LOCUS       sequence-490853-        9080 bp    DNA     circular SYN 28-OCT-2025
DEFINITION  Expression of SYNDAPIN2 (pLxIS) in mammalian cells by retroviral 
            transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9080)
  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 9080)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 28, 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-SYNDAPIN2
FEATURES             Location/Qualifiers
     source          1..9080
                     /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    2703..3269
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2719..3271
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2863..2880)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3090..3109
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3283..3285,3286..3288,3289..4002)
                     /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(3328..3349)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3589..3608)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3936..3957
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4064..4578
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4740..4756
                     /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(4764..4770,4771..4788,4789..4794))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4809..4830
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4947..4964)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5118..5706)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5198..5217)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5877..6668,6669..6737))
                     /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     6500..6519
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6738..6842)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6910..6928
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6966..6988)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7088..7107
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7301..7323
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7451..7470
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             7670..8186
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    8250..8591
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             8658..9074
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     9004..9026
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     9042..9058
                     /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 tgtctgtgac ttatgatgac
     1381 agcgtcggtg tcgaggtgtc ttctgactct ttttgggaag tgggtaacta caagcgaacc
     1441 gtaaagagga tagacgacgg ccacagattg tgcggtgatc tgatgaattg cctccatgag
     1501 cgcgctcgaa ttgagaaggc ctacgcacag caacttactg agtgggcaag acgatggcga
     1561 caactggtcg aaaaggggcc ccaatacgga acagtagaaa aggcttggat cgcagttatg
     1621 tcagaggctg aacgcgtctc agagcttcat ttggaagtca aggcttctct tatgaatgaa
     1681 gattttgaaa aaataaaaaa ttggcaaaaa gaagccttcc acaagcaaat gatgggcggc
     1741 tttaaggaaa ctaaagaagc tgaagatggc tttagaaaag cacagaagcc ctgggccaaa
     1801 aagttgaagg aagtggaggc tgccaagaaa gctcaccata ctgcatgtaa agaagaaaag
     1861 ctcgctattt caagggaggc taactcaaaa gccgatccct cccttaaccc cgaacaattg
     1921 aagaagctcc aggacaagat cgagaagtgc aagcaagacg tgctcaaaac caaagataaa
     1981 tatgagaagt ctttgaaaga attggatcaa accacccctc agtacatgga aaacatggag
     2041 caggtgttcg aacagtgcca acagtttgaa gagaaaagac ttcgcttttt ccgagaggtt
     2101 ctcttggagg tgcagaagca cctggatctc agcaatgtgg cttcctataa aacaatctac
     2161 agagaattgg aacaatccat aaaggctgcc gatgcagtgg aagatttgcg atggtttaga
     2221 gcaaatcacg ggccaggaat ggcaatgaac tggccacaat ttgaagaatg gtctgctgat
     2281 ctgaaccgga ctttgagccg ccgggagaaa aaaaaagcag tggacggagt gacccttaca
     2341 gggatcaatc aaaccggcga ccagagcggc caaaacaaac caggttcaaa cttgtcagta
     2401 cccagcaacc cagcccaaag tactcaactg caatcatctt acaacccttt cgaagatgaa
     2461 gatgatacag gaagctccat aagcgagaaa gaggatatca aggcaaaaaa tgtgtcctct
     2521 tacgagaaga ctcaaacata tccaactgat tggtccgacg acgaatcaaa taatcccttc
     2581 agtagtacag acgccaacgg tgactccaat ccttttgatg aggacacaac cagtggtaca
     2641 gaggtgaggt gataagcggc cgcgtttaaa cgtcgacggt atcgataagc ttgatatcga
     2701 attccgcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     2761 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     2821 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     2881 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     2941 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     3001 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     3061 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     3121 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     3181 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     3241 tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggtgag caagggcgag
     3301 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     3361 aagttcagcg tgtctggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     3421 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     3481 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     3541 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     3601 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     3661 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     3721 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggcgaacttc
     3781 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     3841 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     3901 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     3961 gccgccggga tcactctcgg catggacgag ctgtacaagt aatgaattaa ttaagaattc
     4021 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg
     4081 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa tacataactg
     4141 agaatagaga agttcagatc aaggttagga acagagagac agcagaatat gggccaaaca
     4201 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
     4261 tgcggtcccg ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
     4321 acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt
     4381 cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact cggcgcgcca
     4441 gtcctccgat agactgcgtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg
     4501 catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg
     4561 tcagcggggg tctttcatgg gtaacagttt cttgaagttg gagaacaaca ttctgagggt
     4621 aggagtcgaa tattaagtaa tcctgactca attagccact gttttgaatc cacatactcc
     4681 aatactcctg aaatagttca ttatggacag cgcagaaaga gctggggaga attgtgaaat
     4741 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4801 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4861 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4921 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4981 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5041 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5101 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5161 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5221 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5281 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5341 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5401 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5461 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5521 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5581 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5641 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5701 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5761 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5821 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5881 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5941 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6001 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6061 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6121 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6181 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6241 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6301 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6361 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6421 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6481 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6541 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6601 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6661 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6721 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6781 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6841 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     6901 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     6961 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     7021 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     7081 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     7141 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
     7201 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     7261 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     7321 acgacggcgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     7381 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     7441 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     7501 tgccggccac gatgcgtccg gcgtagaggc gattagtcca atttgttaaa gacaggatat
     7561 cagtggtcca ggctctagtt ttgactcaac aatatcacca gctgaagcct atagagtacg
     7621 agccatagat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
     7681 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac
     7741 ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg
     7801 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
     7861 ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
     7921 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     7981 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     8041 cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat aaagcctctt
     8101 gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc agattgattg
     8161 actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag acccctgccc
     8221 agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc gtgtctgtct
     8281 ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg tactagttag
     8341 ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga acacccggcc
     8401 gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg acctgaggaa
     8461 gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga gacgagaacc
     8521 taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga agccgcgcgt
     8581 cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt
     8641 atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc ttaggtcact
     8701 ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag agacgttggg
     8761 ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga gacggcacct
     8821 ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc ccgcatggac
     8881 acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac ccccctccct
     8941 gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc gccccgtctc
     9001 tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca gccctcactc
     9061 cttctctagg cgccggaatt
//
