LOCUS       sequence-438515-        7568 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of TASL-NEMO IKK binding site in mammalian cells by 
            retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7568)
  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 7568)
  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-TASL-NEMO
FEATURES             Location/Qualifiers
     source          1..7568
                     /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    1191..1757
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1207..1759
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1351..1368)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1578..1597
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1771..1773,1774..1776,1777..2490)
                     /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(1816..1837)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2077..2096)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2424..2445
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2552..3066
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3228..3244
                     /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(3252..3258,3259..3276,3277..3282))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3297..3318
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3435..3452)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3606..4194)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3686..3705)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4365..5156,5157..5225))
                     /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     4988..5007
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5226..5330)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5398..5416
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5454..5476)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5576..5595
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5789..5811
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5939..5958
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6158..6674
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6738..7079
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             7146..7562
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7492..7514
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7530..7546
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 agatctgcca ccatggatta caaagacgat gatgacaaga tgttgtctga agggtacttg
       61 tcaggattga cttattggaa cgacatccat tggaactgcg cttcctataa tgagcctgta
      121 gcaggtgatc aagggaagga gacctcatcc gtcgcagctt tgagctatag ttctgtcgac
      181 gagacccagg ttcaaagcct ctatgtctct tgtaaaagtt ctggcaagtt tatatcttct
      241 gtccacgccc gagcttctca acactctcgg tcccagagtc gcactgtcct ccaggccaac
      301 agcaatccag tcttcgaaag cccaacactg gcagcagtgg gtatctgcag agatgtgatc
      361 agggaaacct atctcgtacc tccaagttgt aaaagtattt gcaagaatta caatgatctt
      421 cacatagcag gagggcaggt aatggctatt aacagtgtga tggccaactt tcctagcgag
      481 tcttccttcg aagacggacc cctgctcaag agttccgaaa tctcactgtc aatggaggac
      541 agcacctcca cccagcttac agaacttcct ctcaagccaa tccagagata ttcatcctac
      601 tggcgaatta cctcaatcaa agaaaagtca tctcttcaaa tgcagaagcc aatcagtaac
      661 gcagttctga acgagtactt ggaacagaaa gtcgtagagc tgtacaagca atatatcatg
      721 gacactgttt tccatgactc ttcaccaact cagatactgg catctgagtt tatcatgaca
      781 aatgtggatc agatttctct ccaagttagc aaagaaaaaa atctggatac ttcaaaagtt
      841 aaagatatag tgatttctca tctcttgcaa ctcgtaagta gtgaaataag cacacccagc
      901 cttcatatct cacagtacag caatataact cccgaacagg ggacaccaga gacattgcaa
      961 cgatgtctgg aagaaaatca ggaacttaga gatgctattc gccaaagcaa ccaaatgctg
     1021 cgggagcgct gtgaagagct cctgcacttt caagtctcac aaagggaaga aaaagagttt
     1081 ctgatgtgta aattccagga agctcgaaag ttggtcgaac ggctctcact ggagaagtga
     1141 taagcggccg cgtttaaacg tcgacggtat cgataagctt gatatcgaat tccgcccccc
     1201 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
     1261 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
     1321 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
     1381 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
     1441 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
     1501 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
     1561 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
     1621 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg
     1681 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     1741 gaaaaacacg atgataatat ggccacaacc atggtgagca agggcgagga gctgttcacc
     1801 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1861 tctggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     1921 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     1981 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     2041 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     2101 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     2161 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     2221 gtctatatca tggccgacaa gcagaagaac ggcatcaagg cgaacttcaa gatccgccac
     2281 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     2341 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     2401 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     2461 actctcggca tggacgagct gtacaagtaa tgaattaatt aagaattcga taaaataaaa
     2521 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     2581 gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag
     2641 ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg
     2701 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc
     2761 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
     2821 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
     2881 ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag
     2941 actgcgtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt
     3001 ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc
     3061 tttcatgggt aacagtttct tgaagttgga gaacaacatt ctgagggtag gagtcgaata
     3121 ttaagtaatc ctgactcaat tagccactgt tttgaatcca catactccaa tactcctgaa
     3181 atagttcatt atggacagcg cagaaagagc tggggagaat tgtgaaattg ttatccgctc
     3241 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     3301 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3361 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3421 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3481 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3541 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3601 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3661 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3721 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3781 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3841 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3901 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3961 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4021 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4081 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4141 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4201 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4261 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4321 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4381 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4441 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4501 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4561 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4621 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4681 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4741 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4801 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4861 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4921 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4981 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5041 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5101 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5161 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5221 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5281 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5341 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     5401 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     5461 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     5521 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     5581 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     5641 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     5701 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     5761 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggcgcaa
     5821 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     5881 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     5941 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     6001 tgcgtccggc gtagaggcga ttagtccaat ttgttaaaga caggatatca gtggtccagg
     6061 ctctagtttt gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagataa
     6121 aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt
     6181 ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat
     6241 agagaagttc agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata
     6301 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg
     6361 tcccgccctc agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg
     6421 aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc
     6481 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct
     6541 ccgatagact gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc
     6601 cgaatcgtgg actcgctgat ccttgggagg gtctcctcag attgattgac tgcccacctc
     6661 gggggtcttt catttggagg ttccaccgag atttggagac ccctgcccag ggaccaccga
     6721 cccccccgcc gggaggtaag ctggccagcg gtcgtttcgt gtctgtctct gtctttgtgc
     6781 gtgtttgtgc cggcatctaa tgtttgcgcc tgcgtctgta ctagttagct aactagctct
     6841 gtatctggcg gacccgtggt ggaactgacg agttctgaac acccggccgc aaccctggga
     6901 gacgtcccag ggactttggg ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt
     6961 ggaatccgac cccgtcagga tatgtggttc tggtaggaga cgagaaccta aaacagttcc
     7021 cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc
     7081 agcgctgcag catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa
     7141 attagggcca gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg
     7201 agcggatcgc tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct
     7261 ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc
     7321 tcatcaccca ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg
     7381 tcccctacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct
     7441 ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac
     7501 ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg
     7561 ccggaatt
//
