LOCUS       sequence-339656-        8559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 3xT7-tagged nuclear form of SREBP1a in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8559)
  AUTHORS   Jang Y, Park YK, Lee JE, Wan D, Tran N, Gavrilova O, Ge K
  TITLE     MED1 is a lipogenesis coactivator required for postnatal adipose 
            expansion.
  JOURNAL   Genes Dev. 2021 Apr 22. pii: gad.347583.120. doi: 
            10.1101/gad.347583.120.
  PUBMED    33888555
REFERENCE   2  (bases 1 to 8559)
  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: "Genes Dev. 
            2021 Apr 22. pii: gad.347583.120. doi: 10.1101/gad.347583.120."
COMMENT     Sequence Label: pMSCVhyg-3xT7-nSREBP1a
FEATURES             Location/Qualifiers
     source          1..8559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             11..527
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    591..932
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             999..1415
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1345..1367
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1383..1399
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1437..1446
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1581..1586,1587..1610,1611..1646,1647..1670)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     promoter        3064..3563
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3099..3122)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3496..3515
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3581..4606
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3581..4606
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     LTR             4964..5478
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5640..5656
                     /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(5664..5670,5671..5688,5689..5694))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5709..5730
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5847..5864)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6018..6606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6098..6117)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6777..7568,7569..7637))
                     /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     7400..7419
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7638..7742)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7810..7828
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7866..7888)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7988..8007
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8201..8223
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8351..8370
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat
       61 tttgcaaggc atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac
      121 agagagacag cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
      181 tcagggccaa gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac
      241 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
      301 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
      361 agcccacaac ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc
      421 gtattcccaa taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg
      481 agggtctcct cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc
      541 gagatttgga gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca
      601 gcggtcgttt cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc
      661 gcctgcgtct gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg
      721 acgagttctg aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt
      781 tttgtggccc gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg
      841 ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg
      901 tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc
      961 tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct
     1021 taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag
     1081 atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg
     1141 gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct
     1201 tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct
     1261 tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc
     1321 ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct
     1381 ccctttatcc agccctcact ccttctctag gcgccggaat tagatctctc gacgaggcca
     1441 ccatggcttc tatgaccggc gggcagcaaa tgggcggctc ttcggccagc atggcttcta
     1501 tgaccggtgg ccaacagatg ggtggttcct cagctagaat ggcgagtatg accggtggcc
     1561 aacagatggg tggtagtgga agcgcgtgga gccaccctca gttcgagaag ggaggaggaa
     1621 gcggcggagg cagcggagga agcgcttgga gccacccgca gttcgagaaa ggagctagat
     1681 cacccgggct cgacatggac gagctggcct tcggtgaggc ggctctggaa cagacactgg
     1741 ccgagatgtg cgaactggac acagcggttt tgaacgacat cgaagacatg ctccagctca
     1801 tcaacaacca agacagtgac ttcccgggcc tgtttgacgc cccctatgct gggggtgaga
     1861 caggggacac aggccccagc agcccaggtg ccaactctcc tgagagcttc tcttctgctt
     1921 ctctggcctc ctctctggaa gccttcctgg gaggacccaa ggtgacacct gcacccttgt
     1981 cccctccacc atcggcaccc gctgctttaa agatgtaccc gtccgtgtcc cccttttccc
     2041 ctgggcctgg gatcaaagag gagccagtgc cactcaccat cctacagcct gcagcgccac
     2101 agccgtcacc ggggaccctc ctgcctccga gcttccccgc accacccgta cagctcagcc
     2161 ctgcgcccgt gctgggttac tcgagcctgc cttcaggctt ctcagggacc cttccaggaa
     2221 acactcagca gccaccatct agcctgccgc tggcccctgc accaggagtc ttgcccaccc
     2281 ctgccctgca cacccaggtc caaagcttgg cctcccagca gccgctgcca gcctcagcag
     2341 cccctagaac aaacactgtg acctcacagg tccagcaggt cccagttgta ctgcagccac
     2401 acttcatcaa ggcagactca ctgctgctga cagctgtgaa gacagatgca ggagccacca
     2461 tgaagactgc aggcatcagc accctggctc ctggcacagc cgtgcaggca ggtcccctgc
     2521 agaccctggt gagtggaggg accatcttgg ccacagtacc tttggttgtg gacacagaca
     2581 aactgcccat ccaccgactc gcagctggca gcaaggccct aggctcagct cagagccgtg
     2641 gtgagaagcg cacagcccac aatgccattg agaagcgcta ccggtcctct atcaatgaca
     2701 agattgtgga gctcaaagac ctggtggtgg gcactgaagc aaagctgaat aaatctgctg
     2761 tcttgcgcaa ggccatcgac tacatccgct tcttgcagca cagcaaccag aagctcaagc
     2821 aggagaacct gaccctacga agtgcacaca aaagcaaatc actgaaggac ctggtgtcag
     2881 cttgtggcag tggaggaggc acagatgtgt ctatggaggg catgaaaccc gaagtggtgg
     2941 agacgcttac ccctccaccc tcagacgccg gctcaccctc ccagagtagc cccttgtctt
     3001 ttggcagcag agctagcagc agtggtggca gtgactctga gtagtaagtt aacgaattct
     3061 accgggtagg ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc
     3121 tgggcacttg gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta
     3181 ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag
     3241 tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag
     3301 cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc
     3361 tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa
     3421 ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt
     3481 cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct
     3541 cttcctcatc tccgggcctt tcgacctgca tcccgccacc atgaaaaagc ctgaactcac
     3601 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
     3661 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
     3721 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
     3781 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct
     3841 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
     3901 actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct
     3961 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
     4021 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     4081 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     4141 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
     4201 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     4261 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     4321 ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct
     4381 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     4441 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     4501 acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga
     4561 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaatagagta gatgccgacc
     4621 gaacaagagc tgatttcgag aacgcctcag ccagcaactc gcgcgagcct agcaaggcaa
     4681 atgcgagaga acggccttac gcttggtggc acagttctcg tccacagttc gctaagctcg
     4741 ctcggctggg tcgcgggagg gccggtcgca gtgattcagg cccttctgga ttgtgttggt
     4801 ccccagggca cgattgtcat gcccacgcac tcgggtgatc tgactgatcc cgcagattgg
     4861 agatcgccgc ccgtgcctgc cgattgggtg cagatccgtc gacctgcagc caagcttatc
     4921 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg
     4981 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa tacataactg
     5041 agaatagaga agttcagatc aaggttagga acagagagac agcagaatat gggccaaaca
     5101 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
     5161 tgcggtcccg ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
     5221 acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt
     5281 cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact cggcgcgcca
     5341 gtcctccgat agactgcgtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg
     5401 catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg
     5461 tcagcggggg tctttcatgg gtaacagttt cttgaagttg gagaacaaca ttctgagggt
     5521 aggagtcgaa tattaagtaa tcctgactca attagccact gttttgaatc cacatactcc
     5581 aatactcctg aaatagttca ttatggacag cgcagaaaga gctggggaga attgtgaaat
     5641 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5701 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5761 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5821 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5881 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5941 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6001 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6061 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6121 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6181 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6241 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6301 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6361 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6421 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6481 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6541 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6601 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6661 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6721 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6781 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6841 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6901 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6961 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7021 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7081 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7141 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7201 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7261 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7321 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7381 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7441 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7501 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7561 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7621 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7681 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7741 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     7801 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     7861 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     7921 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     7981 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     8041 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
     8101 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     8161 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     8221 acgacggcgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     8281 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     8341 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     8401 tgccggccac gatgcgtccg gcgtagaggc gattagtcca atttgttaaa gacaggatat
     8461 cagtggtcca ggctctagtt ttgactcaac aatatcacca gctgaagcct atagagtacg
     8521 agccatagat aaaataaaag attttattta gtctccaga
//
