LOCUS       sequence-339654-        7212 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 3xT7-tagged target in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7212)
  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 7212)
  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
FEATURES             Location/Qualifiers
     source          1..7212
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             989..1405
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1335..1357
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1373..1389
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1427..1436
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1571..1576,1577..1600,1601..1636,1637..1660)
                     /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        1707..2206
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1742..1765)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2139..2158
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2224..3249
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2224..3249
                     /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             3607..4121
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4283..4299
                     /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(4307..4313,4314..4331,4332..4337))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4352..4373
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4490..4507)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4661..5249)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4741..4760)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5420..6211,6212..6280))
                     /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     6043..6062
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6281..6385)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6453..6471
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6509..6531)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6631..6650
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6844..6866
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6994..7013
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tagatctctc gacgaggcca ccatggcttc
     1441 tatgaccggc gggcagcaaa tgggcggctc ttcggccagc atggcttcta tgaccggtgg
     1501 ccaacagatg ggtggttcct cagctagaat ggcgagtatg accggtggcc aacagatggg
     1561 tggtagtgga agcgcgtgga gccaccctca gttcgagaag ggaggaggaa gcggcggagg
     1621 cagcggagga agcgcttgga gccacccgca gttcgagaaa ggagctagat cacccgggct
     1681 cgagtgagca gttaacgaat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg
     1741 agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg
     1801 cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc
     1861 gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg
     1921 caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc
     1981 tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc
     2041 tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag
     2101 gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg
     2161 cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct gcatcccgcc
     2221 accatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc
     2281 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc
     2341 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa
     2401 gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac
     2461 attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg
     2521 ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg
     2581 gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa
     2641 ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg
     2701 tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat
     2761 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc
     2821 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag
     2881 gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg
     2941 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg
     3001 ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt
     3061 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc
     3121 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat
     3181 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca
     3241 aaggaataga gtagatgccg accgaacaag agctgatttc gagaacgcct cagccagcaa
     3301 ctcgcgcgag cctagcaagg caaatgcgag agaacggcct tacgcttggt ggcacagttc
     3361 tcgtccacag ttcgctaagc tcgctcggct gggtcgcggg agggccggtc gcagtgattc
     3421 aggcccttct ggattgtgtt ggtccccagg gcacgattgt catgcccacg cactcgggtg
     3481 atctgactga tcccgcagat tggagatcgc cgcccgtgcc tgccgattgg gtgcagatcc
     3541 gtcgacctgc agccaagctt atcgataaaa taaaagattt tatttagtct ccagaaaaag
     3601 gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc
     3661 aaggcatgga aaatacataa ctgagaatag agaagttcag atcaaggtta ggaacagaga
     3721 gacagcagaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
     3781 gccaagaaca gatggtcccc agatgcggtc ccgccctcag cagtttctag agaaccatca
     3841 gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa
     3901 tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc
     3961 acaacccctc actcggcgcg ccagtcctcc gatagactgc gtcgcccggg tacccgtgta
     4021 tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc
     4081 tcctctgagt gattgactac ccgtcagcgg gggtctttca tgggtaacag tttcttgaag
     4141 ttggagaaca acattctgag ggtaggagtc gaatattaag taatcctgac tcaattagcc
     4201 actgttttga atccacatac tccaatactc ctgaaatagt tcattatgga cagcgcagaa
     4261 agagctgggg agaattgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4321 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4381 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4441 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4501 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4561 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4621 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4681 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4741 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4801 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4861 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4921 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4981 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5041 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5101 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5161 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5221 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5281 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5341 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5401 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5461 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5521 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5581 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5641 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5701 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5761 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5821 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5881 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5941 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6001 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6061 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6121 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6181 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6241 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6301 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6361 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     6421 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     6481 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     6541 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     6601 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     6661 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     6721 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     6781 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     6841 tttcccagtc acgacgttgt aaaacgacgg cgcaaggaat ggtgcatgca aggagatggc
     6901 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     6961 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     7021 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggcgattagt
     7081 ccaatttgtt aaagacagga tatcagtggt ccaggctcta gttttgactc aacaatatca
     7141 ccagctgaag cctatagagt acgagccata gataaaataa aagattttat ttagtctcca
     7201 gaaaaagggg gg
//
