LOCUS       sequence-430246-        8258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of Importin-5 tagged on its N-terminus with 
            GST. Cleavable with 3C protease..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8258)
  AUTHORS   Pardal AJ, Bowman AJ
  TITLE     A specific role for importin-5 and NASP in the import and nuclear 
            hand-off of monomeric H3.
  JOURNAL   Elife. 2022 Sep 6;11:e81755. doi: 10.7554/eLife.81755.
  PUBMED    36066346
REFERENCE   2  (bases 1 to 8258)
  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: "Elife."; 
            date: "2022-09-6"; volume: "11:e81755. doi"; pages: " 
            10.7554/eLife.81755"
COMMENT     Sequence Label: pCL1324
FEATURES             Location/Qualifiers
     source          1..8258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(708..730)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(875..893)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        961..1065
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1066..1134,1135..1926)
                     /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(1284..1303)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2097..2685
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2586..2605
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2839..2856
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        2929..3006
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3007..4089
                     /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(3026..3045)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4138..4143,4144..4161,4162..4168)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4176..4192
                     /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     4200..4216
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4200..4216
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4212..4739
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(4232..4249)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4232..4248)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4241..4263)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(4841..4846,4847..4862,4863..4869)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4877..4893
                     /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             4916..5569
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     5527..5549
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             5576..5599
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
ORIGIN
        1 ggggattatg catctttgat gatgtcatag aacactgtag tccagcctca tttaaatacg
       61 cagaatattt cttaagacca atgctccaat atgtatgtga caacagccca gaagtcaggc
      121 aagcagctgc atatggcctg ggagtcatgg cacagtacgg tggagataat tatcgccctt
      181 tttgtacaga agcacttccc ctgctggtaa gagttattca gtctgcggat tctaagacca
      241 aagaaaatgt caatgctaca gagaactgca tctcagcagt agggaaaatc atgaagttca
      301 agcctgactg tgtaaacgtt gaagaggtcc ttccacactg gttgtcttgg cttccactac
      361 atgaagataa agaagaagct gttcagactt tcaattatct gtgtgacctg attgaaagta
      421 atcatccaat tgttcttggc ccaaacaata ccaatctgcc caaaatattt agtataattg
      481 cggaaggaga aatgcacgag gcaattaaac atgaagatcc ttgtgccaaa cgtctggcca
      541 atgtcgttcg ccaagtacag acttctggag gactgtggac tgagtgcata gcacagctca
      601 gtcctgagca gcaggccgcc attcaggagc tcctgaactc tgcgtgagcg gccgcatcgt
      661 gactgactga cgatctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
      721 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
      781 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
      841 gatagcggag tgtataattc ttgaagacga aagggcctcg tgatacgcct atttttatag
      901 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
      961 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     1021 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     1081 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     1141 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     1201 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     1261 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     1321 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     1381 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     1441 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     1501 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     1561 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     1621 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     1681 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     1741 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     1801 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     1861 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     1921 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     1981 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     2041 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2101 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2161 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2221 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     2281 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2341 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     2401 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2461 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2521 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2581 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2641 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2701 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2761 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2821 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     2881 tattttctcc ttacgcatct gtgcggtatt tcacaccgca taaattccga caccatcgaa
     2941 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
     3001 gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag
     3061 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg
     3121 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc
     3181 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa
     3241 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg
     3301 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc
     3361 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct
     3421 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt
     3481 attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt
     3541 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg
     3601 gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc
     3661 gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt
     3721 cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc
     3781 gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac
     3841 agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa
     3901 accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg
     3961 ttgcccgtct cactggtgaa aagaaaaacc acgctagcgc ccaatacgca aaccgcctct
     4021 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     4081 gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cccaggcttt
     4141 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     4201 aggaaacagc tatgaccatg attacggatt cactggccgt cgttttacaa cgtcgtgact
     4261 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     4321 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     4381 gcgaatggcg ctttgcctgg tttccggcac cagaagcggt gccggaaagc tggctggagt
     4441 gcgatcttcc tgaggccgat actgtcgtcg tcccctcaaa ctggcagatg cacggttacg
     4501 atgcgcccat ctacaccaac gtgacctatc ccattacggt caatccgccg tttgttccca
     4561 cggagaatcc gacgggttgt tactcgctca catttaatgt tgatgaaagc tggctacagg
     4621 aaggccagac gcgaattatt tttgatggcg ttggaattag cttatcgact gcacggtgca
     4681 ccaatgcttc tggcgtcagg cagccatcgg aagctgtggt atggctgtgc aggtcgtaaa
     4741 tcactgcata attcgtgtcg ctcaaggcgc actcccgttc tggataatgt tttttgcgcc
     4801 gacatcataa cggttctggc aaatattctg aaatgagctg ttgacaatta atcatcggct
     4861 cgtataatgt gtggaattgt gagcggataa caatttcaca caggaaacag tattcatgtc
     4921 ccctatacta ggttattgga aaattaaggg ccttgtgcaa cccactcgac ttcttttgga
     4981 atatcttgaa gaaaaatatg aagagcattt gtatgagcgc gatgaaggtg ataaatggcg
     5041 aaacaaaaag tttgaattgg gtttggagtt tcccaatctt ccttattata ttgatggtga
     5101 tgttaaatta acacagtcta tggccatcat acgttatata gctgacaagc acaacatgtt
     5161 gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt gaaggagcgg ttttggatat
     5221 tagatacggt gtttcgagaa ttgcatatag taaagacttt gaaactctca aagttgattt
     5281 tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat cgtttatgtc ataaaacata
     5341 tttaaatggt gatcatgtaa cccatcctga cttcatgttg tatgacgctc ttgatgttgt
     5401 tttatacatg gacccaatgt gcctggatgc gttcccaaaa ttagtttgtt ttaaaaaacg
     5461 tattgaagct atcccacaaa ttgataagta cttgaaatcc agcaagtata tagcatggcc
     5521 tttgcagggc tggcaagcca cgtttggtgg tggcgaccat cctccaaaat cggatctgga
     5581 agttctgttc caggggcccc tgggatccgc aatggcggcg gctgcggcgg agcagcaaca
     5641 gttctacctg ctcctgggaa acctgctcag ccccgacaat gtggtccgga aacaggcaga
     5701 ggaaacctat gagaatatcc caggccagtc aaagatcaca ttcctcttac aagccatcag
     5761 aaatacaaca gctgctgaag aggctagaca aatggccgcc gttctcctaa gacgtctctt
     5821 gtcctctgca tttgatgaag tctatccagc acttccctct gatgttcaga ctgccatcaa
     5881 gagtgagcta ctcatgatta ttcagatgga aacacaatct agcatgagga aaaaagtttg
     5941 tgatattgcg gcagaactgg ccaggaattt aatagatgag gatggcaata accagtggcc
     6001 cgaaggtttg aagttccttt ttgattcagt cagctctcaa aatgtgggac tgcgggaagc
     6061 tgcccttcac attttctgga actttcctgg aatttttggg aaccagcaac aacactattt
     6121 agatgtcatc aaacgaatgt tagttcagtg tatgcaagat caggaacacc cgtcgatcag
     6181 gacgttatct gctagagcta cagctgcatt tatacttgca aatgagcata atgttgctct
     6241 gttcaaacat tttgcagact tgctaccggg attcctacag gcggtaaatg actcgtgcta
     6301 ccagaatgat gattctgtcc taaaatccct cgttgagatt gcagatactg ttccaaagta
     6361 tttgcgtcct cacttggaag caactctaca gctaagtcta aagttgtgtg gagacactag
     6421 cctcaacaat atgcaacgcc agcttgccct tgaagtgatc gtcaccctct ctgagactgc
     6481 agctgctatg ttaagaaaac ataccaatat tgttgcacag actattcctc agatgttagc
     6541 aatgatggtt gatttggaag aagatgagga ctgggcaaat gcagatgaac tagaagatga
     6601 tgattttgac agcaatgcag ttgcaggcga gagtgctcta gatcgaatgg cttgtggact
     6661 tggtggaaag ctcgttctgc cgatgatcaa ggaacacatt atgcaaatgc ttcaaaatcc
     6721 tgactggaaa taccggcatg caggattgat ggccttatct gccattggtg aagggtgcca
     6781 ccagcaaatg gaaggaattc taaatgagat cgtaaatttt gttttacttt ttctccagga
     6841 tcctcatcca agagtaaggt atgcagcctg taatgccgtg ggacagatgg ctacagattt
     6901 tgcacctggt ttccaaaaga aatttcatga gaaggtgatt gcagctctgc tgcagaccat
     6961 ggaagaccaa ggcaatcaac gtgtgcaggc ccatgcagct gctgccctca ttaactttac
     7021 tgaagactgt cccaagtcac tacttattcc atacttggat aatttggtga aacatctgca
     7081 ttccattatg gtactgaagc ttcaagagct gattcagaaa ggcaccaagt tagttttgga
     7141 acaagttgtg acatccattg catcagttgc cgatactgca gaagaaaaat ttgtccccta
     7201 ctatgattta tttatgccat cactgaagca catcgttgag aatgcggttc aaaaagaact
     7261 gagacttctg agaggaaaaa ctattgaatg cattagcctc attggtctgg ctgttgggaa
     7321 ggaaaaattc atgcaggatg catcagatgt gatgcagctt ttgttaaaga cccagacaga
     7381 cttcaatgat atggaagatg atgatcctca gatctcttac atgatctcag catgggccag
     7441 aatgtgcaaa atccttggaa aagaatttca gcaatacctt ccagtggtta tggggccttt
     7501 aatgaagacg gcttcaatta agcccgaagt agccctttta gatacccaag acatggagaa
     7561 tatgagtgat gatgatggtt gggaatttgt gaaccttgga gatcagcaaa gctttggtat
     7621 taaaactgca ggactagaag aaaaatcaac tgcttgccag atgttggttt gctatgctaa
     7681 ggagttaaag gaaggctttg tggagtacac cgaacaggtt gtcaaactga tggtcccttt
     7741 actgaaattt tatttccacg atggtgttcg agtggcagca gcggaatcca tgcctcttct
     7801 cctggagtgt gcaagagtcc gtggtcctga gtatctcaca cagatgtggc attttatgtg
     7861 tgatgctcta attaaggcca ttggtacaga accagattca gacgtcctct cagaaataat
     7921 gcattctttt gcaaagtgca ttgaagtaat gggagatgga tgccttaata atgaacactt
     7981 tgaagaactg ggaggtatat tgaaagcaaa gcttgaagaa cattttaaaa atcaagaatt
     8041 acgacaagtt aaaagacaag atgaagacta tgatgaacag gtcgaagagt cactacaaga
     8101 tgaggatgat aatgatgttt atattctgac caaagtgtca gatattttac actcaatatt
     8161 cagtagctac aaagaaaagg tgttaccatg gtttgaacag ctgcttccat taattgtcaa
     8221 cctcatttgt ccacatagac catggccaga cagacaat
//
