LOCUS       sequence-384520-        6049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Ribophorin I .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6049)
  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 6049)
  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: pcDNA4-RI1-332-MYC-His6
FEATURES             Location/Qualifiers
     source          1..6049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(108..128)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        113..442
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      293..428
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     355..374
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        490..537
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             556..930
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    1060..1181
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1097..1116)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1151..1170
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1230..1246)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1230..1246)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1243..1265)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1254..1270
                     /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(1278..1284,1285..1302,1303..1308))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1323..1344
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1461..1478)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1632..2220)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1712..1731)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2391..3182,3183..3251))
                     /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     3014..3033
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3252..3356)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3431..3450)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3622..4001
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4002..4205
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4156..4176
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4250..4269
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4250..4268
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4342..4351
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     RBS             5258..5266
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             5359..5388
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5404..5421
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5444..5461)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5450..5674
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(5720..6049,1..99)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5807..5826)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6017..6038
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
       61 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
      121 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
      181 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
      241 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
      301 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
      361 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
      421 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
      481 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
      541 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
      601 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
      661 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
      721 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
      781 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
      841 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
      901 gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc
      961 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     1021 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     1081 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     1141 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     1201 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     1261 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     1321 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     1381 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     1441 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     1501 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     1561 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1621 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1681 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1741 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1801 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1861 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1921 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1981 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2041 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     2101 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2161 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2221 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2281 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     2341 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     2401 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     2461 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     2521 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     2581 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     2641 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     2701 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     2761 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     2821 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     2881 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     2941 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     3001 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     3061 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     3121 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     3181 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     3241 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     3301 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     3361 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     3421 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     3481 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     3541 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     3601 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
     3661 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     3721 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     3781 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     3841 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     3901 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     3961 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     4021 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     4081 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     4141 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     4201 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
     4261 actataggga gacccaagct ggctagttaa gcttggtacc gagctcggat ccactagtcc
     4321 agtgtggtgg aattcggatc cgccaccatg gaggcgccag ccgccggctt gtttctgctc
     4381 ctgttgcttg ggacttgggc cccggcgccg ggcagcgcct cctccgaggc accgccgctg
     4441 atcaatgagg acgtgaagcg cacagtggac ctaagcagcc acctggctaa ggtgacggcc
     4501 gaggtggtcc tggcgcacct gggcggcggc tccacgtccc gagctacctc tttcctgctg
     4561 gctttggagc ctgagctcga ggcccggctg gcgcacctgg gcgtgcaggt aaagggagaa
     4621 gatgaggaag agaacaattt ggaagtacgt gaaaccaaaa ttaagggtaa aagtgggaga
     4681 ttcttcacag tcaagctccc agttgctctt gatcctgggg ccaagatttc agtcattgtg
     4741 gaaacagtct acacccatgt gcttcatcca tatccaaccc agatcaccca gtcagagaaa
     4801 cagtttgtgg tgtttgaggg gaaccattat ttctactctc cctatccaac gaagacacaa
     4861 accatgcgtg tgaagcttgc ctctcgaaat gtggagagct acaccaagct ggggaacccc
     4921 acgcgctctg aggacctact ggattatggg cctttcagag atgtgcctgc ctatagtcag
     4981 gatactttta aagtacatta tgagaacaac agccctttcc tgaccatcac cagcatgacc
     5041 cgagtcattg aagtctctca ctggggtaat attgctgtgg aagaaaatgt ggacttaaag
     5101 cacacaggag ctgtgcttaa ggggcctttc tcacgctatg attaccagag acagccagat
     5161 agtggaatat cctccatccg ttcttttaag accatccttc ctgctgctgc ccaggatgtt
     5221 tattaccggg atgagattgg caatgtttct accagccacc tccttatttt ggatgactct
     5281 gtagagatgg aaatccggcc tcgcttccct ctctttggcg ggtggaagac ccattacatc
     5341 gtttctagag ggcccttcga acaaaaactc atctcagaag aggatctgaa tatgcatacc
     5401 ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac tgtgccttct
     5461 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     5521 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     5581 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     5641 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     5701 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     5761 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     5821 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     5881 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     5941 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     6001 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactca
//
