LOCUS       sequence-439108-        8363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ASAP2 (pLxIS) in mammalian cells by retroviral 
            transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8363)
  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 8363)
  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-ASAP2
FEATURES             Location/Qualifiers
     source          1..8363
                     /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    1986..2552
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2002..2554
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2146..2163)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2373..2392
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2566..2568,2569..2571,2572..3285)
                     /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(2611..2632)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2872..2891)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3219..3240
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3347..3861
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4023..4039
                     /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(4047..4053,4054..4071,4072..4077))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4092..4113
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4230..4247)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4401..4989)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4481..4500)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5160..5951,5952..6020))
                     /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     5783..5802
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6021..6125)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6193..6211
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6249..6271)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6371..6390
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6584..6606
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6734..6753
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6953..7469
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7533..7874
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             7941..8357
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8287..8309
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8325..8341
                     /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 ccgacttgca caccataaaa
     1381 caagcccaag acgaggagcg gaggcaactt atacagctta gggatatact caaaagcgcc
     1441 ctgcaagtcg aacagaaaga gagcaggcgc gacagccagc ttcgccaaag cacagcctac
     1501 tctttgcatc aaccccaagg aaataaggag cacggaaccg aacggaacgg gaatctgtat
     1561 aaaaagtcag atgggatacg aaaggtttgg caaaaacgga agtgtagtgt aaaaaacgga
     1621 ttcctcacta taagtcatgg cactgccaac aggcctcccg ctaagctgaa tcttctcact
     1681 tgccaggtca agactaaccc agaggagaaa aaatgcttcg atctcattag tcatgaccgc
     1741 acataccatt tccaagctga ggatgaacaa gaatgtcaga tttggatgag cgttctgcag
     1801 aatagcaaag aggaagcctt gaataacgct ttcaaagggg atgacaatac aggagagaac
     1861 aatatagtgc aagaattgac taaagaaatt attagcgaag tacaacgcat gaccggaaat
     1921 gatgtatgtt gctgataagc ggccgcgttt aaacgtcgac ggtatcgata agcttgatat
     1981 cgaattccgc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     2041 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     2101 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     2161 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
     2221 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     2281 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
     2341 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
     2401 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca
     2461 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
     2521 acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatggt gagcaagggc
     2581 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2641 cacaagttca gcgtgtctgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     2701 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     2761 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     2821 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     2881 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2941 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     3001 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggcgaac
     3061 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     3121 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     3181 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     3241 accgccgccg ggatcactct cggcatggac gagctgtaca agtaatgaat taattaagaa
     3301 ttcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
     3361 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaatacataa
     3421 ctgagaatag agaagttcag atcaaggtta ggaacagaga gacagcagaa tatgggccaa
     3481 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc
     3541 agatgcggtc ccgccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca
     3601 aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct
     3661 gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg
     3721 ccagtcctcc gatagactgc gtcgcccggg tacccgtgta tccaataaac cctcttgcag
     3781 ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac
     3841 ccgtcagcgg gggtctttca tgggtaacag tttcttgaag ttggagaaca acattctgag
     3901 ggtaggagtc gaatattaag taatcctgac tcaattagcc actgttttga atccacatac
     3961 tccaatactc ctgaaatagt tcattatgga cagcgcagaa agagctgggg agaattgtga
     4021 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4081 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4141 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4201 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4261 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4321 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4381 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4441 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4501 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4561 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4621 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4681 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4741 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4801 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4861 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4921 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4981 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5041 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5101 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5161 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5221 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5281 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5341 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5401 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5461 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5521 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5581 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5641 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5701 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5761 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5821 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5881 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5941 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6001 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6061 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6121 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     6181 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     6241 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     6301 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     6361 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     6421 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
     6481 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     6541 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     6601 aaaacgacgg cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     6661 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     6721 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     6781 tgatgccggc cacgatgcgt ccggcgtaga ggcgattagt ccaatttgtt aaagacagga
     6841 tatcagtggt ccaggctcta gttttgactc aacaatatca ccagctgaag cctatagagt
     6901 acgagccata gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag
     6961 accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa
     7021 tacataactg agaatagaga agttcagatc aaggttagga acagagagac agcagaatat
     7081 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     7141 ggtccccaga tgcggtcccg ccctcagcag tttctagaga accatcagat gtttccaggg
     7201 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct
     7261 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact
     7321 cggcgcgcca gtcctccgat agactgcgtc gcccgggtac ccgtattccc aataaagcct
     7381 cttgctgttt gcatccgaat cgtggactcg ctgatccttg ggagggtctc ctcagattga
     7441 ttgactgccc acctcggggg tctttcattt ggaggttcca ccgagatttg gagacccctg
     7501 cccagggacc accgaccccc ccgccgggag gtaagctggc cagcggtcgt ttcgtgtctg
     7561 tctctgtctt tgtgcgtgtt tgtgccggca tctaatgttt gcgcctgcgt ctgtactagt
     7621 tagctaacta gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg
     7681 gccgcaaccc tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag
     7741 gaagggagtc gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga
     7801 acctaaaaca gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg
     7861 cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc
     7921 tgtatttgtc tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc
     7981 actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt
     8041 gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca
     8101 cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg
     8161 gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc
     8221 cctgggtcaa gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt
     8281 ctctccccct tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca
     8341 ctccttctct aggcgccgga att
//
