LOCUS       sequence-101024-        7759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express N-terminally FLAG tagged human chimera USP13/5-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7759)
  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 7759)
  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-USP13/5-1
FEATURES             Location/Qualifiers
     source          1..7759
                     /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(3550..3568)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3588..3605)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3594..3818
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3864..4292
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4301..4321)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4306..4636
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4486..4622
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4656..4673)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4656..4672)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4665..4687)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4885..5340
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5357..5376)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5476..5498
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5536..5554)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5622..5727
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5728..5796,5797..6588)
                     /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(5946..5965)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6759..7347
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7248..7267
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7496..7513
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7630..7651
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7666..7671,7672..7689,7690..7696)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7704..7720
                     /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     7709..7731
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7728..7744
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7728..7744
                     /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 gcagctggac aatggagtca ggattcctcc aagtggttgg aagtgtgcca
     1561 gatgcgacct gcgagaaaac ctctggttga atctgactga cggctctgtc ctgtgtggaa
     1621 agtggttctt tgacagctct gggggcaacg ggcatgcgct ggagcattac agagacatgg
     1681 gctacccact agccgtgaaa ctgggaacca tcactcctga cggggcagat gtttattctt
     1741 ttcaagaaga agaacctgtt ttggatcctc atttggccaa gcacttagcg cattttggaa
     1801 ttgatatgct tcatatgcat gggacagaga atgggctcca ggacaatgac atcaagctga
     1861 gggtcagtga gtgggaagtg atccaggagt cgggcacgaa actgaagcca atgtatggtc
     1921 ctggctacac gggtctgaag aacctgggca acagctgcta tctcagctct gtcatgcagg
     1981 ccatcttcag catcccagaa ttccagagag cgtatgtagg aaaccttccc agaatatttg
     2041 actactcgcc tttagatcca acacaagatt tcaacacaca gatgactaag ttaggacatg
     2101 gccttctctc aggccagtat tcaaagcctc cggtgaaatc tgaactcatt gaacaggtga
     2161 tgaaggagga gcacaagcca cagcagaacg ggatctctcc gcgcatgttt aaggcctttg
     2221 taagcaagag ccacccggaa ttctcctcta acaggcagca agatgcccag gaattcttct
     2281 tgcacctggt gaatctagta gagaggaacc gcatcggctc agaaaaccca agcgatgttt
     2341 ttcgtttttt ggtggaagaa cgcattcagt gctgtcagac ccggaaagtc cgctacacgg
     2401 agagggtgga ttacctgatg cagttacctg tggccatgga ggcggcaacc aacaaggatg
     2461 aactgatcgc ttatgaacta acgagaaggg aagcagaagc aaacagaaga ccccttcctg
     2521 agttggtacg tgccaagata ccatttagtg cctgccttca ggccttctct gaaccagaaa
     2581 atgttgatga tttctggagc agtgccctac aagcaaagtc tgcgggtgtg aaaacatctc
     2641 gctttgcttc attccctgaa tacttggtag tgcagataaa gaagttcact tttggtcttg
     2701 actgggttcc caaaaaattt gatgtttcta ttgatatgcc agacctactt gatatcaacc
     2761 atctccgagc cagggggtta cagccaggag aggaagaact tccagacatc agccccccca
     2821 tagtcattcc tgatgactca aaagatcgcc tgatgaacca attgatagac ccatcagaca
     2881 tcgatgagtc atcagtgatg cagctggccg agatgggttt cccgctggaa gcatgtcgca
     2941 aggctgtgta cttcactgga aatatgggcg ccgaggtggc cttcaactgg atcattgttc
     3001 acatggaaga gccagatttt gctgagccgc tgaccatgcc tggttatgga ggggcagctt
     3061 ctgctggagc ctctgttttt ggtgcttctg gactggataa ccaacctcca gaggaaatcg
     3121 tagctatcat cacctccatg ggatttcagc gaaatcaggc tattcaggca ctacgagcaa
     3181 cgaataataa cctggaaaga gcactggatt ggatctttag ccaccctgag tttgaagaag
     3241 acagtgattt tgtgattgag atggagaata atgccaatgc aaacattatt tctgaggcca
     3301 agcccgaagg acctagagtc aaggatggat ctggaacata tgagctattt gcattcatca
     3361 gtcacatggg aacatccaca atgagtggtc attacatttg ccatatcaaa aaggaaggaa
     3421 gatgggtgat ttacaatgac cacaaagttt gtgcctcaga aaggccccct aaagacctgg
     3481 gctacatgta cttttaccgc aggataccaa gctaagcggc cgctcgagca tgcatctaga
     3541 gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc
     3601 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3661 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3721 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3781 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3841 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3901 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3961 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4021 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4081 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4141 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4201 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4261 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4321 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4381 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4441 catctcaatt agtcagcaac catagtcacg cccctaactc cgcccatccc gcccctaact
     4501 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttta tttatgcaga
     4561 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4621 cctaggcttt tgcaaaaagc tgttaacagc ttggcactgg ccgtcgtttt acaacgtcgt
     4681 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     4741 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     4801 aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4861 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4921 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4981 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5041 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5101 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5161 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     5221 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     5281 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     5341 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     5401 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     5461 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     5521 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     5581 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctattttgt
     5641 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5701 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5761 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5821 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5881 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5941 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6001 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6061 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6121 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6181 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6241 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6301 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6361 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6421 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6481 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6541 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6601 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6661 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6721 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6781 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6841 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6901 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6961 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7021 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7081 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7141 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7201 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7261 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7321 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7381 gccttttgct ggccttttgc tcacatgtct tcctgcgtat ccnngattct gtggataccg
     7441 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     7501 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     7561 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     7621 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     7681 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     7741 tgacatgatt acgaattaa
//
