LOCUS       sequence-100804-        7795 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express N-terminally FLAG tagged human USP13.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7795)
  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 7795)
  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
FEATURES             Location/Qualifiers
     source          1..7795
                     /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(3586..3604)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3624..3641)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3630..3854
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3900..4328
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4337..4357)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4342..4672
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4522..4658
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4692..4709)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4692..4708)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4701..4723)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4921..5376
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5393..5412)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5512..5534
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5572..5590)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5658..5763
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5764..5832,5833..6624)
                     /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(5982..6001)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6795..7383
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7284..7303
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7532..7549
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7666..7687
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7702..7707,7708..7725,7726..7732)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7740..7756
                     /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     7745..7767
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7764..7780
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7764..7780
                     /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 gtttttggtg
     3121 cttctggact ggataaccaa cctccagagg aaatcgtagc tatcatcacc tccatgggat
     3181 ttcagcgaaa tcaggctatt caggcactac gagcaacgaa taataacctg gaaagagcac
     3241 tggattggat ctttagccac cctgagtttg aagaagacag tgattttgtg attgagatgg
     3301 agaataatgc caatgcaaac attatttctg aggccaagcc cgaaggacct agagtcaagg
     3361 atggatctgg aacatatgag ctatttgcat tcatcagtca catgggaaca tccacaatga
     3421 gtggtcatta catttgccat atcaaaaagg aaggaagatg ggtgatttac aatgaccaca
     3481 aagtttgtgc ctcagaaagg ccccctaaag acctgggcta catgtacttt taccgcagga
     3541 taccaagcta agcggccgct cgagcatgca tctagagggc cctattctat agtgtcacct
     3601 aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3661 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3721 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3781 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3841 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3901 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3961 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4021 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4081 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4141 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4201 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4261 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4321 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4381 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4441 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4501 gtcacgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4561 ccccatggct gactaatttt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4621 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctgtt
     4681 aacagcttgg cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     4741 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     4801 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg
     4861 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     4921 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4981 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     5041 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     5101 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     5161 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     5221 gactcttgtt ccaaactgga acaacactca accctatctc gggctattct tttgatttat
     5281 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     5341 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     5401 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     5461 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     5521 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     5581 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     5641 cttttcgggg aaatgtgcgc ggaaccccta ttttgtttat ttttctaaat acattcaaat
     5701 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5761 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5821 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5881 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5941 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     6001 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     6061 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     6121 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6181 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     6241 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6301 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6361 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6421 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6481 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6541 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6601 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6661 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6721 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6781 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6841 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6901 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6961 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7021 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     7081 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7141 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7201 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7261 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7321 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7381 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7441 atgtcttcct gcgtatccnn gattctgtgg ataccgtatt accgcctttg agtgagctga
     7501 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     7561 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     7621 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     7681 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     7741 tgtgagcgga taacaatttc acacaggaaa cagctatgac atgattacga attaa
//
