LOCUS       sequence-367178-        5631 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of C-terminus of NUP1 nucleoporin as GST-Nup1-C
            fusion truncated after NsiI restriction site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5631)
  AUTHORS   Ken Belanger Lab
  TITLE     Belanger lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5631)
  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            "Unpublished"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: KBB109
FEATURES             Location/Qualifiers
     source          1..5631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     10..26
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10..26
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             22..549
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(42..59)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(42..58)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(51..73)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(651..656,657..672,673..679)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    687..703
                     /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             726..1379
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1337..1359
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1386..1403
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2149..2171)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2316..2334)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2402..2506
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2507..2575,2576..3367)
                     /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(2725..2744)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3538..4126
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4027..4046
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4280..4297
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4370..4447
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4448..5530
                     /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(4467..4486)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5579..5584,5585..5602,5603..5609)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    join(5617..5631,1..2)
                     /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)."
ORIGIN
        1 aatttcacac aggaaacagc tatgaccatg attacggatt cactggccgt cgttttacaa
       61 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
      121 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
      181 agcctgaatg gcgaatggcg ctttgcctgg tttccggcac cagaagcggt gccggaaagc
      241 tggctggagt gcgatcttcc tgaggccgat actgtcgtcg tcccctcaaa ctggcagatg
      301 cacggttacg atgcgcccat ctacaccaac gtgacctatc ccattacggt caatccgccg
      361 tttgttccca cggagaatcc gacgggttgt tactcgctca catttaatgt tgatgaaagc
      421 tggctacagg aaggccagac gcgaattatt tttgatggcg ttggaattag cttatcgact
      481 gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg aagctgtggt atggctgtgc
      541 aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc actcccgttc tggataatgt
      601 tttttgcgcc gacatcataa cggttctggc aaatattctg aaatgagctg ttgacaatta
      661 atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca caggaaacag
      721 tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa cccactcgac
      781 ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc gatgaaggtg
      841 ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt ccttattata
      901 ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata gctgacaagc
      961 acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt gaaggagcgg
     1021 ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt gaaactctca
     1081 aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat cgtttatgtc
     1141 ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg tatgacgctc
     1201 ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa ttagtttgtt
     1261 ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc agcaagtata
     1321 tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat cctccaaaat
     1381 cggatctggt tccgcgtgga tctcgtcgtg catctgttgg atcctcaacg ccgattcctg
     1441 tcttgggtaa gccaacagac gctactggta atacaacatc taaatctgca ttttcattcg
     1501 gtactgctaa caccaatggt accaatgcct cagcaaactc cacatcattc tcgtttaacg
     1561 cccctgctac tggtaacggc acaactacta cttccaatac ctcaggaacc aatatagcgg
     1621 gtacatttaa cgtaggaaaa ccggatcaaa gtatcgctag tggcaatacg aatggagcgg
     1681 gctcggcatt tggcttttcg agctcaggaa cagcagcaac tggtgcagct tctaatcaat
     1741 cttcatttaa ttttggaaac aatggtgcag ggggtctcaa tccttttaca tcagcaactt
     1801 cgtcaactaa tgctaatgct ggtttattca ataaacctcc ttccacgaat gcacaaaatg
     1861 tcaatgttcc ctctgctttt aattttacgg gaaataattc aacgcctggt ggcggctctg
     1921 tatttaatat gaacggcaac actaatgcta atacggtgtt tgccggctct aataaccaac
     1981 cacatcaatc gcaaacccca tctttcaata caaacagctc attcacgcca tcaacagttc
     2041 ctaatattaa ttttagcgga ttgaatggcg gaattactaa taccgcgacc aaattcatcg
     2101 tgactgactg acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     2161 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     2221 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
     2281 cgatagcgga gtgtataatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     2341 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     2401 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2461 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     2521 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     2581 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     2641 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     2701 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     2761 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     2821 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     2881 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     2941 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3001 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     3061 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3121 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3181 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3241 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3301 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3361 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3421 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3481 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3541 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3601 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3661 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     3721 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3781 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3841 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3901 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3961 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4021 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4081 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4141 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     4201 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     4261 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     4321 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataaattccg acaccatcga
     4381 atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt
     4441 ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca
     4501 gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt
     4561 ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg
     4621 caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca
     4681 aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat
     4741 ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg
     4801 cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc
     4861 tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag
     4921 tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg
     4981 tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct
     5041 ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg
     5101 cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt
     5161 tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac
     5221 cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga
     5281 cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca
     5341 aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct
     5401 gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc
     5461 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     5521 cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca ccccaggctt
     5581 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa c
//
