LOCUS       sequence-101060-        7768 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express N-terminally FLAG tagged human chimera USP13/5-6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7768)
  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 7768)
  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-USP13/5-6
FEATURES             Location/Qualifiers
     source          1..7768
                     /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"
     promoter        complement(3559..3577)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3597..3614)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3603..3827
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3873..4301
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4310..4330)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4315..4645
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4495..4631
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4665..4682)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4665..4681)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4674..4696)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4894..5349
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5366..5385)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5485..5507
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5545..5563)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5631..5736
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5737..5805,5806..6597)
                     /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(5955..5974)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6768..7356
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7257..7276
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7505..7522
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7639..7660
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7675..7680,7681..7698,7699..7705)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7713..7729
                     /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     7718..7740
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7737..7753
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7737..7753
                     /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 tgcagcgccg gggcgccctg ttcggcatgc cgggcggcag cggaggcagg aagatggctg
     1021 caggagacat cggcgagctg ctagtgcccc acatgcccac gatccgcgtg cccaggtccg
     1081 gcgacagggt ctacaagaac gagtgcgcct tctcctacga ctctcccaat tctgaaggtg
     1141 gactctatgt atgcatgaat acatttttgg cctttggaag ggaacatgtt gaaagacatt
     1201 ttcgaaaaac tggacagagt gtatacatgc acctgaaaag acatgtgcga gagaaggtaa
     1261 gaggggcgtc tggtggagcg ttaccaaaaa ggaggaattc caagattttt ttagatctag
     1321 atactgatga cgatttaaat agcgacgatt atgaatatga agatgaagcc aaacttgtta
     1381 tattcccaga tcactatgaa atagcactac caaatattga ggagttacca gccctggtaa
     1441 caattgcttg tgatgcagtt ctcagctcaa aatctccata cagaaagcag gacccagaca
     1501 cgtgggaaaa tgaattgcca gtatctaaat atgccaacaa cctcacccag ctggacaatg
     1561 gagtcaggat tcctccaagt ggttggaagt gtgccagatg cgacctgcga gaaaacctct
     1621 ggttgaatct gactgacggc tctgtcctgt gtggaaagtg gttctttgac agctctgggg
     1681 gcaacgggca tgcgctggag cattacagag acatgggcta cccactagcc gtgaaactgg
     1741 gaaccatcac tcctgacggg gcagatgttt attcttttca agaagaagaa cctgttttgg
     1801 atcctcattt ggccaagcac ttagcgcatt ttggaattga tatgcttcat atgcatggga
     1861 cagagaatgg gctccaggac aatgacatca agctgagggt cagtgagtgg gaagtgatcc
     1921 aggagtcggg cacgaaactg aagccaatgt atggtcctgg ctacacgggt ctgaagaacc
     1981 tgggcaacag ctgctatctc agctctgtca tgcaggccat cttcagcatc ccagaattcc
     2041 agagagcgta tgtaggaaac cttcccagaa tatttgacta ctcgccttta gatccaacac
     2101 aagatttcaa cacacagatg actaagttag gacatggcct tctctcaggc cagtattcaa
     2161 agcctccggt gaaatctgaa ctcattgaac aggtgatgaa ggaggagcac aagccacagc
     2221 agaacgggat ctctccgcgc atgtttaagg cctttgtaag caagagccac ccggaattct
     2281 cctctaacag gcagcaagat gcccaggaat tcttcttgca cctggtgaat ctagtagaga
     2341 ggaaccgcat cggctcagaa aacccaagcg atgtttttcg ttttttggtg gaagaacgca
     2401 ttcagtgctg tcagacccgg aaagtccgct acacggagag ggtggattac ctgatgcagt
     2461 tacctgtggc catggaggcg gcaaccaaca aggatgaact gatcgcttat gaactaacga
     2521 gaagggaagc agaagcaaac agaagacccc ttcctgagtt ggtacgtgcc aagataccat
     2581 ttagtgcctg ccttcaggcc ttctctgaac cagaaaatgt tgatgatttc tggagcagtg
     2641 ccctacaagc aaagtctgcg ggtgtgaaaa catctcgctt tgcttcattc cctgaatact
     2701 tggtagtgca gataaagaag ttcacttttg gtcttgactg ggttcccaaa aaatttgatg
     2761 tttctattga tatgccagac ctacttgata tcaaccatct ccgagccagg gggttacagc
     2821 caggagagga agaacttcca gacatcagcc cccccatagt cattcctgat gactcaaaag
     2881 atcgcctgat gaaccaattg atagacccat cagacatcga tgagtcatca gtgatgcagc
     2941 tggccgagat gggtttcccg ctggaagcat gtcgcaaggc tgtgtacttc actggaaata
     3001 tgggcgccga ggtggccttc aactggatca ttgttcacat ggaagagcca gattttgctg
     3061 agccgctgac catgcctggt tatggagggg cagcttctgc tggagcctct ccccctcctg
     3121 aggactgtgt gaccaccatt gtctccatgg gcttctcccg ggaccaggcc ttgaaagcgc
     3181 tgcgggccac gaacaatagt ttagaacggg ctgtggactg gatcttcagt cacattgacg
     3241 acctggatgc tgaagctgcc atggacatct cagagggccg ctcagctgcc gactccatct
     3301 ctgagtctgt gccagtggga cctaaagtcc gggatggtcc tggaaagtat cagctctttg
     3361 ccttcattag tcacatgggc acctctacca tgtgtggtca ctacgtctgc cacatcaaga
     3421 aagaaggcag atgggtgatc tacaatgacc agaaagtgtg tgcctccgag aagccgccca
     3481 aggacctggg ctacatctac ttctaccaga gagtggccag ctaagcggcc gctcgagcat
     3541 gcatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc
     3601 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3661 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3721 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3781 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     3841 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3901 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3961 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4021 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4081 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4141 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4201 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4261 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     4321 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4381 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4441 caaagcatgc atctcaatta gtcagcaacc atagtcacgc ccctaactcc gcccatcccg
     4501 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat ttttttttat
     4561 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     4621 ttttggaggc ctaggctttt gcaaaaagct gttaacagct tggcactggc cgtcgtttta
     4681 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     4741 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     4801 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     4861 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     4921 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4981 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     5041 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     5101 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     5161 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     5221 tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     5281 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     5341 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     5401 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     5461 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     5521 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     5581 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     5641 ctattttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5701 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5761 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5821 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5881 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5941 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6001 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6061 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6121 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6181 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6241 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6301 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6361 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6421 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6481 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6541 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6601 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6661 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6721 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6781 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6841 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6901 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6961 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7021 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7081 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7141 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7201 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7261 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7321 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7381 cggttcctgg ccttttgctg gccttttgct cacatgtctt cctgcgtatc cnngattctg
     7441 tggataccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     7501 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     7561 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     7621 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     7681 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     7741 aaacagctat gacatgatta cgaattaa
//
