LOCUS       sequence-275428-        8483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Flag-tagged SREBP1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8483)
  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 8483)
  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-SREBP1
FEATURES             Location/Qualifiers
     source          1..8483
                     /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(4439..4456)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4445..4669
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4715..5143
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4802..4821)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5012..5033
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5152..5172)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5157..5486
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5337..5472
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5399..5418
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5534..5581
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5600..5998
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    6156..6277
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6193..6212)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6247..6266
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6326..6342)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6326..6342)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6339..6361)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6350..6366
                     /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(6374..6380,6381..6398,6399..6404))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6419..6440
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6557..6574)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6728..7316)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6808..6827)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7487..8278,8279..8347))
                     /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     8110..8129
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8348..8452)
                     /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 catggacgag ctggccttcg gtgaggcggc tctggaacag acactggccg
     1021 agatgtgcga actggacaca gcggttttga acgacatcga agacatgctc cagctcatca
     1081 acaaccaaga cagtgacttc ccgggcctgt ttgacgcccc ctatgctggg ggtgagacag
     1141 gggacacagg ccccagcagc ccaggtgcca actctcctga gagcttctct tctgcttctc
     1201 tggcctcctc tctggaagcc ttcctgggag gacccaaggt gacacctgca cccttgtccc
     1261 ctccaccatc ggcacccgct gctttaaaga tgtacccgtc cgtgtccccc ttttcccctg
     1321 ggcctgggat caaagaggag ccagtgccac tcaccatcct acagcctgca gcgccacagc
     1381 cgtcaccggg gaccctcctg cctccgagct tccccgcacc acccgtacag ctcagccctg
     1441 cgcccgtgct gggttactcg agcctgcctt caggcttctc agggaccctt ccaggaaaca
     1501 ctcagcagcc accatctagc ctgccgctgg cccctgcacc aggagtcttg cccacccctg
     1561 ccctgcacac ccaggtccaa agcttggcct cccagcagcc gctgccagcc tcagcagccc
     1621 ctagaacaaa cactgtgacc tcacaggtcc agcaggtccc agttgtactg cagccacact
     1681 tcatcaaggc agactcactg ctgctgacag ctgtgaagac agatgcagga gccaccatga
     1741 agactgcagg catcagcacc ctggctcctg gcacagccgt gcaggcaggt cccctgcaga
     1801 ccctggtgag tggagggacc atcttggcca cagtaccttt ggttgtggac acagacaaac
     1861 tgcccatcca ccgactcgca gctggcagca aggccctagg ctcagctcag agccgtggtg
     1921 agaagcgcac agcccacaat gccattgaga agcgctaccg gtcctctatc aatgacaaga
     1981 ttgtggagct caaagacctg gtggtgggca ctgaagcaaa gctgaataaa tctgctgtct
     2041 tgcgcaaggc catcgactac atccgcttct tgcagcacag caaccagaag ctcaagcagg
     2101 agaacctgac cctacgaagt gcacacaaaa gcaaatcact gaaggacctg gtgtcagctt
     2161 gtggcagtgg aggaggcaca gatgtgtcta tggagggcat gaaacccgaa gtggtggaga
     2221 cgcttacccc tccaccctca gacgccggct caccctccca gagtagcccc ttgtcttttg
     2281 gcagcagagc tagcagcagt ggtggtagtg actctgagcc cgacagtcca gcctttgagg
     2341 atagccaggt caaagcccag cggctgcctt cacacagccg aggcatgctg gaccgctccc
     2401 gcctggccct gtgtgtactg gcctttctgt gtctgacctg caatcctttg gcctcgcttt
     2461 tcggctgggg cattctcact ccctctgatg ctacgggtac acaccgtagt tctgggcgca
     2521 gcatgctgga ggcagagagc agagatggct ctaattggac ccagtggttg ctgccacccc
     2581 tagtctggct ggccaatgga ctactagtgt tggcctgctt ggctcttctc tttgtctatg
     2641 gggaacctgt gactaggcca cactctggcc cggctgtaca cttctggaga catcgcaaac
     2701 aagctgacct ggatttggcc cggggagatt tcccccaggc tgctcaacag ctgtggctgg
     2761 ccctgcaagc gctgggccgg cccctgccca cctcaaacct ggatctggcc tgcagtctgc
     2821 tttggaacct catccgccac ctgctccagc gtctctgggt gggccgctgg ctggcaggcc
     2881 aggccggggg cctgctgagg gaccgtgggc tgaggaagga tgcccgtgcc agtgcccggg
     2941 atgcggctgt tgtctaccat aagctgcacc agctgcatgc catgggcaag tacacaggag
     3001 gacatcttgc tgcttctaac ctggcactaa gtgccctcaa cctggctgag tgcgcaggag
     3061 atgctatctc catggcaaca ctggcagaga tctatgtggc agcggccctg agggtcaaaa
     3121 ccagcctccc aagagccctg cacttcttga cacgtttctt cctgagcagc gcccgccagg
     3181 cctgcctagc acagagcggc tcggtgcctc ttgccatgca gtggctctgc caccctgtag
     3241 gtcaccgttt ctttgtggac ggggactggg ccgtgcacgg tgcccccccg gagagcctgt
     3301 acagcgtggc tgggaaccca gtggatccgc tggcccaggt gacccggcta ttccgtgaac
     3361 atctcctaga gcgagcgttg aactgtattg ctcagcccag cccaggggca gctgacggag
     3421 acagggagtt ctcagatgcc cttggatatc tgcagttgct aaatagctgt tctgatgctg
     3481 ccggggctcc tgcttgcagt ttctctgtca gctccagcat ggctgccacc actggcccag
     3541 acccagtggc caagtggtgg gcctcactga cagctgtggt gatccactgg ctgaggcggg
     3601 atgaagaggc agctgagcgc ttgtacccac tggtagagca tatcccccag gtgctgcagg
     3661 acactgagag acccctgccc agggcagctc tgtactcctt caaggctgcc cgggctctgc
     3721 tggaccacag aaaggtggaa tctagcccag ccagcctggc catctgtgag aaggccagtg
     3781 ggtacctgcg ggacagctta gcctctacac caactggcag ttccattgac aaggccatgc
     3841 agctgctcct gtgtgatcta cttcttgtgg cccgtaccag tctgtggcag cggcagcagt
     3901 caccagcttc agtccaggta gctcacggta ccagcaatgg accccaggcc tctgctctgg
     3961 agctgcgtgg tttccaacat gacctgagca gcctgcggcg gttggcacag agcttccggc
     4021 ctgctatgag gagggtattc ctacatgagg ccacagctcg gctgatggca ggagcaagtc
     4081 ctgcccggac acaccagctc ctggatcgca gtctgaggag gagggcaggt tccagtggca
     4141 aaggaggcac tacagctgag ctggagccac ggcccacatg gcgggagcac accgaggccc
     4201 tgctgttggc atcctgctat ctgccccctg ccttcctgtc ggctcctggg cagcgaatga
     4261 gcatgctggc cgaggcggca cgcaccgtag agaagcttgg cgatcaccgg ctactgctgg
     4321 actgccagca gatgctcctg cgcctgggcg gcggaaccac cgtcacttcc agctagctcg
     4381 agccatggaa gcttgatatc taactgactg aaccggtggt accaaacccg ctgatcagcc
     4441 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     4501 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     4561 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     4621 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     4681 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     4741 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4801 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4861 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4921 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4981 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     5041 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     5101 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     5161 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5221 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5281 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5341 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5401 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     5461 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     5521 ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
     5581 caaggtgagg aactaaacca tggccaagcc tttgtctcaa gaagaatcca ccctcattga
     5641 aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc
     5701 agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg
     5761 accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac
     5821 ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg
     5881 acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca
     5941 gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaagc
     6001 acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct
     6061 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     6121 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     6181 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6241 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     6301 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6361 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6421 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6481 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6541 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6601 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6661 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6721 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6781 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6841 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6901 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6961 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7021 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7081 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7141 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     7201 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7261 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7321 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7381 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7441 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7501 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7561 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7621 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7681 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7741 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7801 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7861 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7921 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7981 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8041 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8101 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8161 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8221 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8281 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8341 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8401 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8461 cccgaaaagt gccacctgac gtc
//
