LOCUS       sequence-368072-        6031 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of C-terminus of NUP1 nucleoporin as GST-Nup1-C
            fusion truncated so only one FXFG repeat remains.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6031)
  AUTHORS   Ken Belanger Lab
  TITLE     Belanger lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6031)
  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            "Unpublished"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: KBB96
FEATURES             Location/Qualifiers
     source          1..6031
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     946..965
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(985..1062)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(1135..1152)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1306..1894)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1386..1405)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2065..2856,2857..2925))
                     /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     2688..2707
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2926..3030)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3098..3116
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     3261..3283
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4429..4446)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(4453..5106)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(4473..4495)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     protein_bind    5129..5145
                     /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)."
     promoter        complement(join(5153..5159,5160..5175,5176..5181))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(5283..5810)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     5759..5781
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5773..5790
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5774..5790
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5806..5822)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5806..5822)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    5830..5846
                     /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)."
     promoter        complement(join(5854..5860,5861..5878,5879..5884))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     CDS             complement(join(5933..6031,1..984))
                     /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"
ORIGIN
        1 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
       61 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
      121 ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga
      181 gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
      241 ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt
      301 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
      361 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
      421 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
      481 tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
      541 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
      601 cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc
      661 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
      721 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
      781 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
      841 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
      901 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
      961 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
     1021 tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tcggaattta tgcggtgtga
     1081 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct
     1141 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     1201 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     1261 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1321 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1381 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1441 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1501 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1561 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1621 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1681 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1741 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1801 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1861 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1921 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1981 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2041 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2101 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     2161 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     2221 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     2281 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     2341 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg
     2401 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     2461 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     2521 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     2581 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     2641 atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc
     2701 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     2761 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     2821 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     2881 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     2941 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     3001 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     3061 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     3121 tcgtcttcaa gaattataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     3181 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     3241 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     3301 gaaacgcgcg aggcagatcg tcagtcagtc acgatgaatt tcaggtgtat tcttactggg
     3361 tatggaaaag gaaaaagatg ggtttgaatc attgctatca ctagactttt caggttttga
     3421 gttagaggtt gagttttcgt catttttttt ttcaattgtt gttgaaccaa acgtgaatga
     3481 gctcttttct agcgatggtt ttgcagctgc ttgaccaaat acaaaagatg gtttattatt
     3541 ttgagaatcc gcagcatgtt ttgttccaaa agtgaaggaa gcctcggctt tactgttgtt
     3601 gtccacattg gtcttggtag cttctgtcaa tgaaagggga tttttactct tcgtttgttc
     3661 agtagttgaa gtggccgaag aaaatgaaaa tggtttagca ttagtctcgg ttttcttttc
     3721 atttttatcg gtgcttggat tcagcttaaa tacatcactt gatacagttc cagaaaattt
     3781 aaattcaggg cccttgattt ctatatcgtt ttcgctatca ctttcatctt tcttgcgatt
     3841 ttcttttttt gggccaaatg agaaaacgga atctgaaata ggcggtttag catctattgt
     3901 gaacttgggc ccctccacct taacctcatc ttcagatgaa gaatcggaac ttttttggac
     3961 atcttctact ttggcaggtt tagcatcatt tccctttgtc tgcgttggat tctcgataaa
     4021 tttcacggga gcttctaaga tattctttat gtataattcg tacctggtga aaaggggcct
     4081 caagtccgct aacggcttgc ctagaactaa gtcatcaacc ttagccttga actggaggtt
     4141 caatgctttt agtctggaat tgctttctgt ggtagggcta ttatcaacct gagcttctcc
     4201 agtgccgtcc gcccggttca ggaagctaaa ggaactagcc tgcttggttt catccgattt
     4261 tgcagaacca aaaggtttga acgccatcct gcgctttggc atggcaattt ttcttctatt
     4321 catcacagca gatgacgcaa ccttagtgga gggagtcacg tcatcgtcag actcgttaga
     4381 atcgtacgtt tctctctgta tttgcgggga tccaacagat gcacgacgag atccacgcgg
     4441 aaccagatcc gattttggag gatggtcgcc accaccaaac gtggcttgcc agccctgcaa
     4501 aggccatgct atatacttgc tggatttcaa gtacttatca atttgtggga tagcttcaat
     4561 acgtttttta aaacaaacta attttgggaa cgcatccagg cacattgggt ccatgtataa
     4621 aacaacatca agagcgtcat acaacatgaa gtcaggatgg gttacatgat caccatttaa
     4681 atatgtttta tgacataaac gatcttcgaa cattttcagc atttcaggta gcttgctaag
     4741 aaaatcaact ttgagagttt caaagtcttt actatatgca attctcgaaa caccgtatct
     4801 aatatccaaa accgctcctt caagcattga aatctctgca cgctcttttg gacaaccacc
     4861 caacatgttg tgcttgtcag ctatataacg tatgatggcc atagactgtg ttaatttaac
     4921 atcaccatca atataataag gaagattggg aaactccaaa cccaattcaa actttttgtt
     4981 tcgccattta tcaccttcat cgcgctcata caaatgctct tcatattttt cttcaagata
     5041 ttccaaaaga agtcgagtgg gttgcacaag gcccttaatt ttccaataac ctagtatagg
     5101 ggacatgaat actgtttcct gtgtgaaatt gttatccgct cacaattcca cacattatac
     5161 gagccgatga ttaattgtca acagctcatt tcagaatatt tgccagaacc gttatgatgt
     5221 cggcgcaaaa aacattatcc agaacgggag tgcgccttga gcgacacgaa ttatgcagtg
     5281 atttacgacc tgcacagcca taccacagct tccgatggct gcctgacgcc agaagcattg
     5341 gtgcaccgtg cagtcgataa gctaattcca acgccatcaa aaataattcg cgtctggcct
     5401 tcctgtagcc agctttcatc aacattaaat gtgagcgagt aacaacccgt cggattctcc
     5461 gtgggaacaa acggcggatt gaccgtaatg ggataggtca cgttggtgta gatgggcgca
     5521 tcgtaaccgt gcatctgcca gtttgagggg acgacgacag tatcggcctc aggaagatcg
     5581 cactccagcc agctttccgg caccgcttct ggtgccggaa accaggcaaa gcgccattcg
     5641 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     5701 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     5761 cagtcacgac gttgtaaaac gacggccagt gaatccgtaa tcatggtcat agctgtttcc
     5821 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5881 taaagcctgg ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
     5941 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6001 gagaggcggt ttgcgtattg ggcgccaggg t
//
