LOCUS       sequence-409985-        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; A341)
FEATURES             Location/Qualifiers
     source          1..9023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        83..462
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        463..666
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     617..637
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     711..730
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        711..729
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      755..764
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4462..4479)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4468..4692
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4738..5166
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4825..4844)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5035..5056
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5175..5195)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5180..5509
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5360..5495
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5422..5441
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5576..6370
                     /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(5630..5649)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6240..6259
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6544..6665
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6581..6600)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6635..6654
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6714..6730)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6714..6730)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6727..6749)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6738..6754
                     /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(6762..6768,6769..6786,6787..6792))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6807..6828
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6945..6962)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7116..7704)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7196..7215)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7875..8666,8667..8735))
                     /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     8498..8517
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8736..8840)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8915..8934)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
       61 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      121 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      181 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      241 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      301 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      361 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      421 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      481 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      541 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      601 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      661 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
      721 cactataggg agacccaagc tggctagcgg atccgccacc atggcctcta tgaccggcgg
      781 ccagcaaatg ggtatggcct ctatgaccgg cggccagcaa atgggagcat cgatggacga
      841 gccacccttc agcgaggcgg ctttggagca ggcgctgggc gagccgtgcg atctggacgc
      901 ggcgctgctg accgacatcg aaggtgaagt cggcgcgggg aggggtaggg ccaacggcct
      961 ggacgcccca agggcgggcg cagatcgcgg agctatggat tgcactttcg aagacatgct
     1021 tcagcttatc aacaaccaag acagtgactt ccctggccta tttgacccac cctatgctgg
     1081 gagtggggca gggggcacag accctgccag ccccgatacc agctccccag gcagcttgtc
     1141 tccacctcct gccacattga gctcctctct tgaagccttc ctgagcgggc cgcaggcagc
     1201 gccctcaccc ctgtcccctc cccagcctgc acccactcca ttgaagatgt acccgtccat
     1261 gcccgctttc tcccctgggc ctggtatcaa ggaagagtca gtgccactga gcatcctgca
     1321 gacccccacc ccacagcccc tgccaggggc cctcctgcca cagagcttcc cagccccagc
     1381 cccaccgcag ttcagctcca cccctgtgtt aggctacccc agccctccgg gaggcttctc
     1441 tacaggaagc cctcccggga acacccagca gccgctgcct ggcctgccac tggcttcccc
     1501 gccaggggtc ccgcccgtct ccttgcacac ccaggtccag agtgtggtcc cccagcagct
     1561 actgacagtc acagctgccc ccacggcagc ccctgtaacg accactgtga cctcgcagat
     1621 ccagcaggtc ccggtcctgc tgcagcccca cttcatcaag gcagactcgc tgcttctgac
     1681 agccatgaag acagacggag ccactgtgaa ggcggcaggt ctcagtcccc tggtctctgg
     1741 caccactgtg cagacagggc ctttgccgac cctggtgagt gccgcaacca tcttggcaac
     1801 agtcccactg gtcgtagatg cggagaagct gcctatcaac cggctcgcag ctgccagcaa
     1861 ggccccggcc tctgcccaga gccgtggaga gaagcgcaca gcccacaacg ccattgagaa
     1921 gcgctaccgc tcctccatca atgacaaaat cattgagctc aaggatctgg tggtgggcac
     1981 tgaggcaaag ctgaataaat ctgctgtctt gcgcaaggcc atcgactaca ttcgctttct
     2041 gcaacacagc aaccagaagc ttaagcagga gaacctaagt ctgcgcactg ctgtccacaa
     2101 aagcaaatct ctgaaggatc tggtgtcggc ctgtggcagt ggagggaaca cagacgtgct
     2161 catggagggc gtgaagactg aggtggagga cacactgacc ccacccccct cggatgctgg
     2221 ctcacctttc cagagcagcc ccttgtccct tggcagcagg ggcagtggca gcggtggcag
     2281 tggcagtgac tcggagcctg acagcccagt ctttgaggac agcaaggcaa agccagagca
     2341 gcggccgtct ctgcacagcc gcggcatgct ggaccgctcc cgcctggccc tgtgcacgct
     2401 cgtcttcctc tgcctgtcct gcaacccctt ggcctccttg ctgggggccc gggggcttcc
     2461 cagcccctca gataccacca gcgtctacca tagccctggg cgcaacgtgc tgggcaccga
     2521 gtctagagat ggccctggct gggcccagtg gctgctgccc ccagtggtct ggctgctcaa
     2581 tgggctgttg gtgctcgtct ccttggtgct tctctttgtc tacggtgagc cagtcacacg
     2641 gccccactca ggccccgccg tgtacttctg gaggcatcgc aagcaggctg acctggacct
     2701 ggcccgggga gactttgccc aggctgccca gcagctgtgg ctggccctgc gggcactggg
     2761 ccggcccctg cccacctccc acctggacct ggcttgtagc ctcctctgga acctcatccg
     2821 tcacctgctg cagcgtctct gggtgggccg ctggctggca ggccgggcag ggggcctgca
     2881 gcaggactgt gctctgcgag tggatgcaag cgccagcgcc cgagacgcag ccctggtcta
     2941 ccataagctg caccagctgc acaccatggg gaagcacaca ggcgggcacc tcactgccac
     3001 caacctggcg ctgagtgccc tgaacctggc agagtgtgca ggggatgccg tgtctgtggc
     3061 gacgctggcc gagatctatg tggcggctgc attgagagtg aagaccagtc tcccacgggc
     3121 cttgcatttt ctgacacgct tcttcctgag cagtgcccgc caggcctgcc tggcacagag
     3181 tggctcagtg cctcctgcca tgcagtggct ctgccacccc gtgggccacc gtttcttcgt
     3241 ggatggggac tggtccgtgc tcagtacccc atgggagagc ctgtacagct tggccgggaa
     3301 cccagtggac cccctggccc aggtgactca gctattccgg gaacatctct tagagcgagc
     3361 actgaactgt gtgacccagc ccaaccccag ccctgggtca gctgatgggg acaaggaatt
     3421 ttcggatgcc ctcgggtacc tgcagctgct gaacagctgt tctgatgctg cgggggctcc
     3481 tgcctacagc ttctccatca gttccagcat ggccaccacc accggcgtag acccggtggc
     3541 caagtggtgg gcctctctga cagctgtggt gatccactgg ctgcggcggg atgaggaggc
     3601 ggctgagcgg ctgtgcccgc tggtggagca cctgccccgg gtgctgcagg agtctgagag
     3661 acccctgccc agggcagctc tgcactcctt caaggctgcc cgggccctgc tgggctgtgc
     3721 caaggcagag tctggtccag ccagcctgac catctgtgag aaggccagtg ggtacctgca
     3781 ggacagcctg gctaccacac cagccagcag ctccattgac aaggccgtgc agctgttcct
     3841 gtgtgacctg cttcttgtgg tgcgcaccag cctgtggcgg cagcagcagc ccccggcccc
     3901 ggccccagca gcccagggca ccagcagcag gccccaggct tccgcccttg agctgcgtgg
     3961 cttccaacgg gacctgagca gcctgaggcg gctggcacag agcttccggc ccgccatgcg
     4021 gagggtgttc ctacatgagg ccacggcccg gctgatggcg ggggccagcc ccacacggac
     4081 acaccagctc ctcgaccgca gtctgaggcg gcgggcaggc cccggtggca aaggaggcgc
     4141 ggtggcggag ctggagccgc ggcccacgcg gcgggagcac gcggaggcct tgctgctggc
     4201 ctcctgctac ctgccccccg gcttcctgtc ggcgcccggg cagcgcgtgg gcatgctggc
     4261 tgaggcggcg cgcacactag agaaacttgg cgatcgccgg ctgctgcacg actgtcagca
     4321 gatgctcatg cgcctgggcg gtgggaccac tgtcacttcc agcggagaat tctcaggcgc
     4381 ccccgtgccc taccccgacc ccctggagcc ccggtgagcg gccgctcgag tctagtgggc
     4441 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     4501 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     4561 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     4621 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     4681 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     4741 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4801 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4861 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4921 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4981 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     5041 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     5101 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5161 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     5221 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     5281 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     5341 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     5401 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     5461 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     5521 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     5581 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     5641 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     5701 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     5761 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     5821 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     5881 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     5941 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     6001 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     6061 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     6121 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     6181 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     6241 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     6301 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     6361 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     6421 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     6481 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     6541 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     6601 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     6661 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     6721 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     6781 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     6841 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     6901 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     6961 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     7021 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     7081 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     7141 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     7201 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     7261 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     7321 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     7381 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     7441 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     7501 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     7561 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     7621 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     7681 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     7741 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     7801 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     7861 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     7921 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7981 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     8041 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     8101 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     8161 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     8221 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     8281 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     8341 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     8401 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     8461 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     8521 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     8581 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     8641 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     8701 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     8761 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     8821 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     8881 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     8941 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     9001 tttaagctac aacaaggcaa ggc
//
