LOCUS       sequence-410484-        7498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Assessing  cellular requirement for G2BRgp78.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7498)
  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 7498)
  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: pCINeo-gp78 Mutant G2BR
FEATURES             Location/Qualifiers
     source          1..7498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3202..3323
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3213..3232)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3267..3286
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3509..3964
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3596..3615)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3806..3827
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4081..4438
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4081..4272
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4104..4124)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4289..4424
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4489..5283
                     /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(4543..4562)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5153..5172
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5347..5395
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5436..5455)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5555..5577
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5615..5633)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5701..5805
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5806..5874,5875..6666)
                     /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(6024..6043)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6837..7425
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7326..7345
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc ctcgagaatt cacgcgtggt acctctagat gcatgctcga gcggccgcca
     1141 gtgtgatgga tatctgcaga attcggcttg ccgccatgcc gctgctcttc ctcgagcgct
     1201 tcccctggcc cagcctccgc acctacacgg gcctcagcgg cctggccctg ctgggcacca
     1261 tcatcagcgc ctaccgcgcg ctcagccagc ccgaggccgg ccccggcgag ccggaccagc
     1321 taacggcctc gctgcagcct gagccgccgg cgcccgcccg gccgagcgcc gggggacccc
     1381 gggcccgcga tgtggcccag tacctgctct cagacagcct cttcgtgtgg gttctagtaa
     1441 ataccgcttg ctgtgttttg atgttggtgg ctaagctcat ccagtgtatt gtgtttggcc
     1501 ctcttcgagt gagtgagaga cagcatctca aagacaaatt ttggaatttt attttctaca
     1561 agttcatttt catctttggt gtgctgaatg tccagacagt ggaggaggtg gtcatgtggt
     1621 gcctctggtt tgccggactt gtctttctgc acctgatggt tcagctctgc aaggatcgat
     1681 ttgaatatct ttccttctcg cccaccacgc tgatgagcag ccacggtcga gtcctgtccc
     1741 tgttggttgc catgctgctt tcctgctgtg gactggcggc cgtctgctcc atcaccggct
     1801 acacccacgg aatgcacacc ttggctttca tggctgcaga gtctcttctt gtgacagtga
     1861 ggactgctca tgtgatttta cgatacgtaa ttcacctctg ggacctcaac cacgaaggga
     1921 cgtgggaagg aaaggggacg tatgtctatt acacagactt agtcatggag ctcactctcc
     1981 tgtccctgga cctcatgcac catattcaca tgttgttatt tggcaacatc tggttatcca
     2041 tggccagcct ggtcatcttt atgcagctgc gttacctgtt tcatgaggtg caacgtcgaa
     2101 ttcgtcggca caagaactgt ctacgtgtgg ttggaaacat ggaggccagg tttgcagttg
     2161 caactccaga ggagctggct gtcaacaatg acgactgtgc catctgttgg ggctccatgc
     2221 aggctgcgcg gaaactgccc tgtggacatc ttttccacaa ctcctgtctt cgttcctggc
     2281 tagaacaaga cacctcctgt ccaacatgca gaatgtctct taatattgcc gacaataatc
     2341 gtgtcaggga agaacatcaa ggagagaact tggatgagaa tttggttcct gtagcagcag
     2401 ccgaagggag acctcgctta aaccaacaca atcacttctt ccatttcgat gggtctcgga
     2461 ttgcgagctg gctgccgagt ttttcggttg aagtgatgca caccaccaac attcttggca
     2521 ttacgcaggc cagcaactcc cagctcaatg caatggctca tcagattcaa gagatgtttc
     2581 cccaggtccc ataccatctg gtactgcagg accttcagct gacacgctca gttgaaataa
     2641 caacagacaa tattttagaa ggacggattc aagtaccttt tcctacacag cggtcagata
     2701 gcatcagacc tgcattgaac agtcctgtgg aaaggccaag cagtgaccag gaagagggag
     2761 aaacttctgc tcagaccgag cgtgtgccac tggacctcag tcctcgcctg gaggagacgc
     2821 tggacttcgg cgaggtggaa gtggagccca gtgaggtgga agacttcgag gctcgtggga
     2881 gccgcttctc caagtctgct gatgagagac agcgcatgct ggtgcagcgt aaggacgaac
     2941 tcctccagca agctcgcgca ggtgccgcgg gcggaagttc tgaagatgat gcggcctcag
     3001 agagcttcct cccctcggaa ggtgcgtcct ctgaccccgt gaccctgcgt cgaaggatgc
     3061 tggctgccgc cgcggaacgg aggcttcaga agcagcagac ctcctagcga agccgaattc
     3121 cagcacactg gcggccgtta ctagagtcga cccgggcggc cgcttccctt tagtgagggt
     3181 taatgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     3241 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     3301 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     3361 agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcc
     3421 gataaggatc gatccgggct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     3481 acagttgcgc agcctgaatg gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg
     3541 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3601 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3661 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3721 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3781 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3841 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3901 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     3961 atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgcg
     4021 gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt
     4081 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4141 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4201 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4261 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4321 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4381 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     4441 gcttgattct tctgacacaa cagtctcgaa cttaaggcta gagccaccat gattgaacaa
     4501 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4561 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4621 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4681 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4741 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4801 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4861 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4921 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4981 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5041 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5101 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5161 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5221 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5281 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgat
     5341 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
     5401 atagcgataa ggatccgcgt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     5461 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     5521 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     5581 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     5641 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     5701 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5761 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5821 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5881 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5941 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     6001 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     6061 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     6121 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6181 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     6241 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6301 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6361 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6421 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6481 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6541 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6601 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6661 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6721 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6781 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6841 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6901 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6961 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     7021 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7081 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7141 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7201 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7261 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7321 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7381 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7441 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatggctcg acagatct
//
