LOCUS       sequence-367551-        6108 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of N-terminus of NUP1 nucleoporin as GST-Nup1-N
            fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6108)
  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 6108)
  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: LDB203
FEATURES             Location/Qualifiers
     source          1..6108
                     /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(2158..2180)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2325..2343)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2411..2515
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2516..2584,2585..3376)
                     /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(2734..2753)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3547..4135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4036..4055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4289..4306
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4379..4456
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4457..5539
                     /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(4476..4495)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5588..5593,5594..5611,5612..5618)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5626..5642
                     /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     5650..5666
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5650..5666
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5662..6108,1..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(5682..5699)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5682..5698)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5691..5713)
                     /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 ggatccactt
      961 cttctgtgat gtcttctcca cgtgtcgaaa agagatcgtt ttcttccact ttaaaatcat
     1021 tcttcacaaa ccccaataaa aaacggccat cgagcaagaa agttttcagt tcaaacctct
     1081 catacgcgaa tcatttggag gaatcagatg ttgaagacac actgcatgtt aataagagaa
     1141 agagggtgtc cggtacatca cagcatagcg acagcttaac tcagaacaac aataatgcgc
     1201 caattataat atatggaacc gaaaacactg agagaccgcc gcttttgcca attttgccca
     1261 ttcaaagact gaggttatta agggagaagc agagggtgag aaatatgcgt gagcttggat
     1321 taattcaatc aactgaattt ccatctatta catcgtcggt tatattgggc tctcaaagta
     1381 aaagcgatga gggaggatcg tacctatgca catcatctac tccttcccct attaaaaacg
     1441 gttcttgcac taggcagttg gccggaaaaa gcggtgaaga caccaatgtt ggactaccca
     1501 ttctcaaatc attgaaaaat agatccaata gaaaaaggtt tcatggtcag tcaaagggga
     1561 ccgtatggtc agcaaatttt gaatatgatt tgtcagaata tgacgctata caaaaaaagg
     1621 ataacaagga taaggaaggt aacgctggcg gtgatcagaa gacaagcgag aatagaaata
     1681 atattaagag tagtatttca aatggcaatc tggctacagg ccctaacctg acaagcgaaa
     1741 ttgaagacct acgtgcagac atcaactcta ataggttatc gaatcctcaa aaaaatctac
     1801 ttttaaaagg accagcttcc acagttgcaa aaactgcccc tattcaggag agctttgttc
     1861 ccaattcaga gcgctctggt acgcctacgt taaagaaaaa tattgagccc aaaaaggaca
     1921 aagaaagtat tgttttgccc accgtaggtt ttgactttat caaggacaat gagactccat
     1981 ctaagaaaac ttctcctaag gcagcttctt ctgcaggtgc agtctttaaa tcgagtgtag
     2041 aaatgggaaa aaccgataag tcaacgaaaa ctgccgaggc gcctacctta tcattctagg
     2101 aattcatcgt gactgactga cgatctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     2161 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     2221 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     2281 gtcacgtagc gatagcggag tgtataattc ttgaagacga aagggcctcg tgatacgcct
     2341 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     2401 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2461 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2521 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2581 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2641 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2701 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     2761 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2821 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2881 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     2941 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3001 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     3061 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3121 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3181 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3241 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3301 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3361 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3421 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3481 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3541 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3601 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3661 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     3721 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3781 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3841 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3901 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3961 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4021 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4081 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4141 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4201 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4261 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4321 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca taaattccga
     4381 caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
     4441 attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt
     4501 ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg
     4561 ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca
     4621 actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc
     4681 gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt
     4741 ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct
     4801 cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc
     4861 tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc
     4921 catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt
     4981 cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg
     5041 tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga
     5101 acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga
     5161 gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg
     5221 cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga
     5281 taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct
     5341 gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg
     5401 caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca
     5461 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     5521 actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac
     5581 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     5641 aatttcacac aggaaacagc tatgaccatg attacggatt cactggccgt cgttttacaa
     5701 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     5761 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     5821 agcctgaatg gcgaatggcg ctttgcctgg tttccggcac cagaagcggt gccggaaagc
     5881 tggctggagt gcgatcttcc tgaggccgat actgtcgtcg tcccctcaaa ctggcagatg
     5941 cacggttacg atgcgcccat ctacaccaac gtgacctatc ccattacggt caatccgccg
     6001 tttgttccca cggagaatcc gacgggttgt tactcgctca catttaatgt tgatgaaagc
     6061 tggctacagg aaggccagac gcgaattatt tttgatggcg ttggaatt
//
