LOCUS       sequence-399896-        5523 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GST fusion protein for quantification of gp78 levels.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5523)
  AUTHORS   Smith CE, Tsai YC, Liang YH, Khago D, Mariano J, Li J, Tarasov SG, 
            Gergel E, Tsai B, Villaneuva M, Clapp ME, Magidson V, Chari R, Byrd 
            RA, Ji X, Weissman AM
  TITLE     A structurally conserved site in AUP1 binds the E2 enzyme UBE2G2 and
            is essential for ER-associated degradation.
  JOURNAL   PLoS Biol. 2021 Dec 8;19(12):e3001474. doi: 
            10.1371/journal.pbio.3001474. eCollection 2021 Dec.
  PUBMED    34879065
REFERENCE   2  (bases 1 to 5523)
  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: "PLoS 
            Biol."; date: "2021-12-8"; volume: "19(12):e3001474. doi"; pages: " 
            10.1371/journal.pbio.3001474. eCollection 2021 Dec"
COMMENT     Sequence Label: pGEX-6P-1-gp78 429-611
FEATURES             Location/Qualifiers
     source          1..5523
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(191..196,197..212,213..219)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    227..243
                     /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)."
     CDS             266..919
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     877..899
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             926..949
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1581..1603)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1748..1766)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1834..1938
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1939..2007,2008..2799)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2157..2176)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2970..3558
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3459..3478
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3712..3729
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3802..3879
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3880..4962
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(3899..3918)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5011..5016,5017..5034,5035..5041)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5049..5065
                     /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     5073..5089
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5073..5089
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5085..5523,1..89)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5105..5122)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5105..5121)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5114..5136)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttggaattag cttatcgact gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg
       61 aagctgtggt atggctgtgc aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc
      121 actcccgttc tggataatgt tttttgcgcc gacatcataa cggttctggc aaatattctg
      181 aaatgagctg ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa
      241 caatttcaca caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg
      301 ccttgtgcaa cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt
      361 gtatgagcgc gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt
      421 tcccaatctt ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat
      481 acgttatata gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat
      541 ttcaatgctt gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag
      601 taaagacttt gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat
      661 gttcgaagat cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga
      721 cttcatgttg tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc
      781 gttcccaaaa ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta
      841 cttgaaatcc agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg
      901 tggcgaccat cctccaaaat cggatctgga agttctgttc caggggcccc tgggatccat
      961 tgcgagctgg ctgccgagtt tttcggttga agtgatgcac accaccaaca ttcttggcat
     1021 tacgcaggcc agcaactccc agctcaatgc aatggctcat cagattcaag agatgtttcc
     1081 ccaggtccca taccatctgg tactgcagga ccttcagctg acacgctcag ttgaaataac
     1141 aacagacaat attttagaag gacggattca agtacctttt cctacacagc ggtcagatag
     1201 catcagacct gcattgaaca gtcctgtgga aaggccaagc agtgaccagg aagagggaga
     1261 aacttctgct cagaccgagc gtgtgccact ggacctcagt cctcgcctgg aggagacgct
     1321 ggacttcggc gaggtggaag tggagcccag tgaggtggaa gacttcgagg ctcgtgggag
     1381 ccgcttctcc aagtctgctg atgagagaca gcgcatgctg gtgcagcgta aggacgaact
     1441 cctccagcaa gctcgcaaac gtttcttgaa caaaagttct gaagatgatg cggcctcaga
     1501 gagtttctaa gtcgactcga gcggccgcat cgtgactgac tgacgatctg cctcgcgcgt
     1561 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     1621 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     1681 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtataa ttcttgaaga
     1741 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     1801 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     1861 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1921 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1981 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     2041 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     2101 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     2161 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     2221 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     2281 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     2341 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     2401 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     2461 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     2521 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2581 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2641 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2701 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2761 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2821 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2881 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2941 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3001 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3061 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     3121 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3181 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3241 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3301 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3361 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3421 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3481 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3541 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3601 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3661 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3721 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3781 atttcacacc gcataaattc cgacaccatc gaatggcgca aaacctttcg cggtatggca
     3841 tgatagcgcc cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac
     3901 gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc
     3961 agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac
     4021 attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc
     4081 acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc
     4141 gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt
     4201 aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg
     4261 ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt
     4321 cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg
     4381 cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc
     4441 ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc
     4501 aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa
     4561 caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat
     4621 cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat
     4681 atctcggtag tgggatacga cgataccgaa gacagctcat gttatatccc gccgttaacc
     4741 accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc
     4801 tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa
     4861 accacgctag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     4921 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     4981 aagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     5041 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgg
     5101 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     5161 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     5221 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgctttgcc tggtttccgg
     5281 caccagaagc ggtgccggaa agctggctgg agtgcgatct tcctgaggcc gatactgtcg
     5341 tcgtcccctc aaactggcag atgcacggtt acgatgcgcc catctacacc aacgtgacct
     5401 atcccattac ggtcaatccg ccgtttgttc ccacggagaa tccgacgggt tgttactcgc
     5461 tcacatttaa tgttgatgaa agctggctac aggaaggcca gacgcgaatt atttttgatg
     5521 gcg
//
