LOCUS       sequence-367570-        5898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of Nup1 C-terminal residues 777-1076 as GST 
            fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5898)
  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 5898)
  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: LDB221
FEATURES             Location/Qualifiers
     source          1..5898
                     /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"
     primer_bind     complement(1948..1970)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2115..2133)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2201..2305
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2306..2374,2375..3166)
                     /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(2524..2543)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3337..3925
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3826..3845
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4079..4096
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4169..4246
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4247..5329
                     /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(4266..4285)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5378..5383,5384..5401,5402..5408)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5416..5432
                     /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     5440..5456
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5440..5456
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5452..5898,1..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(5472..5489)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5472..5488)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5481..5503)
                     /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 ggatcctcaa
      961 cgccgattcc tgtcttgggt aagccaacag acgctactgg taatacaaca tctaaatctg
     1021 cattttcatt cggtactgct aacaccaatg gtaccaatgc ctcagcaaac tccacatcat
     1081 tctcgtttaa cgcccctgct actggtaacg gcacaactac tacttccaat acctcaggaa
     1141 ccaatatagc gggtacattt aacgtaggaa aaccggatca aagtatcgct agtggcaata
     1201 cgaatggagc gggctcggca tttggctttt cgagctcagg aacagcagca actggtgcag
     1261 cttctaatca atcttcattt aattttggaa acaatggtgc agggggtctc aatcctttta
     1321 catcagcaac ttcgtcaact aatgctaatg ctggtttatt caataaacct ccttccacga
     1381 atgcacaaaa tgtcaatgtt ccctctgctt ttaattttac gggaaataat tcaacgcctg
     1441 gtggcggctc tgtatttaat atgaacggca acactaatgc taatacggtg tttgccggct
     1501 ctaataacca accacatcaa tcgcaaaccc catctttcaa tacaaacagc tcattcacgc
     1561 catcaacagt tcctaatatt aattttagcg gattgaatgg cggaattact aataccgcga
     1621 ccaatgcatt aaggccaagt gatatatttg gtgcgaatgc tgcctctggt tccaattcaa
     1681 acgtaacaaa tccatcatcc atttttgggg gggcaggtgg tgtgccgaca acttcttttg
     1741 ggcagccgca gtcagcccct aatcagatgg ggatgggaac aaataatggc atgagcatgg
     1801 gcggtggtgt tatggcgaac agaaagattg caagaatgag gcactctaaa aggtaaatat
     1861 cggttatgta attaggtatt gtgttgtagg aattcatcgt gactgactga cgatctgcct
     1921 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     1981 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     2041 tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtataattc
     2101 ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     2161 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2221 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     2281 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     2341 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     2401 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     2461 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     2521 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     2581 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     2641 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     2701 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     2761 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     2821 aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt
     2881 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     2941 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     3001 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     3061 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     3121 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     3181 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     3241 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3301 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3361 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3421 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3481 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3541 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3601 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3661 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3721 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3781 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3841 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3901 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3961 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     4021 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     4081 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     4141 gtgcggtatt tcacaccgca taaattccga caccatcgaa tggcgcaaaa cctttcgcgg
     4201 tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac
     4261 gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa
     4321 ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct
     4381 gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg
     4441 cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc
     4501 tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga
     4561 agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa
     4621 ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc
     4681 gttatttctt gatgtctctg accagacacc catcaacagt attattttct cccatgaaga
     4741 cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt
     4801 agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct
     4861 cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg
     4921 ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc
     4981 caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg
     5041 tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc
     5101 gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct
     5161 gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa
     5221 aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     5281 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     5341 ttaatgtaag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     5401 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     5461 attacggatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     5521 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     5581 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg ctttgcctgg
     5641 tttccggcac cagaagcggt gccggaaagc tggctggagt gcgatcttcc tgaggccgat
     5701 actgtcgtcg tcccctcaaa ctggcagatg cacggttacg atgcgcccat ctacaccaac
     5761 gtgacctatc ccattacggt caatccgccg tttgttccca cggagaatcc gacgggttgt
     5821 tactcgctca catttaatgt tgatgaaagc tggctacagg aaggccagac gcgaattatt
     5881 tttgatggcg ttggaatt
//
