LOCUS       sequence-438978-        7790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of FLAG-4xFKBP scaffold in mammalian cells by retroviral 
            transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7790)
  AUTHORS   Landau LM, Chaudhary N, Tien YC, Rogozinska M, Joshi S, Yao C, 
            Crowley J, Hullahalli K, Campbell IW, Waldor MK, Haigis M, Kagan JC
  TITLE     pLxIS-containing domains are biochemically flexible regulators of 
            interferons and metabolism.
  JOURNAL   Mol Cell. 2024 Jul 11;84(13):2436-2454.e10. doi: 
            10.1016/j.molcel.2024.05.030. Epub 2024 Jun 25.
  PUBMED    38925114
REFERENCE   2  (bases 1 to 7790)
  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: 
            "10.1016/j.molcel.2024.05.030"; journalName: "Mol Cell"; date: 
            "2024-07-11- 11"; volume: "84"; issue: "13"; pages: "2436-2454.e10"
COMMENT     Sequence Label: pMSCV-FKBP-empty
FEATURES             Location/Qualifiers
     source          1..7790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      7..16
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             16..39
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    1413..1979
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1429..1981
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1573..1590)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1800..1819
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1993..1995,1996..1998,1999..2712)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2038..2059)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2299..2318)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2646..2667
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2774..3288
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3450..3466
                     /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(3474..3480,3481..3498,3499..3504))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3519..3540
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3657..3674)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3828..4416)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3908..3927)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4587..5378,5379..5447))
                     /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     5210..5229
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5448..5552)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5620..5638
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5676..5698)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5798..5817
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6011..6033
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6161..6180
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6380..6896
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6960..7301
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             7368..7784
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7714..7736
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7752..7768
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 agatctgcca ccatggacta caaggacgac gacgacaagg gcgttcaagt agaaacaata
       61 tctccaggag acgggcgtac atttcctaag cgtggccaaa cttgcgtagt ccattatacc
      121 ggcatgctgg aggacggtaa gaaggtcgat agctctagag accgtaacaa acctttcaaa
      181 tttatgctgg ggaaacagga ggttatccgt ggttgggagg aaggcgtggc ccaaatgtcc
      241 gttgggcagc gtgctaagtt aaccatctcg ccagattacg catatggggc tacaggccat
      301 cctgggatca ttcccccgca tgcgacactt gtgtttgatg tagagctgtt gaagttggag
      361 acgcggggag ttcaagtcga aacgatttca ccaggtgatg gacgcacctt tcctaaacgg
      421 ggacaaacgt gcgtcgtgca ttacaccggt atgctggaag acggcaagaa agtcgattca
      481 agccgcgata gaaataaacc cttcaaattc atgcttggaa aacaggaagt aatacgcgga
      541 tgggaagagg gtgtggcgca aatgtctgtc gggcaaagag ctaaactgac aataagtcct
      601 gattacgcgt acggtgcgac tgggcaccct ggtatcattc cgccgcacgc gacacttgtc
      661 ttcgacgtcg agttacttaa gcttgagacg cggggggtgc aggtagagac tatttcgccc
      721 ggtgacggaa gaactttccc aaagcgggga cagacctgtg ttgtccacta cactgggatg
      781 ttggaggatg gtaagaaagt tgactcaagt cgtgatcgca acaaaccctt taagtttatg
      841 cttgggaagc aggaagtcat cagaggttgg gaagaaggcg tagctcagat gtcagtggga
      901 caacgggcca aattaaccat cagtccagac tacgcctacg gagccaccgg tcacccaggg
      961 atcataccac ctcacgcaac cttagtgttc gatgtggaac ttttaaagtt agagacgcgc
     1021 ggtgtacaag tggaaacaat cagtccaggg gatgggagaa cattccccaa aagaggccag
     1081 acgtgtgtcg tgcactatac ggggatgtta gaagatggga aaaaggtcga ttcgtcacgc
     1141 gaccgcaaca aaccgtttaa attcatgttg ggaaagcagg aggtgattcg cggctgggag
     1201 gaaggtgtgg cacagatgag cgtgggccag agagcaaaac tgactatctc tcctgattat
     1261 gcctacggcg cgacgggcca ccctggcata attccacccc acgctaccct ggtattcgat
     1321 gtcgagcttc tgaagcttga gggtgctggg gctggggctt gataagcggc cgcgtttaaa
     1381 cgtcgacggt atcgataagc ttgatatcga attccgcccc cccccctaac gttactggcc
     1441 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     1501 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     1561 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag
     1621 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     1681 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
     1741 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     1801 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
     1861 tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa
     1921 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat
     1981 atggccacaa ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2041 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtctggcga gggcgagggc
     2101 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     2161 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     2221 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     2281 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     2341 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     2401 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     2461 aagcagaaga acggcatcaa ggcgaacttc aagatccgcc acaacatcga ggacggcagc
     2521 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     2581 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     2641 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     2701 ctgtacaagt aatgaattaa ttaagaattc gataaaataa aagattttat ttagtctcca
     2761 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     2821 attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga
     2881 acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     2941 gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga
     3001 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     3061 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     3121 agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac
     3181 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     3241 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcatgg gtaacagttt
     3301 cttgaagttg gagaacaaca ttctgagggt aggagtcgaa tattaagtaa tcctgactca
     3361 attagccact gttttgaatc cacatactcc aatactcctg aaatagttca ttatggacag
     3421 cgcagaaaga gctggggaga attgtgaaat tgttatccgc tcacaattcc acacaacata
     3481 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3541 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3601 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3661 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3721 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3781 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3841 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3901 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3961 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4021 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4081 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4141 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4201 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4261 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4321 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4381 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4441 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4501 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4561 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4621 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4681 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4741 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4801 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4861 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4921 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4981 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5041 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5101 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5161 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5221 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5281 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5341 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5401 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5461 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5521 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5581 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     5641 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     5701 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     5761 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     5821 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     5881 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     5941 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     6001 ccagggtttt cccagtcacg acgttgtaaa acgacggcgc aaggaatggt gcatgcaagg
     6061 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     6121 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     6181 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagaggc
     6241 gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt ttgactcaac
     6301 aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag attttattta
     6361 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag
     6421 taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag
     6481 gttaggaaca gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc
     6541 tgccccggct cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt
     6601 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
     6661 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
     6721 caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc
     6781 cgggtacccg tattcccaat aaagcctctt gctgtttgca tccgaatcgt ggactcgctg
     6841 atccttggga gggtctcctc agattgattg actgcccacc tcgggggtct ttcatttgga
     6901 ggttccaccg agatttggag acccctgccc agggaccacc gacccccccg ccgggaggta
     6961 agctggccag cggtcgtttc gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct
     7021 aatgtttgcg cctgcgtctg tactagttag ctaactagct ctgtatctgg cggacccgtg
     7081 gtggaactga cgagttctga acacccggcc gcaaccctgg gagacgtccc agggactttg
     7141 ggggccgttt ttgtggcccg acctgaggaa gggagtcgat gtggaatccg accccgtcag
     7201 gatatgtggt tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt
     7261 tgctttcggt ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc
     7321 tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta
     7381 ccactccctt aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc
     7441 agtcggtaga tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct
     7501 ttaacgtcgg atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga
     7561 tcaaggtctt ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct
     7621 gggaagcctt ggcttttgac ccccctccct gggtcaagcc ctttgtacac cctaagcctc
     7681 cgcctcctct tcctccatcc gccccgtctc tcccccttga acctcctcgt tcgaccccgc
     7741 ctcgatcctc cctttatcca gccctcactc cttctctagg cgccggaatt
//
