LOCUS       sequence-153656-        7801 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express N-terminally FLAG tagged human chimera USP13/5-3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7801)
  AUTHORS   Liu Y, Soetandyo N, Lee JG, Liu L, Xu Y, Clemons WM Jr, Ye Y
  TITLE     USP13 antagonizes gp78 to maintain functionality of a chaperone in 
            ER-associated degradation.
  JOURNAL   Elife. 2014;3:e01369. doi: 10.7554/eLife.01369. Epub 2014 Jan 14.
  PUBMED    24424410
REFERENCE   2  (bases 1 to 7801)
  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: "2014"; volume: "3:e01369. doi"; pages: " 10.7554/eLife.01369.
            Epub 2014 Jan 14"
COMMENT     Sequence Label: PRK5-FLAG-USP13/5-3
FEATURES             Location/Qualifiers
     source          1..7801
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      918..927
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             927..950
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    1743..1754
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     promoter        complement(3592..3610)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3630..3647)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3636..3860
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3906..4334
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4343..4363)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4348..4678
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4528..4664
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4698..4715)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4698..4714)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4707..4729)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4927..5382
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5399..5418)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5518..5540
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5578..5596)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5664..5769
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5770..5838,5839..6630)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5988..6007)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6801..7389
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7290..7309
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7538..7555
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7672..7693
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7708..7713,7714..7731,7732..7738)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7746..7762
                     /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     7751..7773
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7770..7786
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7770..7786
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc taagcttgcc gccatggact acaaggacga cgatgacaag gggtcgacaa
      961 tggcggagct gagtgaggag gcgctgctgt cagtattacc gacgatccgg gtccctaagg
     1021 ctggagaccg ggtccacaaa gacgagtgcg ccttctcctt cgacacgccg gagtctgagg
     1081 ggggcctcta catctgtatg aacacgtttc tgggctttgg gaaacagtat gtggagagac
     1141 atttcaataa gaccggccag cgagtctact tgcacctccg gcggacccgg cgcccgaaag
     1201 aggaggaccc tgctacaggc actggagacc caccccggaa gaagcccacg cggctggcta
     1261 ttggtgttga aggcgggttt gaccttagcg aggagaagtt tgaattagac gaggatgtga
     1321 agattgtcat tttgccagat tacctggaga ttgcccggga tggactgggg ggactgcctg
     1381 acattgtcag agatcgggtg accagtgcag tggaggccct actgtcggcc gactcagcct
     1441 cccgcaagca ggaggtgcag gcatgggatg gggaagtacg gcaggtgtct aagcatgcct
     1501 tcagcctcaa gcagttggac aaccctgctc gaatccctcc ctgtggctgg aagtgctcca
     1561 agtgtgacat gagagagaac ctgtggctca acctgactga tggctccatc ctctgtgggc
     1621 gacgctactt cgatggcagt gggggcaaca accacgctgt ggagcactac cgagagacag
     1681 gctacccgtt agctgtcaag ctgggcacca tcacccctga tggagctgac gtgtactcat
     1741 atgatgagga tgacatggtc ctggacccca gcctggctga gcacctgtcc cacttcggca
     1801 tcgacatgct gaagatgcag aagacagaca agacgatgac tgagttggag atagacatga
     1861 accagcggat tggtgaatgg gagctgatcc aggagtcagg tgtgccactc aagcccctgt
     1921 ttgggcctgg ctacacaggc atccggaacc tgggtaacag ctgctacctc aactctgtgg
     1981 tccaggtgct cttcagcatc cctgacttcc agaggaagta tgtggataag ctggagaaga
     2041 tcttccagaa tgccccgacg gaccctaccc aggatttcag cacccaggtg gccaagctgg
     2101 gccatggcct tctctccggg gagtattcca agccagtacc ggagtcgggc gatggggagc
     2161 gggtgccaga acagaaggaa gttcaagatg gcattgcccc tcggatgttc aaggccctca
     2221 tcggcaaggg ccaccctgaa ttctccacca accggcagca ggatgcccag gagttcttcc
     2281 ttcaccttat caacatggtg gagaggaatt gccggagctc tgaaaatcct aatgaagtgt
     2341 tccgcttctt ggtggaggaa aagatcaagt gcctggccac agagaaggtg aagtacaccc
     2401 agcgagttga ctacatcatg cagctgcctg tgcccatgga tgcagccctt aacaaagagg
     2461 agcttctgga gtacgaggag aagaagcggc aagccgaaga ggagaagatg gcactgccag
     2521 aactggttcg ggcccaggtg cccttcagct cttgcctgga ggcctacggg gcccctgagc
     2581 aggtcgatga cttctggagc acggccctgc aggccaagtc agtagctgtc aagaccacac
     2641 gatttgcctc attccctgac tacctggtca tccagatcaa gaagttcacc ttcggcttag
     2701 actgggtgcc caagaaactg gatgtgtcca tcgagatgcc agaggagctc gacatctccc
     2761 agttgagggg cacagggctg cagcccggag aggaggagct gccagacatt gccccacccc
     2821 tggtcactcc ggatgagccc aaaggtagcc ttggtttcta tggcaacgaa gacgaagact
     2881 ccttctgctc ccctcacttc tcctctccga catcgcccat gatcgatgag tcatcagtga
     2941 tgcagctggc cgagatgggt ttcccgctgg aagcatgtcg caaggctgtg tacttcactg
     3001 gaaatatggg cgccgaggtg gccttcaact ggatcattgt tcacatggaa gagccagatt
     3061 ttgctgagcc gctgaccatg cctggttatg gaggggcagc ttctgctgga gcctctgttt
     3121 ttggtgcttc tggactggat aaccaacctc cagaggaaat cgtagctatc atcacctcca
     3181 tgggatttca gcgaaatcag gctattcagg cactacgagc aacgaataat aacctggaaa
     3241 gagcactgga ttggatcttt agccaccctg agtttgaaga agacagtgat tttgtgattg
     3301 agatggagaa taatgccaat gcaaacatta tttctgaggc caagcccgaa ggacctagag
     3361 tcaaggatgg atctggaaca tatgagctat ttgcattcat cagtcacatg ggaacatcca
     3421 caatgagtgg tcattacatt tgccatatca aaaaggaagg aagatgggtg atttacaatg
     3481 accacaaagt ttgtgcctca gaaaggcccc ctaaagacct gggctacatg tacttttacc
     3541 gcaggatacc aagctaagcg gccgctcgag catgcatcta gagggcccta ttctatagtg
     3601 tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     3661 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3721 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3781 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3841 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3901 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3961 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4021 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4081 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4141 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4201 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4261 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4321 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4381 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4441 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4501 accatagtca cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4561 tctccgcccc atggctgact aatttttttt tatttatgca gaggccgagg ccgcctcggc
     4621 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     4681 gctgttaaca gcttggcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     4741 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     4801 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
     4861 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     4921 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4981 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     5041 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5101 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     5161 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5221 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg
     5281 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     5341 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     5401 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     5461 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     5521 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     5581 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     5641 gtggcacttt tcggggaaat gtgcgcggaa cccctatttt gtttattttt ctaaatacat
     5701 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5761 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5821 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5881 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5941 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6001 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6061 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6121 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6181 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6241 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6301 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6361 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6421 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6481 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6541 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6601 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6661 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6721 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6781 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6841 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6901 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6961 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7021 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7081 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7141 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7201 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7261 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7321 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7381 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7441 gctcacatgt cttcctgcgt atccnngatt ctgtggatac cgtattaccg cctttgagtg
     7501 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     7561 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     7621 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     7681 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     7741 tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga ttacgaatta
     7801 a
//
