LOCUS       sequence-439109-        8552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ASAP3 (Cterm pLxIS) in mammalian cells by retroviral 
            transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8552)
  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 8552)
  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-ASAP3(Cterm)
FEATURES             Location/Qualifiers
     source          1..8552
                     /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    2175..2741
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2191..2743
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2335..2352)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2562..2581
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2755..2757,2758..2760,2761..3474)
                     /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(2800..2821)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3061..3080)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3408..3429
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3536..4050
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4212..4228
                     /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(4236..4242,4243..4260,4261..4266))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4281..4302
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4419..4436)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4590..5178)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4670..4689)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5349..6140,6141..6209))
                     /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     5972..5991
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6210..6314)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6382..6400
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6438..6460)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6560..6579
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6773..6795
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6923..6942
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             7142..7658
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7722..8063
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             8130..8546
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8476..8498
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8514..8530
                     /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 atgatgccgg cgaaacagcc
     1381 ctggacattg cccgaaaccg ccaacataaa gagtgtgaag aacttcttga acaggcccag
     1441 gccggtactc tggctttccc acttcacatg gattaccact ggggtcatag catggagcac
     1501 ggtttcgatt cagaagagga ggaagaagag gagaagcatt gcccttccaa gccccccgct
     1561 caagcatgct ggggatccgt tcggctggac ataagtaaca aaacatacga aactgttgcc
     1621 actccaggcc ccgcaacaac acagtcccaa tctgaagact ctccccctcc attgcccatc
     1681 aaaaatagtt ctaggacaat cgtacttggc cgggccgggc actgtagtgg tgataggagt
     1741 gatttgcctt ctttgagatc cgaatcccct gaggccctcg agaatagatc cagtcctgct
     1801 tctagttcct ccagcctgac ctccagtgta gagcccggag gtttgagcca agccccaagc
     1861 tcccccgaag aaggtctgca agagagtgct tctatctccc gacctggcct cgcctctgga
     1921 actacctcag ctgaggttta tttgccagtc aagttcagta gcgagtcaac ccgctcctat
     1981 aggcgcgggg gcaggagcct tgaggactct ccatctgcac ggcagccttt gtgctctagg
     2041 aggcatattc ctgttggact cgtcgaagga gatgggagta aaatcggcgt cttgcctgat
     2101 agtttgcagc tgttgcatga ctgataagcg gccgcgttta aacgtcgacg gtatcgataa
     2161 gcttgatatc gaattccgcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     2221 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     2281 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     2341 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     2401 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     2461 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     2521 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     2581 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     2641 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     2701 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtg
     2761 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2821 gtaaacggcc acaagttcag cgtgtctggc gagggcgagg gcgatgccac ctacggcaag
     2881 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2941 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     3001 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     3061 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     3121 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     3181 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     3241 aaggcgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     3301 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     3361 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     3421 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaatgaatt
     3481 aattaagaat tcgataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa
     3541 agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa
     3601 aatacataac tgagaataga gaagttcaga tcaaggttag gaacagagag acagcagaat
     3661 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     3721 atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag atgtttccag
     3781 ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt
     3841 ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca
     3901 ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtgtat ccaataaacc
     3961 ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg
     4021 attgactacc cgtcagcggg ggtctttcat gggtaacagt ttcttgaagt tggagaacaa
     4081 cattctgagg gtaggagtcg aatattaagt aatcctgact caattagcca ctgttttgaa
     4141 tccacatact ccaatactcc tgaaatagtt cattatggac agcgcagaaa gagctgggga
     4201 gaattgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4261 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4321 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4381 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4441 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4501 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4561 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4621 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4681 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4741 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4801 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4861 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4921 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4981 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     5041 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5101 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5161 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5221 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5281 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5341 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5401 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5461 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5521 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5581 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5641 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5701 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5761 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5821 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5881 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5941 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6001 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6061 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6121 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6181 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6241 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6301 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     6361 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     6421 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     6481 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     6541 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     6601 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     6661 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     6721 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     6781 cgacgttgta aaacgacggc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     6841 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     6901 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     6961 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gcgattagtc caatttgtta
     7021 aagacaggat atcagtggtc caggctctag ttttgactca acaatatcac cagctgaagc
     7081 ctatagagta cgagccatag ataaaataaa agattttatt tagtctccag aaaaaggggg
     7141 gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg
     7201 catggaaaat acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca
     7261 gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     7321 agaacagatg gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg
     7381 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     7441 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     7501 cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtattccca
     7561 ataaagcctc ttgctgtttg catccgaatc gtggactcgc tgatccttgg gagggtctcc
     7621 tcagattgat tgactgccca cctcgggggt ctttcatttg gaggttccac cgagatttgg
     7681 agacccctgc ccagggacca ccgacccccc cgccgggagg taagctggcc agcggtcgtt
     7741 tcgtgtctgt ctctgtcttt gtgcgtgttt gtgccggcat ctaatgtttg cgcctgcgtc
     7801 tgtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttct
     7861 gaacacccgg ccgcaaccct gggagacgtc ccagggactt tgggggccgt ttttgtggcc
     7921 cgacctgagg aagggagtcg atgtggaatc cgaccccgtc aggatatgtg gttctggtag
     7981 gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc
     8041 gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt ctctgtctga
     8101 ctgtgtttct gtatttgtct gaaaattagg gccagactgt taccactccc ttaagtttga
     8161 ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga
     8221 agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc ggatggccgc
     8281 gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc ttttcacctg
     8341 gcccgcatgg acacccagac caggtcccct acatcgtgac ctgggaagcc ttggcttttg
     8401 acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct cttcctccat
     8461 ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc tccctttatc
     8521 cagccctcac tccttctcta ggcgccggaa tt
//
