LOCUS       sequence-439081-        7937 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of IRSp53 C-terminal truncation 4 (247-295) in mammalian 
            cells by retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7937)
  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 7937)
  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-IRSp53(trunc C4)
FEATURES             Location/Qualifiers
     source          1..7937
                     /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    1560..2126
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1576..2128
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1720..1737)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1947..1966
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2140..2142,2143..2145,2146..2859)
                     /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(2185..2206)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2446..2465)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2793..2814
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2921..3435
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3597..3613
                     /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(3621..3627,3628..3645,3646..3651))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3666..3687
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3804..3821)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3975..4563)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4055..4074)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4734..5525,5526..5594))
                     /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     5357..5376
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5595..5699)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5767..5785
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5823..5845)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5945..5964
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6158..6180
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6308..6327
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6527..7043
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7107..7448
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             7515..7931
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7861..7883
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7899..7915
                     /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 gctggggcta tggccaatag caatggctcc
     1381 atccttccaa gtgccctgtc agcttccaag tccaacctgg tcatctcaga tcccattcct
     1441 ggagccaagc ccttgccagt gccacctgag ctggcaccat ttgtggggcg gatgtctgct
     1501 caggagtgat aagcggccgc gtttaaacgt cgacggtatc gataagcttg atatcgaatt
     1561 ccgccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     1621 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     1681 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     1741 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     1801 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     1861 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     1921 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     1981 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct
     2041 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     2101 ttttcctttg aaaaacacga tgataatatg gccacaacca tggtgagcaa gggcgaggag
     2161 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     2221 ttcagcgtgt ctggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     2281 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     2341 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     2401 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     2461 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     2521 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     2581 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggc gaacttcaag
     2641 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     2701 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     2761 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2821 gccgggatca ctctcggcat ggacgagctg tacaagtaat gaattaatta agaattcgat
     2881 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
     2941 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac ataactgaga
     3001 atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg ccaaacagga
     3061 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc
     3121 ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc
     3181 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
     3241 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
     3301 ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
     3361 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
     3421 gcgggggtct ttcatgggta acagtttctt gaagttggag aacaacattc tgagggtagg
     3481 agtcgaatat taagtaatcc tgactcaatt agccactgtt ttgaatccac atactccaat
     3541 actcctgaaa tagttcatta tggacagcgc agaaagagct ggggagaatt gtgaaattgt
     3601 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3661 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3721 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3781 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3841 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3901 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3961 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4021 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4081 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4141 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4201 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4261 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4321 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4381 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4441 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4501 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4561 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4621 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4681 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4741 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4801 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4861 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4921 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4981 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5041 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5101 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5161 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5221 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5281 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5341 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5401 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5461 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5521 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5581 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5641 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5701 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     5761 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     5821 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     5881 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     5941 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     6001 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     6061 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     6121 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     6181 acggcgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
     6241 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
     6301 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
     6361 cggccacgat gcgtccggcg tagaggcgat tagtccaatt tgttaaagac aggatatcag
     6421 tggtccaggc tctagttttg actcaacaat atcaccagct gaagcctata gagtacgagc
     6481 catagataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     6541 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaatacata
     6601 actgagaata gagaagttca gatcaaggtt aggaacagag agacagcaga atatgggcca
     6661 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     6721 cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     6781 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     6841 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc
     6901 gccagtcctc cgatagactg cgtcgcccgg gtacccgtat tcccaataaa gcctcttgct
     6961 gtttgcatcc gaatcgtgga ctcgctgatc cttgggaggg tctcctcaga ttgattgact
     7021 gcccacctcg ggggtctttc atttggaggt tccaccgaga tttggagacc cctgcccagg
     7081 gaccaccgac ccccccgccg ggaggtaagc tggccagcgg tcgtttcgtg tctgtctctg
     7141 tctttgtgcg tgtttgtgcc ggcatctaat gtttgcgcct gcgtctgtac tagttagcta
     7201 actagctctg tatctggcgg acccgtggtg gaactgacga gttctgaaca cccggccgca
     7261 accctgggag acgtcccagg gactttgggg gccgtttttg tggcccgacc tgaggaaggg
     7321 agtcgatgtg gaatccgacc ccgtcaggat atgtggttct ggtaggagac gagaacctaa
     7381 aacagttccc gcctccgtct gaatttttgc tttcggtttg gaaccgaagc cgcgcgtctt
     7441 gtctgctgca gcgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt
     7501 tgtctgaaaa ttagggccag actgttacca ctcccttaag tttgacctta ggtcactgga
     7561 aagatgtcga gcggatcgct cacaaccagt cggtagatgt caagaagaga cgttgggtta
     7621 ccttctgctc tgcagaatgg ccaaccttta acgtcggatg gccgcgagac ggcaccttta
     7681 accgagacct catcacccag gttaagatca aggtcttttc acctggcccg catggacacc
     7741 cagaccaggt cccctacatc gtgacctggg aagccttggc ttttgacccc cctccctggg
     7801 tcaagccctt tgtacaccct aagcctccgc ctcctcttcc tccatccgcc ccgtctctcc
     7861 cccttgaacc tcctcgttcg accccgcctc gatcctccct ttatccagcc ctcactcctt
     7921 ctctaggcgc cggaatt
//
