LOCUS       sequence-409781-        9023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SREBP-1 (Isoform 1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9023)
  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 9023)
  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 (A317-318; L340)
FEATURES             Location/Qualifiers
     source          1..9023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(58..78)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        63..392
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      243..378
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     305..324
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             459..1253
                     /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(513..532)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1123..1142
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1427..1548
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1464..1483)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1518..1537
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1597..1613)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1597..1613)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1610..1632)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1621..1637
                     /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(1645..1651,1652..1669,1670..1675))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1690..1711
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1828..1845)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1999..2587)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2079..2098)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2758..3549,3550..3618))
                     /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     3381..3400
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3619..3723)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3798..3817)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3989..4368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4369..4572
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4523..4543
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4617..4636
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4617..4635
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4661..4670
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(8368..8385)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8374..8598
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(8644..9023,1..49)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8731..8750)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8941..8962
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
       61 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
      121 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
      181 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
      241 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
      301 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
      361 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
      421 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
      481 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
      541 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
      601 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
      661 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
      721 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
      781 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
      841 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
      901 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
      961 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     1021 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     1081 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     1141 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     1201 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     1261 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     1321 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     1381 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     1441 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     1501 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     1561 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     1621 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     1681 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     1741 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     1801 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1861 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1921 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1981 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2041 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2101 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2161 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2221 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2281 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2341 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2401 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2461 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2521 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2581 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2641 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2701 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2761 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2821 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2881 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2941 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3001 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3061 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3121 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3181 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3241 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3301 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3361 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3421 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     3481 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     3541 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     3601 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3661 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     3721 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     3781 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     3841 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     3901 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     3961 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
     4021 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     4081 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     4141 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     4201 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     4261 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     4321 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     4381 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     4441 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     4501 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     4561 ataagcagag ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat
     4621 acgactcact atagggagac ccaagctggc tagcggatcc gccaccatgg cctctatgac
     4681 cggcggccag caaatgggta tggcctctat gaccggcggc cagcaaatgg gagcatcgat
     4741 ggacgagcca cccttcagcg aggcggcttt ggagcaggcg ctgggcgagc cgtgcgatct
     4801 ggacgcggcg ctgctgaccg acatcgaagg tgaagtcggc gcggggaggg gtagggccaa
     4861 cggcctggac gccccaaggg cgggcgcaga tcgcggagct atggattgca ctttcgaaga
     4921 catgcttcag cttatcaaca accaagacag tgacttccct ggcctatttg acccacccta
     4981 tgctgggagt ggggcagggg gcacagaccc tgccagcccc gataccagct ccccaggcag
     5041 cttgtctcca cctcctgcca cattgagctc ctctcttgaa gccttcctga gcgggccgca
     5101 ggcagcgccc tcacccctgt cccctcccca gcctgcaccc actccattga agatgtaccc
     5161 gtccatgccc gctttctccc ctgggcctgg tatcaaggaa gagtcagtgc cactgagcat
     5221 cctgcagacc cccaccccac agcccctgcc aggggccctc ctgccacaga gcttcccagc
     5281 cccagcccca ccgcagttca gctccacccc tgtgttaggc taccccagcc ctccgggagg
     5341 cttctctaca ggaagccctc ccgggaacac ccagcagccg ctgcctggcc tgccactggc
     5401 ttccccgcca ggggtcccgc ccgtctcctt gcacacccag gtccagagtg tggtccccca
     5461 gcagctactg acagtcacag ctgcccccac ggcagcccct gtaacgacca ctgtgacctc
     5521 gcagatccag caggtcccgg tcctgctgca gccccacttc atcaaggcag actcgctgct
     5581 tctgacagcc atgaagacag acggagccac tgtgaaggcg gcaggtctca gtcccctggt
     5641 ctctggcacc actgtgcaga cagggccttt gccgaccctg gtgagtgccg caaccatctt
     5701 ggcaacagtc ccactggtcg tagatgcgga gaagctgcct atcaaccggc tcgcacttgg
     5761 cagcaaggcc ccggcctctg cccagagccg tggagagaag cgcacagccc acaacgccat
     5821 tgagaagcgc taccgctcct ccatcaatga caaaatcatt gagctcaagg atctggtggt
     5881 gggcactgag gcaaagctga ataaatctgc tgtcttgcgc aaggccatcg actacattcg
     5941 ctttctgcaa cacagcaacc agaagcttaa gcaggagaac ctaagtctgc gcactgctgt
     6001 ccacaaaagc aaatctctga aggatctggt gtcggcctgt ggcagtggag ggaacacaga
     6061 cgtgctcatg gagggcgtga agactgaggt ggaggacaca ctgaccccac ccccctcgga
     6121 tgctggctca cctttccaga gcagcccctt gtcccttggc agcaggggca gtggcagcgg
     6181 tggcagtggc agtgactcgg agcctgacag cccagtcttt gaggacagca aggcaaagcc
     6241 agagcagcgg ccgtctctgc acagccgcgg catgctggac cgctcccgcc tggccctgtg
     6301 cacgctcgtc ttcctctgcc tgtcctgcaa ccccttggcc tccttgctgg gggcccgggg
     6361 gcttcccagc ccctcagata ccaccagcgt ctaccatagc cctgggcgca acgtgctggg
     6421 caccgagtct agagatggcc ctggctgggc ccagtggctg ctgcccccag tggtctggct
     6481 gctcaatggg ctgttggtgc tcgtctcctt ggtgcttctc tttgtctacg gtgagccagt
     6541 cacacggccc cactcaggcc ccgccgtgta cttctggagg catcgcaagc aggctgacct
     6601 ggacctggcc cggggagact ttgcccaggc tgcccagcag ctgtggctgg ccctgcgggc
     6661 actgggccgg cccctgccca cctcccacct ggacctggct tgtagcctcc tctggaacct
     6721 catccgtcac ctgctgcagc gtctctgggt gggccgctgg ctggcaggcc gggcaggggg
     6781 cctgcagcag gactgtgctc tgcgagtgga tgcaagcgcc agcgcccgag acgcagccct
     6841 ggtctaccat aagctgcacc agctgcacac catggggaag cacacaggcg ggcacctcac
     6901 tgccaccaac ctggcgctga gtgccctgaa cctggcagag tgtgcagggg atgccgtgtc
     6961 tgtggcgacg ctggccgaga tctatgtggc ggctgcattg agagtgaaga ccagtctccc
     7021 acgggccttg cattttctga cacgcttctt cctgagcagt gcccgccagg cctgcctggc
     7081 acagagtggc tcagtgcctc ctgccatgca gtggctctgc caccccgtgg gccaccgttt
     7141 cttcgtggat ggggactggt ccgtgctcag taccccatgg gagagcctgt acagcttggc
     7201 cgggaaccca gtggaccccc tggcccaggt gactcagcta ttccgggaac atctcttaga
     7261 gcgagcactg aactgtgtga cccagcccaa ccccagccct gggtcagctg atggggacaa
     7321 ggaattttcg gatgccctcg ggtacctgca gctgctgaac agctgttctg atgctgcggg
     7381 ggctcctgcc tacagcttct ccatcagttc cagcatggcc accaccaccg gcgtagaccc
     7441 ggtggccaag tggtgggcct ctctgacagc tgtggtgatc cactggctgc ggcgggatga
     7501 ggaggcggct gagcggctgt gcccgctggt ggagcacctg ccccgggtgc tgcaggagtc
     7561 tgagagaccc ctgcccaggg cagctctgca ctccttcaag gctgcccggg ccctgctggg
     7621 ctgtgccaag gcagagtctg gtccagccag cctgaccatc tgtgagaagg ccagtgggta
     7681 cctgcaggac agcctggcta ccacaccagc cagcagctcc attgacaagg ccgtgcagct
     7741 gttcctgtgt gacctgcttc ttgtggtgcg caccagcctg tggcggcagc agcagccccc
     7801 ggccccggcc ccagcagccc agggcaccag cagcaggccc caggcttccg cccttgagct
     7861 gcgtggcttc caacgggacc tgagcagcct gaggcggctg gcacagagct tccggcccgc
     7921 catgcggagg gtgttcctac atgaggccac ggcccggctg atggcggggg ccagccccac
     7981 acggacacac cagctcctcg accgcagtct gaggcggcgg gcaggccccg gtggcaaagg
     8041 aggcgcggtg gcggagctgg agccgcggcc cacgcggcgg gagcacgcgg aggccttgct
     8101 gctggcctcc tgctacctgc cccccggctt cctgtcggcg cccgggcagc gcgtgggcat
     8161 gctggctgag gcggcgcgca cactagagaa acttggcgat cgccggctgc tgcacgactg
     8221 tcagcagatg ctcatgcgcc tgggcggtgg gaccactgtc acttccagcg gagaattctc
     8281 aggcgccccc gtgccctacc ccgaccccct ggagccccgg tgagcggccg ctcgagtcta
     8341 gtgggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     8401 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     8461 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     8521 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     8581 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     8641 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     8701 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     8761 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     8821 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     8881 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     8941 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     9001 tataagggat tttgccgatt tcg
//
