LOCUS       sequence-421642-        9017 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SREBP-1 (isoform 1) with mutations in 
            residues reported to be critical for S1P and S2P cleavage..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9017)
  AUTHORS   Yang M, Mariano J, Su R, Smith CE, Das S, Gill C, Andresson T, 
            Loncarek J, Tsai YC, Weissman AM
  TITLE     SARS-CoV-2 papain-like protease plays multiple roles in regulating 
            cellular proteins in the endoplasmic reticulum.
  JOURNAL   J Biol Chem. 2023 Oct 12;299(12):105346. doi: 
            10.1016/j.jbc.2023.105346.
  PUBMED    37838170
REFERENCE   2  (bases 1 to 9017)
  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: "J Biol 
            Chem."; date: "2023-10-12"; volume: "299(12):105346. doi"; pages: " 
            10.1016/j.jbc.2023.105346"
COMMENT     Sequence Label: pCDNA3-SREBP-1 S1P and S2P mutations (U124AFG2...
FEATURES             Location/Qualifiers
     source          1..9017
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     393..412
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(631..735)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(810..829)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1001..1380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1381..1584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1535..1555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1629..1648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1629..1647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1673..1682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5374..5391)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5380..5604
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5650..6078
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5737..5756)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5947..5968
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6087..6107)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6092..6421
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6272..6407
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6334..6353
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6488..7282
                     /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(6542..6561)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7152..7171
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7456..7577
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7493..7512)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7547..7566
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7626..7642)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7626..7642)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7639..7661)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7650..7666
                     /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(7674..7680,7681..7698,7699..7704))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7719..7740
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7857..7874)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8028..8616)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8108..8127)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8787..9017,1..561,562..630))
                     /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"
ORIGIN
        1 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
       61 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
      121 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
      181 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
      241 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
      301 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
      361 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
      421 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
      481 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
      541 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
      601 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
      661 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
      721 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
      781 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
      841 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
      901 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
      961 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
     1021 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     1081 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     1141 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     1201 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     1261 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     1321 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     1381 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     1441 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     1501 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     1561 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
     1621 atcgaaatta atacgactca ctatagggag acccaagctg gctagcggat ccgccaccat
     1681 ggcctctatg accggcggcc agcaaatggg tatggcctct atgaccggcg gccagcaaat
     1741 gggagcatcg atggacgagc cacccttcag cgaggcggct ttggagcagg cgctgggcga
     1801 gccgtgcgat ctggacgcgg cgctgctgac cgacatcgaa ggtgaagtcg gcgcggggag
     1861 gggtagggcc aacggcctgg acgccccaag ggcgggcgca gatcgcggag ctatggattg
     1921 cactttcgaa gacatgcttc agcttatcaa caaccaagac agtgacttcc ctggcctatt
     1981 tgacccaccc tatgctggga gtggggcagg gggcacagac cctgccagcc ccgataccag
     2041 ctccccaggc agcttgtctc cacctcctgc cacattgagc tcctctcttg aagccttcct
     2101 gagcgggccg caggcagcgc cctcacccct gtcccctccc cagcctgcac ccactccatt
     2161 gaagatgtac ccgtccatgc ccgctttctc ccctgggcct ggtatcaagg aagagtcagt
     2221 gccactgagc atcctgcaga cccccacccc acagcccctg ccaggggccc tcctgccaca
     2281 gagcttccca gccccagccc caccgcagtt cagctccacc cctgtgttag gctaccccag
     2341 ccctccggga ggcttctcta caggaagccc tcccgggaac acccagcagc cgctgcctgg
     2401 cctgccactg gcttccccgc caggggtccc gcccgtctcc ttgcacaccc aggtccagag
     2461 tgtggtcccc cagcagctac tgacagtcac agctgccccc acggcagccc ctgtaacgac
     2521 cactgtgacc tcgcagatcc agcaggtccc ggtcctgctg cagccccact tcatcaaggc
     2581 agactcgctg cttctgacag ccatgaagac agacggagcc actgtgaagg cggcaggtct
     2641 cagtcccctg gtctctggca ccactgtgca gacagggcct ttgccgaccc tggtgagtgg
     2701 cggaaccatc ttggcaacag tcccactggt cgtagatgcg gagaagctgc ctatcaaccg
     2761 gctcgcagct ggcagcaagg ccccggcctc tgcccagagc cgtggagaga agcgcacagc
     2821 ccacaacgcc attgagaagc gctaccgctc ctccatcaat gacaaaatca ttgagctcaa
     2881 ggatctggtg gtgggcactg aggcaaagct gaataaatct gctgtcttgc gcaaggccat
     2941 cgactacatt cgctttctgc aacacagcaa ccagaagctt aagcaggaga acctaagtct
     3001 gcgcactgct gtccacaaaa gcaaatctct gaaggatctg gtgtcggcct gtggcagtgg
     3061 agggaacaca gacgtgctca tggagggcgt gaagactgag gtggaggaca cactgacccc
     3121 acccccctcg gatgctggct cacctttcca gagcagcccc ttgtcccttg gcagcagggg
     3181 cagtggcagc ggtggcagtg gcagtgactc ggagcctgac agcccagtct ttgaggacag
     3241 caaggcaaag ccagagcagc ggccgtctct gcacagccgg ggcatgctgg catccctggc
     3301 cctgtgcacg ctcgtcttcc tctgcctgtc ctgcaacccc ttggcctcct tgctgggggc
     3361 ccgggggctt cccagcccct cagataccac cagcgtctac catagccctg gggccaacgt
     3421 gctgggcacc gagtctagag atggccctgg ctgggcccag tggctgctgc ccccagtggt
     3481 ctggctgctc aatgggctgt tggtgctcgt ctccttggtg cttctctttg tctacggtga
     3541 gccagtcaca cggccccact caggccccgc cgtgtacttc tggaggcatc gcaagcaggc
     3601 tgacctggac ctggcccggg gagactttgc ccaggctgcc cagcagctgt ggctggccct
     3661 gcgggcactg ggccggcccc tgcccacctc ccacctggac ctggcttgta gcctcctctg
     3721 gaacctcatc cgtcacctgc tgcagcgtct ctgggtgggc cgctggctgg caggccgggc
     3781 agggggcctg cagcaggact gtgctctgcg agtggatgca agcgccagcg cccgagacgc
     3841 agccctggtc taccataagc tgcaccagct gcacaccatg gggaagcaca caggcgggca
     3901 cctcactgcc accaacctgg cgctgagtgc cctgaacctg gcagagtgtg caggggatgc
     3961 cgtgtctgtg gcgacgctgg ccgagatcta tgtggcggct gcattgagag tgaagaccag
     4021 tctcccacgg gccttgcatt ttctgacacg cttcttcctg agcagtgccc gccaggcctg
     4081 cctggcacag agtggctcag tgcctcctgc catgcagtgg ctctgccacc ccgtgggcca
     4141 ccgtttcttc gtggatgggg actggtccgt gctcagtacc ccatgggaga gcctgtacag
     4201 cttggccggg aacccagtgg accccctggc ccaggtgact cagctattcc gggaacatct
     4261 cttagagcga gcactgaact gtgtgaccca gcccaacccc agccctgggt cagctgatgg
     4321 ggacaaggaa ttttcggatg ccctcgggta cctgcagctg ctgaacagct gttctgatgc
     4381 tgcgggggct cctgcctaca gcttctccat cagttccagc atggccacca ccaccggcgt
     4441 agacccggtg gccaagtggt gggcctctct gacagctgtg gtgatccact ggctgcggcg
     4501 ggatgaggag gcggctgagc ggctgtgccc gctggtggag cacctgcccc gggtgctgca
     4561 ggagtctgag agacccctgc ccagggcagc tctgcactcc ttcaaggctg cccgggccct
     4621 gctgggctgt gccaaggcag agtctggtcc agccagcctg accatctgtg agaaggccag
     4681 tgggtacctg caggacagcc tggctaccac accagccagc agctccattg acaaggccgt
     4741 gcagctgttc ctgtgtgacc tgcttcttgt ggtgcgcacc agcctgtggc ggcagcagca
     4801 gcccccggcc ccggccccag cagcccaggg caccagcagc aggccccagg cttccgccct
     4861 tgagctgcgt ggcttccaac gggacctgag cagcctgagg cggctggcac agagcttccg
     4921 gcccgccatg cggagggtgt tcctacatga ggccacggcc cggctgatgg cgggggccag
     4981 ccccacacgg acacaccagc tcctcgaccg cagtctgagg cggcgggcag gccccggtgg
     5041 caaaggaggc gcggtggcgg agctggagcc gcggcccacg cggcgggagc acgcggaggc
     5101 cttgctgctg gcctcctgct acctgccccc cggcttcctg tcggcgcccg ggcagcgcgt
     5161 gggcatgctg gctgaggcgg cgcgcacact agagaaactt ggcgatcgcc ggctgctgca
     5221 cgactgtcag cagatgctca tgcgcctggg cggtgggacc actgtcactt ccagcggaga
     5281 attctcaggc gcccccgtgc cctaccccga ccccctggag ccccggtgag cggccgctcg
     5341 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     5401 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     5461 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     5521 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     5581 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     5641 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     5701 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     5761 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5821 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     5881 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5941 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     6001 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     6061 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     6121 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     6181 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     6241 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     6301 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     6361 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     6421 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     6481 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     6541 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     6601 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     6661 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     6721 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     6781 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     6841 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     6901 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     6961 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     7021 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     7081 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     7141 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     7201 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     7261 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     7321 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     7381 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     7441 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     7501 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     7561 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     7621 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     7681 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     7741 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     7801 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     7861 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     7921 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     7981 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     8041 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     8101 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     8161 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     8221 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     8281 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     8341 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     8401 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     8461 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     8521 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     8581 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     8641 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     8701 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     8761 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     8821 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     8881 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     8941 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     9001 cctgcaactt tatccgc
//
