LOCUS       sequence-352597-        5767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human UBQLN2 construct 450-624 with added C-terminal W in 
            bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5767)
  AUTHORS   Dao TP, Kolaitis RM, Kim HJ, O'Donovan K, Martyniak B, Colicino E, 
            Hehnly H, Taylor JP, Castaneda CA
  TITLE     Ubiquitin Modulates Liquid-Liquid Phase Separation of UBQLN2 via 
            Disruption of Multivalent Interactions.
  JOURNAL   Mol Cell. 2018 Mar 15;69(6):965-978.e6. doi: 
            10.1016/j.molcel.2018.02.004. Epub 2018 Mar 8.
  PUBMED    29526694
REFERENCE   2  (bases 1 to 5767)
  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: 
            "10.1016/j.molcel.2018.02.004"; journalName: "Mol Cell"; date: 
            "2018-03-15- 15"; volume: "69"; issue: "6"; pages: "965-978.e6"
COMMENT     Sequence Label: UBQLN2_450-624W
FEATURES             Location/Qualifiers
     source          1..5767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    86..225
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     141..160
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(240..257)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(411..999)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(491..510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1110..1129)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1121..1936
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             1121..1936
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1194..1213)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(2028..2483)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2165..2186)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2377..2396
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(2520..2567)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2563..2581
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(2634..2651)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             3199..3221
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    3236..3260
                     /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     complement(3260..3279)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3261..3279)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3363..3382)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        3588..3665
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3666..4748
                     /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(3685..3704)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             5560..5751
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(5745..5767)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
       61 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
      121 tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accattgcgg
      181 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc
      241 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
      301 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
      361 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
      421 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
      481 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      541 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      601 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      661 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      721 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      781 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      841 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
      901 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
      961 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1021 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgaacaat
     1081 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     1141 aacgtcttgc tctaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     1201 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc
     1261 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     1321 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     1381 tatccgtact cctgatgatg catggttact caccactgcg atccccggga aaacagcatt
     1441 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     1501 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     1561 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     1621 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     1681 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     1741 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     1801 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     1861 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     1921 cgatgagttt ttctaagaat taattcatga gcggatacat atttgaatgt atttagaaaa
     1981 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt
     2041 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     2101 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     2161 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     2221 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     2281 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     2341 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     2401 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     2461 taatgcgccg ctacagggcg cgtcccattc gccaatccgg atatagttcc tcctttcagc
     2521 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctagtta ttgctcagcg
     2581 gtggcagcag ccaactcagc ttcctttcgg gctttgttag cagccggatc tcagtggtgg
     2641 tggtggtggt gctcgagtta ccacgatggc tgggagccca gcagcctttc aatggctgca
     2701 ttgatgtcgc ctcctgttgc tattagggcc tgcaagtttg cttcacggtt taagaacccc
     2761 attgcgttga gctgttccag ttgttgctga aatctgactt ctggattcgg cagctgtgga
     2821 gcatttgctc cagccagggc ctgcaccatt tgctgaatga actgctggtt gggtccagat
     2881 tctgatgtag gactcgtggt ttcactaggt gcagcgctgg acacagtagg ccccgtgggg
     2941 ccaccagagc cggtggagcc agggggggct gcagggccag tgggtcctat gggcccaatg
     3001 gggcctatgg gggtaaaagg gactataggg cctatggggc ctatgggggt gactgggcct
     3061 acagggccta tagcggttcc cagcaccccc acccccacac ctggagtgaa gctcggaatc
     3121 aggccaggtg cttcagtggc taatgtctgt agcccctgct ggatctgcat taaagcctgc
     3181 attgctctca tatgtatatc tccttcttaa agttaaacaa aattatttct agaggggaat
     3241 tgttatccgc tcacaattcc cctatagtga gtcgtattaa tttcgcggga tcgagatctc
     3301 gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg
     3361 cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag
     3421 cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat
     3481 ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg
     3541 ctgcttccta atgcaggagt cgcataaggg agagcgtcga gatcccggac accatcgaat
     3601 ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg
     3661 tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga
     3721 ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg
     3781 aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca
     3841 aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa
     3901 ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg
     3961 tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg
     4021 tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg
     4081 cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta
     4141 ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc
     4201 accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg
     4261 ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggag cgggaaggcg
     4321 actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc
     4381 ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg
     4441 agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca
     4501 gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa
     4561 ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt
     4621 tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc
     4681 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     4741 ggcagtgagc gcaacgcaat taatgtaagt tagctcactc attaggcacc gggatctcga
     4801 ccgatgccct tgagagcctt caacccagtc agctccttcc ggtggagccc gcggggcatg
     4861 actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg
     4921 gcagcgctct gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc
     4981 ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc
     5041 gccaccaaac gtttcggcga gaagcaggcc attatcgccg gcatggcggc cccacgggtg
     5101 cgcatgatcg tgctcctgtc gttgaggacc cggctaggct ggcggggttg ccttactggt
     5161 tagcagaatg aatcaccgat acgcgagcga acgtgaagcg actgctgctg caaaacgtct
     5221 gcgacctgag caacaacatg aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg
     5281 cggaagtcag cgccctgcac cattatgttc cggatctgca tcgcaggatg ctgctggcta
     5341 ccctgtggaa cacctacatc tgtattaacg aagcgctggc attgaccctg agtgattttt
     5401 ctctggtccc gccgcatcca taccgccagt tgtttaccct cacaacgttc cagtaaccgg
     5461 gcatgttcat catcagtaac ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt
     5521 acccccatga acagaaatcc cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc
     5581 gcccttaaca tggcccgctt tatcagaagc cagacattaa cgcttctgga gaaactcaac
     5641 gagctggacg cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag
     5701 ctttaccgca gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     5761 ctcccgg
//
