LOCUS       sequence-424666-        7379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express C-terminally FLAG tagged gp78.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7379)
  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 7379)
  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: pcDNA-gp78-FLAG
FEATURES             Location/Qualifiers
     source          1..7379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     26..45
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             180..974
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(234..253)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     844..863
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1148..1269
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1185..1204)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1239..1258
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1318..1334)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1318..1334)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1331..1353)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1342..1358
                     /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)."
     promoter        complement(join(1366..1372,1373..1390,1391..1396))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1411..1432
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1549..1566)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1720..2308)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1800..1819)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2479..3270,3271..3339))
                     /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     3102..3121
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3340..3444)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3519..3538)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3710..4089
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4090..4293
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4244..4264
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4338..4357
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4338..4356
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6393..6416
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(6445..6462)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6451..6675
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6721..7149
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6808..6827)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7018..7039
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7158..7178)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(7163..7379,1..113)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(7343..7379,1..99)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
       61 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
      121 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
      181 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
      241 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
      301 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
      361 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
      421 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
      481 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
      541 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
      601 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
      661 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
      721 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
      781 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
      841 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
      901 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
      961 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     1021 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     1081 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     1141 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     1201 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     1261 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     1321 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     1381 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     1441 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     1501 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     1561 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1621 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1681 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1741 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1801 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1861 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1921 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1981 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2041 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2101 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2161 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2221 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2281 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2341 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2401 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2461 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2521 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     2581 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     2641 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     2701 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     2761 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     2821 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     2881 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     2941 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3001 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3061 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3121 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3181 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3241 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3301 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3361 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3421 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     3481 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
     3541 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     3601 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
     3661 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
     3721 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     3781 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     3841 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     3901 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     3961 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     4021 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     4081 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     4141 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     4201 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     4261 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
     4321 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
     4381 aacgggccct ctagactcga gcggccgcca ctgtgctgga tatctgcaga attcggcttg
     4441 ccgccatgcc gctgctcttc ctcgagcgct tcccctggcc cagcctccgc acctacacgg
     4501 gcctcagcgg cctggccctg ctgggcacca tcatcagcgc ctaccgcgcg ctcagccagc
     4561 ccgaggccgg ccccggcgag ccggaccagc taacggcctc gctgcagcct gagccgccgg
     4621 cgcccgcccg gccgagcgcc gggggacccc gggcccgcga tgtggcccag tacctgctct
     4681 cagacagcct cttcgtgtgg gttctagtaa ataccgcttg ctgtgttttg atgttggtgg
     4741 ctaagctcat ccagtgtatt gtgtttggcc ctcttcgagt gagtgagaga cagcatctca
     4801 aagacaaatt ttggaatttt attttctaca agttcatttt catctttggt gtgctgaatg
     4861 tccagacagt ggaggaggtg gtcatgtggt gcctctggtt tgccggactt gtctttctgc
     4921 acctgatggt tcagctctgc aaggatcgat ttgaatatct ttccttctcg cccaccacgc
     4981 tgatgagcag ccacggtcga gtcctgtccc tgttggttgc catgctgctt tcctgctgtg
     5041 gactggcggc cgtctgctcc atcaccggct acacccacgg aatgcacacc ttggctttca
     5101 tggctgcaga gtctcttctt gtgacagtga ggactgctca tgtgatttta cgatacgtaa
     5161 ttcacctctg ggacctcaac cacgaaggga cgtgggaagg aaaggggacg tatgtctatt
     5221 acacagactt agtcatggag ctcactctcc tgtccctgga cctcatgcac catattcaca
     5281 tgttgttatt tggcaacatc tggttatcca tggccagcct ggtcatcttt atgcagctgc
     5341 gttacctgtt tcatgaggtg caacgtcgaa ttcgtcggca caagaactgt ctacgtgtgg
     5401 ttggaaacat ggaggccagg tttgcagttg caactccaga ggagctggct gtcaacaatg
     5461 acgactgtgc catctgttgg ggctccatgc aggctgcgcg gaaactgccc tgtggacatc
     5521 ttttccacaa ctcctgtctt cgttcctggc tagaacaaga cacctcctgt ccaacatgca
     5581 gaatgtctct taatattgcc gacaataatc gtgtcaggga agaacatcaa ggagagaact
     5641 tggatgagaa tttggttcct gtagcagcag ccgaagggag acctcgctta aaccaacaca
     5701 atcacttctt ccatttcgat gggtctcgga ttgcgagctg gctgccgagt ttttcggttg
     5761 aagtgatgca caccaccaac attcttggca ttacgcaggc cagcaactcc cagctcaatg
     5821 caatggctca tcagattcaa gagatgtttc cccaggtccc ataccatctg gtactgcagg
     5881 accttcagct gacacgctca gttgaaataa caacagacaa tattttagaa ggacggattc
     5941 aagtaccttt tcctacacag cggtcagata gcatcagacc tgcattgaac agtcctgtgg
     6001 aaaggccaag cagtgaccag gaagagggag aaacttctgc tcagaccgag cgtgtgccac
     6061 tggacctcag tcctcgcctg gaggagacgc tggacttcgg cgaggtggaa gtggagccca
     6121 gtgaggtgga agacttcgag gctcgtggga gccgcttctc caagtctgct gatgagagac
     6181 agcgcatgct ggtgcagcgt aaggacgaac tcctccagca agctcgcaaa cgtttcttga
     6241 acaaaagttc tgaagatgat gcggcctcag agagcttcct cccctcggaa ggtgcgtcct
     6301 ctgaccccgt gaccctgcgt cgaaggatgc tggctgccgc cgcggaacgg aggcttcaga
     6361 agcagcagac ctccgaattc ggatccggta ccgattacaa ggatgacgac gataagtgaa
     6421 gcttaagttt aaaccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     6481 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     6541 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     6601 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     6661 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     6721 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     6781 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     6841 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     6901 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     6961 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     7021 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     7081 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     7141 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     7201 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     7261 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     7321 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctc
//
