LOCUS       sequence-275429-        8471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Flag-tagged SREBP2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8471)
  AUTHORS   Zhang D, Tomisato W, Su L, Sun L, Choi JH, Zhang Z, Wang KW, Zhan X,
            Choi M, Li X, Tang M, Castro-Perez JM, Hildebrand S, Murray AR, 
            Moresco EMY, Beutler B
  TITLE     Skin-specific regulation of SREBP processing and lipid biosynthesis 
            by glycerol kinase 5.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Jun 27;114(26):E5197-E5206. doi: 
            10.1073/pnas.1705312114. Epub 2017 Jun 12.
  PUBMED    28607088
REFERENCE   2  (bases 1 to 8471)
  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"; doi: 
            "10.1073/pnas.1705312114"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2017-06-27- 27"; volume: "114"; issue: "26"; pages: 
            "E5197-E5206"
COMMENT     Sequence Label: pcDNA6-N-Flag-SREBP2
FEATURES             Location/Qualifiers
     source          1..8471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      921..930
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             930..953
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4427..4444)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4433..4657
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4703..5131
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4790..4809)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5000..5021
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5140..5160)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5145..5474
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5325..5460
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5387..5406
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5522..5569
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5588..5986
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    6144..6265
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6181..6200)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6235..6254
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6314..6330)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6314..6330)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6327..6349)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6338..6354
                     /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(6362..6368,6369..6386,6387..6392))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6407..6428
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6545..6562)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6716..7304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6796..6815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7475..8266,8267..8335))
                     /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     8098..8117
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8336..8440)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 tagagctcct taaggttaac gccaccatgg actacaaaga cgatgacgac aagtctagag
      961 aattcggatc catggatgag agcagcgagc tgggcgttct ggagaccatg gagaccctca
     1021 cggagctggg cgatgagctg actctcgggg acatcgacga gatgctacag tttgtcagca
     1081 atcaagtggg agagttccct gatttgtttt cagagcaact ttgtagctcc ttccctggag
     1141 gtggcagcaa tggtggcagt ggcaacaaca gcagtggcag aggcaacaat ggtggagcca
     1201 cagaccctgc agtacagcgg tcattcagcc aggtcccatt atctaccttc tctccctcag
     1261 ctgcctcccc tcaggctccg gccctgcagg tcaaagtctc tcctacccca ccaagggcaa
     1321 cccctgttct ccagccgcga ccccagcccc agccccaacc tccagctcag ctgcagcagc
     1381 agacggtgat gatcaccccg acgttcagca ccgctccgca gacgaggatc atccagcagc
     1441 ctttgatata ccagaacgca gcgaccagct ttcaagtcct gcagcctcaa gtgcaaagcc
     1501 tcgtgacgtc cccgcaggtg cagcccgtca ccatccagca gcaggtgcag acggtacagg
     1561 cccagcgggt gttgacgcag acagccaatg gcacactgca gacccttgcc cccgccacgg
     1621 tacagacggt cgctgcgcca caggtgcagc aggtcccggt cctggtgcaa cctcagatca
     1681 ttaagacaga ttcccttgtg ttgaccacgc tgaagacaga tggcagccct gtcatggctg
     1741 cagtccagaa cccggccctc acggccctca ctgcccccat ccagacagcc gcccttcaag
     1801 tgccaaccct ggtgggcagc aacgggacca ttctgaccac aatgcctgtg atgatggggc
     1861 aagagaaagt tcctatcaag caagtgcctg gcggcgtgaa gcagctggat cctcccaaag
     1921 aaggagagag gcggacaaca cacaatatca ttgaaaagcg ctaccggtcc tccatcaacg
     1981 acaaaatcat agagttgaag gacttagtca tggggacaga tgccaagatg cacaagtctg
     2041 gcgttctgag gaaggccatt gattacatca aatatctgca gcaggtcaat cacaagctgc
     2101 gccaggagaa catggtgctg aagctggcca atcagaaaaa caagctcctg aagggcatcg
     2161 acctgggcag tctggtggac agtgatgtgg acttgaaaat tgatgacttt aaccagaatg
     2221 tccttctgat gtctccgccg gcctccgact ccgggtccca ggccggcttc tctccctatt
     2281 ccattgactc tgagccgggc agccctctgc tggatgacgc aaaggtcaag gatgaaccgg
     2341 actctcctcc tgtggcactg ggcatggtgg accgctctcg aatcctctta tgtgtcctca
     2401 ccttcctggg cctctccttt aaccccttga cttccttgct gcagtgggga ggggcccaca
     2461 acactgacca gcacccatac tcaggctcgg gccgcagtgt actgtcactg gagtcaggtg
     2521 ctgggggctg gtttgactgg atggttccaa ctctcctcct gtggctggta aatggtgtga
     2581 ttgtcttgag cgtctttgtg aagctgttgg tccacgggga gccggtgatt cgcccacact
     2641 cacgcccctc agtcaccttc tggagacacc ggaagcaggc agacctagac ctcgccaaag
     2701 gtgatttcgc agctgctgct gccaacctac aaacctgctt atccgtgctg ggccgggcgc
     2761 tgcccacctc ccgcctggac ttggcctgca gcctctcctg gaatgtgatc cgctacagcc
     2821 tgcagaagct gcgcctggta cgctggttac tcaagaaggt cttccagcgc tggcgggcta
     2881 ctcccgccac tgcagctggc tttgaggatg aggctaagag cagcgcaagg gatgcggcac
     2941 tggcctacca caggctgcac cagctgcaca tcacagggaa gcttccggcg ggatctgcct
     3001 gttcagatgt gcacatggct ctgtgtgctg tgaacctggc cgagtgtgcg gaggagaaaa
     3061 tcctaccaag cacactgatt gagatccacc tgacagcggc catggggctc aagacccgct
     3121 gtggaggcaa gctgggcttc ttggctagct acttcttaaa tcgtgcccag agtctgtgtg
     3181 gcccagagca tagcactgtc cccgactccc tgcgctggct atgccatccc ttgggccaga
     3241 agtttttcat ggagcgaagc tggtcgatca agtcagcagc caaggagagc ctgtactgcg
     3301 cccagaggag cccagctgac cccattgccc aggtacacca ggcgttctgc aaaaacctgc
     3361 tagaaagagc ggtggagtcc ttggtgaaac ctcaggccaa gaagaaggca ggcgaccagg
     3421 aagaagagag ctgtgaattt tccagtgctc tagagtattt gaaattactt cattcttttg
     3481 tggactctgt gggatttgtg acctcaccat tctccagcag ttccgtgctc aggtcggccc
     3541 ttggccccga tgtcatctgc cggtggtgga cgtccgcagt caccatggcc atcagctggc
     3601 tccagggaga cgatgcagct gtgcgctctc gttttactga agtagagcgg gtccccaagg
     3661 ccctggaagt gaccgagagt cccttggtga aggccgtctt ctatacctgt agagccatgc
     3721 atgcctccct ctctgggaaa gctgacgggc agcagaactc cttctgtcac tgtgagaggg
     3781 ccagtggtca cctgtggagc agtctcaacg tcagcggcac tacttctgat ccttcactta
     3841 accatgtgat ccagctgttc acctgtgacc tgctactgtc gctgaggaca gcactctggc
     3901 aaaagcaggc cagtgccagc cagctcctgg gtgagaccta ccatgcatcc ggcactgagc
     3961 tggccggctt ccagcgggac ctgggcagcc ttcgaaggct ggcccatagc ttccgcccag
     4021 cctaccgcaa ggtgttcctg catgaagcca ctgtgcgcct gatggcagga gccagcccta
     4081 cccgtacaca ccagctgctg gagcatagcc tacggcggcg ccccactcag aacaccaagc
     4141 atggagaggt agacacctgg cctggccaac gagagcgagc tactgccatc ctgctggcct
     4201 gccgccatct tcccctctct ttcctgtctt ccccgggcca gcgggcagtg ctgctggctg
     4261 aagctgcccg caccctggag aaggtgggtg accgccgttc ctgcagtgac tgccagcaga
     4321 tgattgtcaa gctgggcggt ggcactgcca tcgctgcctc ctgactcgag ccatggaagc
     4381 ttgatatcta actgactgaa ccggtggtac caaacccgct gatcagcctc gactgtgcct
     4441 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     4501 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     4561 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     4621 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     4681 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4741 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4801 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4861 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4921 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4981 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     5041 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     5101 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     5161 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5221 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5281 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     5341 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     5401 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     5461 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg
     5521 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     5581 ctaaaccatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc
     5641 tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag
     5701 cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga
     5761 actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc
     5821 gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct
     5881 cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt
     5941 tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaagcac ttcgtggccg
     6001 aggagcagga ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt
     6061 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     6121 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     6181 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6241 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     6301 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     6361 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6421 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6481 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     6541 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6601 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6661 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6721 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6781 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6841 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6901 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6961 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     7021 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     7081 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7141 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7201 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     7261 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     7321 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     7381 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     7441 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     7501 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     7561 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     7621 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7681 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7741 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7801 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7861 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7921 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7981 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     8041 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     8101 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     8161 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     8221 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     8281 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     8341 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     8401 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     8461 cacctgacgt c
//
