LOCUS       sequence-439091-        8393 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ARHGAP45 (pLxIS) in mammalian cells by retroviral 
            transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8393)
  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 8393)
  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-ARHGAP45
FEATURES             Location/Qualifiers
     source          1..8393
                     /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    2016..2582
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2032..2584
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2176..2193)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2403..2422
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2596..2598,2599..2601,2602..3315)
                     /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(2641..2662)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2902..2921)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3249..3270
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3377..3891
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4053..4069
                     /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(4077..4083,4084..4101,4102..4107))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4122..4143
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4260..4277)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4431..5019)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4511..4530)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5190..5981,5982..6050))
                     /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     5813..5832
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6051..6155)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6223..6241
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6279..6301)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6401..6420
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6614..6636
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6764..6783
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6983..7499
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7563..7904
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             7971..8387
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8317..8339
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8355..8371
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 agatctgcca ccatggacta caaagacgat gacgacaagg gcgtccaagt ggagacaatc
       61 tctcctgggg atggaaggac attccccaag cggggtcaga cctgcgttgt tcattatacc
      121 ggtatgttgg aagacggtaa aaaagtcgat agctcacggg atcgcaataa gcctttcaag
      181 ttcatgcttg ggaaacagga agtaatacgc ggctgggaag agggagtagc tcaaatgtcc
      241 gtcggtcaac gcgccaaatt gacaatttct cctgactacg cttatggtgc aaccggacac
      301 cccggcataa ttcccccaca cgcaacactt gtatttgatg tagaattgct caagctggaa
      361 acaagaggag tccaggttga aaccatatct cctggggacg gaagaacctt ccctaagcgg
      421 ggacaaactt gcgttgtcca ttacacaggc atgcttgaag atggtaagaa agtcgactca
      481 tcacgagaca gaaataagcc attcaaattt atgctcggta aacaggaagt cataagaggc
      541 tgggaagaag gagtggccca aatgtctgta ggacaacgag ccaaactcac aatatctccc
      601 gattatgctt atggtgccac tggccacccc ggcattatac cacctcatgc cactcttgtt
      661 tttgacgtag agctgttgaa gttggagacc cggggcgtgc aggttgagac tataagtcct
      721 ggagatggaa ggacttttcc taaaagaggt cagacatgcg tagtacacta tacagggatg
      781 ttggaggacg gcaaaaaagt cgacagctcc cgagaccgga ataagccttt taagttcatg
      841 ttggggaaac aggaagtaat ccgcgggtgg gaagagggag tcgctcagat gtctgtggga
      901 cagcgcgcta agctgaccat ttcccccgat tatgcctacg gcgctaccgg tcaccccggg
      961 attattccac ctcacgctac tttggtgttc gatgtcgaac ttctgaaact ggagactcgg
     1021 ggagtacagg ttgaaacaat cagcccagga gacggaagaa cctttccaaa aaggggtcaa
     1081 acatgtgtgg tacactacac aggaatgttg gaagacggaa aaaaggtgga cagctctcga
     1141 gaccgcaata aacctttcaa gtttatgctg ggcaaacagg aggtgatacg cggatgggag
     1201 gaaggcgtag cccagatgtc cgtgggtcag cgggcaaaac ttacaatttc acccgattac
     1261 gcatatggcg caacaggaca tccaggtatt ataccaccac atgccaccct tgttttcgac
     1321 gtagagcttt tgaaactcga aggggctgga gcaggcgcta gcaaattgta tgaccccggc
     1381 caacaatacg catcccatgt gcgccaactg caacggggag aagaaccaga tgtccgatat
     1441 gacttcgagc cttacgtgag caataacagc tggagtccca taatgaggac acggaagggc
     1501 tcattcaatc caggggatgc aagcggaccc gaggcagctg gtagtcctcc cgaagaaggc
     1561 gggacatctg aagcagcacc aaacaaagac catagaggcg gtcgcggcca tcaagttcat
     1621 aagtcatggc ccatatctat cagtgacacc gaagttggtc ttgatacatc aagcggtgat
     1681 ttgaagaaat tcgaccgaac aagtagcagt gggaccatga gcagttctga ggaattggga
     1741 gatcaggagg ctggtcttgt cgcctccgcc ttcgatagtg cagacctcaa cggcatggac
     1801 ccagagttgc ccgttgctat gccatccggt ccctttcggc acgtaggcct tagcaaggcc
     1861 gctaggacac atcggctgag gaaacttagg accccagcta aatgtaggga atgtaatagc
     1921 tatgtttact tccaaggagc agagtgtgaa gagtgctgtc tctgataagc ggccgcgttt
     1981 aaacgtcgac ggtatcgata agcttgatat cgaattccgc ccccccccct aacgttactg
     2041 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat
     2101 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc
     2161 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag
     2221 cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc
     2281 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac
     2341 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca
     2401 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt
     2461 gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa
     2521 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat
     2581 aatatggcca caaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     2641 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtctgg cgagggcgag
     2701 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     2761 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     2821 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     2881 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     2941 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     3001 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     3061 gacaagcaga agaacggcat caaggcgaac ttcaagatcc gccacaacat cgaggacggc
     3121 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     3181 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     3241 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     3301 gagctgtaca agtaatgaat taattaagaa ttcgataaaa taaaagattt tatttagtct
     3361 ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac
     3421 gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag atcaaggtta
     3481 ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     3541 ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag cagtttctag
     3601 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
     3661 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
     3721 aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc gtcgcccggg
     3781 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
     3841 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tgggtaacag
     3901 tttcttgaag ttggagaaca acattctgag ggtaggagtc gaatattaag taatcctgac
     3961 tcaattagcc actgttttga atccacatac tccaatactc ctgaaatagt tcattatgga
     4021 cagcgcagaa agagctgggg agaattgtga aattgttatc cgctcacaat tccacacaac
     4081 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4141 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4201 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4261 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4321 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4381 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4441 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4501 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4561 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4621 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4681 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4741 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4801 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4861 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4921 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4981 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5041 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5101 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5161 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5221 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5281 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5341 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5401 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5461 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5521 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5581 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5641 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5701 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5761 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5821 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5881 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5941 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6001 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6061 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6121 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6181 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     6241 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     6301 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     6361 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     6421 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     6481 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     6541 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     6601 acgccagggt tttcccagtc acgacgttgt aaaacgacgg cgcaaggaat ggtgcatgca
     6661 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     6721 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
     6781 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
     6841 ggcgattagt ccaatttgtt aaagacagga tatcagtggt ccaggctcta gttttgactc
     6901 aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa aagattttat
     6961 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt
     7021 aagtaacgcc attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc
     7081 aaggttagga acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt
     7141 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag
     7201 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
     7261 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
     7321 gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc
     7381 gcccgggtac ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggactcg
     7441 ctgatccttg ggagggtctc ctcagattga ttgactgccc acctcggggg tctttcattt
     7501 ggaggttcca ccgagatttg gagacccctg cccagggacc accgaccccc ccgccgggag
     7561 gtaagctggc cagcggtcgt ttcgtgtctg tctctgtctt tgtgcgtgtt tgtgccggca
     7621 tctaatgttt gcgcctgcgt ctgtactagt tagctaacta gctctgtatc tggcggaccc
     7681 gtggtggaac tgacgagttc tgaacacccg gccgcaaccc tgggagacgt cccagggact
     7741 ttgggggccg tttttgtggc ccgacctgag gaagggagtc gatgtggaat ccgaccccgt
     7801 caggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat
     7861 ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg
     7921 ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaattag ggccagactg
     7981 ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca
     8041 accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa
     8101 cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta
     8161 agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga
     8221 cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc
     8281 ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc
     8341 cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccgga att
//
