LOCUS       sequence-439049-        8147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of IRSp53 N-terminal truncation 1 (257-375) in mammalian 
            cells by retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8147)
  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 8147)
  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 N1)
FEATURES             Location/Qualifiers
     source          1..8147
                     /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    1770..2336
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1786..2338
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1930..1947)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2157..2176
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2350..2352,2353..2355,2356..3069)
                     /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(2395..2416)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2656..2675)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3003..3024
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3131..3645
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3807..3823
                     /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(3831..3837,3838..3855,3856..3861))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3876..3897
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4014..4031)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4185..4773)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4265..4284)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4944..5735,5736..5804))
                     /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     5567..5586
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5805..5909)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5977..5995
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6033..6055)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6155..6174
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6368..6390
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6518..6537
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6737..7253
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7317..7658
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             7725..8141
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8071..8093
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8109..8125
                     /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 gtgccctgtc agcttccaag
     1381 tccaacctgg tcatctcaga tcccattcct ggagccaagc ccttgccagt gccacctgag
     1441 ctggcaccat ttgtggggcg gatgtctgct caggagaatg tacctgtcat gaatggcgtc
     1501 gcaggcccgg acagcgagga ctacaacccc tgggctgacc gaaaggctgc ccagcccaaa
     1561 tccctgtctc ccccgcagtc tcaaagcaag ctgagcgact cgtactccaa cacactcccc
     1621 gtgcgcaaga gcgtgacgcc gaagaacagc tatgccacca ctgagaacaa gacactgcca
     1681 cgttcaagct ccatggcagc tggcctagaa cgtaattgat aagcggccgc gtttaaacgt
     1741 cgacggtatc gataagcttg atatcgaatt ccgccccccc ccctaacgtt actggccgaa
     1801 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
     1861 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
     1921 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
     1981 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
     2041 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
     2101 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
     2161 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
     2221 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac
     2281 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg
     2341 gccacaacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     2401 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ctggcgaggg cgagggcgat
     2461 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     2521 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     2581 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     2641 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2701 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2761 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     2821 cagaagaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga cggcagcgtg
     2881 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     2941 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     3001 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     3061 tacaagtaat gaattaatta agaattcgat aaaataaaag attttattta gtctccagaa
     3121 aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt
     3181 ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca
     3241 gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     3301 cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc
     3361 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa
     3421 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga
     3481 gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg
     3541 tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag
     3601 ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatgggta acagtttctt
     3661 gaagttggag aacaacattc tgagggtagg agtcgaatat taagtaatcc tgactcaatt
     3721 agccactgtt ttgaatccac atactccaat actcctgaaa tagttcatta tggacagcgc
     3781 agaaagagct ggggagaatt gtgaaattgt tatccgctca caattccaca caacatacga
     3841 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3901 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3961 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4021 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4081 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4141 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4201 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4261 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4321 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4381 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4441 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4501 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4561 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4621 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4681 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4741 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4801 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4861 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4921 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4981 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5041 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5101 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5161 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5221 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5281 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5341 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5401 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5461 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5521 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5581 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5641 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5701 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5761 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5821 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5881 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     5941 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     6001 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     6061 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     6121 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     6181 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     6241 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     6301 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     6361 gggttttccc agtcacgacg ttgtaaaacg acggcgcaag gaatggtgca tgcaaggaga
     6421 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     6481 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     6541 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggcgat
     6601 tagtccaatt tgttaaagac aggatatcag tggtccaggc tctagttttg actcaacaat
     6661 atcaccagct gaagcctata gagtacgagc catagataaa ataaaagatt ttatttagtc
     6721 tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa
     6781 cgccattttg caaggcatgg aaaatacata actgagaata gagaagttca gatcaaggtt
     6841 aggaacagag agacagcaga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     6901 cccggctcag ggccaagaac agatggtccc cagatgcggt cccgccctca gcagtttcta
     6961 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
     7021 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
     7081 taaaagagcc cacaacccct cactcggcgc gccagtcctc cgatagactg cgtcgcccgg
     7141 gtacccgtat tcccaataaa gcctcttgct gtttgcatcc gaatcgtgga ctcgctgatc
     7201 cttgggaggg tctcctcaga ttgattgact gcccacctcg ggggtctttc atttggaggt
     7261 tccaccgaga tttggagacc cctgcccagg gaccaccgac ccccccgccg ggaggtaagc
     7321 tggccagcgg tcgtttcgtg tctgtctctg tctttgtgcg tgtttgtgcc ggcatctaat
     7381 gtttgcgcct gcgtctgtac tagttagcta actagctctg tatctggcgg acccgtggtg
     7441 gaactgacga gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg
     7501 gccgtttttg tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat
     7561 atgtggttct ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc
     7621 tttcggtttg gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt
     7681 gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca
     7741 ctcccttaag tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt
     7801 cggtagatgt caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta
     7861 acgtcggatg gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca
     7921 aggtcttttc acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg
     7981 aagccttggc ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc
     8041 ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc
     8101 gatcctccct ttatccagcc ctcactcctt ctctaggcgc cggaatt
//
