LOCUS       sequence-438530-        8171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GARNL3 (pLxIS) in mammalian cells by retroviral 
            transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8171)
  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 8171)
  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-GARNL3
FEATURES             Location/Qualifiers
     source          1..8171
                     /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..81
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    1794..2360
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1810..2362
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1954..1971)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2181..2200
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2374..2376,2377..2379,2380..3093)
                     /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(2419..2440)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2680..2699)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3027..3048
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3155..3669
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3831..3847
                     /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(3855..3861,3862..3879,3880..3885))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3900..3921
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4038..4055)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4209..4797)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4289..4308)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4968..5759,5760..5828))
                     /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     5591..5610
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5829..5933)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6001..6019
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6057..6079)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6179..6198
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6392..6414
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6542..6561
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6761..7277
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7341..7682
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             7749..8165
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8095..8117
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8133..8149
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 agatctgcca ccatggacta caaggatcac gacggtgatt acaaggacca cgacatcgac
       61 tacaaggacg acgacgacaa gggcgttcaa gtagaaacaa tatctccagg agacgggcgt
      121 acatttccta agcgtggcca aacttgcgta gtccattata ccggcatgct ggaggacggt
      181 aagaaggtcg atagctctag agaccgtaac aaacctttca aatttatgct ggggaaacag
      241 gaggttatcc gtggttggga ggaaggcgtg gcccaaatgt ccgttgggca gcgtgctaag
      301 ttaaccatct cgccagatta cgcatatggg gctacaggcc atcctgggat cattcccccg
      361 catgcgacac ttgtgtttga tgtagagctg ttgaagttgg agacgcgggg agttcaagtc
      421 gaaacgattt caccaggtga tggacgcacc tttcctaaac ggggacaaac gtgcgtcgtg
      481 cattacaccg gtatgctgga agacggcaag aaagtcgatt caagccgcga tagaaataaa
      541 cccttcaaat tcatgcttgg aaaacaggaa gtaatacgcg gatgggaaga gggtgtggcg
      601 caaatgtctg tcgggcaaag agctaaactg acaataagtc ctgattacgc gtacggtgcg
      661 actgggcacc ctggtatcat tccgccgcac gcgacacttg tcttcgacgt cgagttactt
      721 aagcttgaga cgcggggggt gcaggtagag actatttcgc ccggtgacgg aagaactttc
      781 ccaaagcggg gacagacctg tgttgtccac tacactggga tgttggagga tggtaagaaa
      841 gttgactcaa gtcgtgatcg caacaaaccc tttaagttta tgcttgggaa gcaggaagtc
      901 atcagaggtt gggaagaagg cgtagctcag atgtcagtgg gacaacgggc caaattaacc
      961 atcagtccag actacgccta cggagccacc ggtcacccag ggatcatacc acctcacgca
     1021 accttagtgt tcgatgtgga acttttaaag ttagagacgc gcggtgtaca agtggaaaca
     1081 atcagtccag gggatgggag aacattcccc aaaagaggcc agacgtgtgt cgtgcactat
     1141 acggggatgt tagaagatgg gaaaaaggtc gattcgtcac gcgaccgcaa caaaccgttt
     1201 aaattcatgt tgggaaagca ggaggtgatt cgcggctggg aggaaggtgt ggcacagatg
     1261 agcgtgggcc agagagcaaa actgactatc tctcctgatt atgcctacgg cgcgacgggc
     1321 caccctggca taattccacc ccacgctacc ctggtattcg atgtcgagct tctgaagctt
     1381 gagggtgctg gggctggggc tatggatcca ctgaccaagg ggtcctgtgg gagtcagctg
     1441 gcacaaaccc tcctctggaa agcgaagtca tctctctctt tcggcatcca gccccttcag
     1501 acttggccaa caaaagatcc tgaactagag tcccaggtca acctctctgt ctcagaagac
     1561 ctaggctgta gacgtgggga tttcagtagg aagcattatg gatctgtgga gctgctcatc
     1621 tctagtgatg ccgatggagc catccaaagg gcaggaagat tccgagtgga gaatggctct
     1681 acagatgaga gtgcagctgc tctgccaggc acttggcgaa ggacagacgt gcacttagaa
     1741 tgataagcgg ccgcgtttaa acgtcgacgg tatcgataag cttgatatcg aattccgccc
     1801 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata
     1861 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg
     1921 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt
     1981 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag
     2041 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc
     2101 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga
     2161 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg
     2221 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat
     2281 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc
     2341 tttgaaaaac acgatgataa tatggccaca accatggtga gcaagggcga ggagctgttc
     2401 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     2461 gtgtctggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     2521 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     2581 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2641 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     2701 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2761 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     2821 aacgtctata tcatggccga caagcagaag aacggcatca aggcgaactt caagatccgc
     2881 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     2941 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     3001 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     3061 atcactctcg gcatggacga gctgtacaag taatgaatta attaagaatt cgataaaata
     3121 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg
     3181 caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag
     3241 aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg
     3301 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc
     3361 gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat
     3421 gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt
     3481 ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga
     3541 tagactgcgt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact
     3601 tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg
     3661 gtctttcatg ggtaacagtt tcttgaagtt ggagaacaac attctgaggg taggagtcga
     3721 atattaagta atcctgactc aattagccac tgttttgaat ccacatactc caatactcct
     3781 gaaatagttc attatggaca gcgcagaaag agctggggag aattgtgaaa ttgttatccg
     3841 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     3901 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3961 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4021 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4081 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4141 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4201 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4261 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4321 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4381 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4441 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4501 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4561 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4621 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4681 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4741 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4801 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4861 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4921 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4981 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5041 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5101 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5161 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5221 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5281 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5341 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5401 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5461 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5521 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5581 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5641 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5701 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5761 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5821 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5881 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5941 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     6001 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     6061 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     6121 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     6181 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     6241 gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa
     6301 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     6361 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcg
     6421 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     6481 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     6541 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     6601 cgatgcgtcc ggcgtagagg cgattagtcc aatttgttaa agacaggata tcagtggtcc
     6661 aggctctagt tttgactcaa caatatcacc agctgaagcc tatagagtac gagccataga
     6721 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta
     6781 ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag
     6841 aatagagaag ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg
     6901 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg
     6961 cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac
     7021 ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg
     7081 cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt
     7141 cctccgatag actgcgtcgc ccgggtaccc gtattcccaa taaagcctct tgctgtttgc
     7201 atccgaatcg tggactcgct gatccttggg agggtctcct cagattgatt gactgcccac
     7261 ctcgggggtc tttcatttgg aggttccacc gagatttgga gacccctgcc cagggaccac
     7321 cgaccccccc gccgggaggt aagctggcca gcggtcgttt cgtgtctgtc tctgtctttg
     7381 tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct gtactagtta gctaactagc
     7441 tctgtatctg gcggacccgt ggtggaactg acgagttctg aacacccggc cgcaaccctg
     7501 ggagacgtcc cagggacttt gggggccgtt tttgtggccc gacctgagga agggagtcga
     7561 tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg agacgagaac ctaaaacagt
     7621 tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc
     7681 tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg
     7741 aaaattaggg ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg
     7801 tcgagcggat cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct
     7861 gctctgcaga atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag
     7921 acctcatcac ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc
     7981 aggtccccta catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc
     8041 cctttgtaca ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg
     8101 aacctcctcg ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag
     8161 gcgccggaat t
//
