142,143..160,161..167)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    175..191
                     /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     199..215
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     199..215
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             211..735
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVISLSTARCTNASGVRQ
                     PSEAVVWLCRS"
     primer_bind     complement(231..248)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(231..247)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(240..262)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(837..842,843..858,859..865)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    873..889
                     /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             912..1565
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1523..1545
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1572..1589
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2162..2180)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2248..2352
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2353..2421,2422..3213)
                     /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(2571..2590)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3384..3972
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3873..3892
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4126..4143
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4216..4293
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(4294..5288,1..88)
                     /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(4313..4332)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
       61 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagctcactc
      121 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
      181 gcggataaca atttcacaca ggaaacagct atgaccatga ttacggattc actggccgtc
      241 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
      301 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
      361 cagttgcgca gcctgaatgg cgaatggcgc tttgcctggt ttccggcacc agaagcggtg
      421 ccggaaagct ggctggagtg cgatcttcct gaggccgata ctgtcgtcgt cccctcaaac
      481 tggcagatgc acggttacga tgcgcccatc tacaccaacg tgacctatcc cattacggtc
      541 aatccgccgt ttgttcccac ggagaatccg acgggttgtt actcgctcac atttaatgtt
      601 gatgaaagct ggctacagga aggccagacg cgaattattt ttgatggcgt aattagctta
      661 tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc tgtggtatgg
      721 ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca aggcgcactc ccgttctgga
      781 taatgttttt tgcgccgaca tcataacggt tctggcaaat attctgaaat gagctgttga
      841 caattaatca tcggctcgta taatgtgtgg aattgtgagc ggataacaat ttcacacagg
      901 aaacagtatt catgtcccct atactaggtt attggaaaat taagggcctt gtgcaaccca
      961 ctcgacttct tttggaatat cttgaagaaa aatatgaaga gcatttgtat gagcgcgatg
     1021 aaggtgataa atggcgaaac aaaaagtttg aattgggttt ggagtttccc aatcttcctt
     1081 attatattga tggtgatgtt aaattaacac agtctatggc catcatacgt tatatagctg
     1141 acaagcacaa catgttgggt ggttgtccaa aagagcgtgc agagatttca atgcttgaag
     1201 gagcggtttt ggatattaga tacggtgttt cgagaattgc atatagtaaa gactttgaaa
     1261 ctctcaaagt tgattttctt agcaagctac ctgaaatgct gaaaatgttc gaagatcgtt
     1321 tatgtcataa aacatattta aatggtgatc atgtaaccca tcctgacttc atgttgtatg
     1381 acgctcttga tgttgtttta tacatggacc caatgtgcct ggatgcgttc ccaaaattag
     1441 tttgttttaa aaaacgtatt gaagctatcc cacaaattga taagtacttg aaatccagca
     1501 agtatatagc atggcctttg cagggctggc aagccacgtt tggtggtggc gaccatcctc
     1561 caaaatcgga tctggttccg cgtggatctc gtcgtgcatc tgttggatcc tcaacgccga
     1621 ttcctgtctt gggtaagcca acagacgcta ctggtaatac aacatctaaa tctgcatttt
     1681 cattcggtac tgctaacacc aatggtacca atgcctcagc aaactccaca tcattctcgt
     1741 ttaacgcccc tgctactggt aacggcacaa ctactacttc caatacctca ggaaccaata
     1801 tagcgggtac atttaacgta ggaaaaccgg atcaaagtat cgctagtggc aatacgaatg
     1861 gagcgggctc ggcatttggc ttttcgagct caggaacagc agcaactggt gcagcttcta
     1921 atcaatcttc atttaatttt ggaaacaatg gtgcaggggg tctcaatcct tttacatcag
     1981 caacttcgtc aactaatgct aatgctggtt tattcaataa acctccttcc acgaatgcac
     2041 aaaatgtcaa tgttccctct gcttttaatt ttacgggaaa taattcaacg cctggtggcg
     2101 gctctgtatt taatatgaac ggcaacacta atgctaatac ggtgtttgcc aagacgaaag
     2161 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     2221 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2281 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2341 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     2401 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     2461 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     2521 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     2581 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     2641 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     2701 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     2761 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     2821 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     2881 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     2941 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3001 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3061 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3121 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     3181 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     3241 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3301 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3361 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3421 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3481 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     3541 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3601 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3661 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3721 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3781 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3841 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3901 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3961 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4021 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4081 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     4141 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4201 caccgcataa attccgacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag
     4261 cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc
     4321 gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac
     4381 gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc
     4441 aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc
     4501 agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa
     4561 ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg
     4621 gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat
     4681 gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat
     4741 gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg
     4801 ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta
     4861 agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa
     4921 attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc
     4981 atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg
     5041 gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg
     5101 gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc
     5161 aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag
     5221 ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc
     5281 ctggcgcc
//
