LOCUS       sequence-438538-        8381 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ZC3H12B (pLxIS) in mammalian cells by retroviral 
            transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8381)
  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 8381)
  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-ZC3H12B
FEATURES             Location/Qualifiers
     source          1..8381
                     /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    2004..2570
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2020..2572
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2164..2181)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2391..2410
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2584..2586,2587..2589,2590..3303)
                     /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(2629..2650)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2890..2909)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3237..3258
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3365..3879
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4041..4057
                     /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(4065..4071,4072..4089,4090..4095))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4110..4131
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4248..4265)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4419..5007)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4499..4518)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5178..5969,5970..6038))
                     /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     5801..5820
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6039..6143)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6211..6229
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6267..6289)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6389..6408
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6602..6624
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6752..6771
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6971..7487
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7551..7892
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             7959..8375
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8305..8327
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8343..8359
                     /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 tagaaagaga cctgtggttc ctgagcataa gaagcagcca
     1441 tgtccttatg gcaaaaaatg tacctacggc cacaagtgca aatactatca tccggagcgg
     1501 gccaaccaac cccagcgttc ggtggctgat gagctccgca tcagtgccaa actgtccaca
     1561 gtgaaaataa tgagtgaaga caccctggcc aaatgtggcg cagggatgtc tactgccaaa
     1621 ggtgagataa cctcagagat aaaacgtgtg gcctccaagc gccaatcaga tcccagcatc
     1681 cggtctgtgg ctgtagagcc tgaggagtgg ctgtccatcg cccgtaaacc tgaggccagt
     1741 tctgtccctt cacttgtgac cgctttaagt gtccctacaa tccctcctcc caaaagccat
     1801 gcagtgggtg cactcaacac ccgttcagcc agcagcccag tgccagggtc ctcccatttt
     1861 ccccatcaaa aggcctcttt ggaacacatg gtcagcatgc aatatccacc tatcctggtt
     1921 accaacagcc atggcacccc tattaactat tgataagcgg ccgcgtttaa acgtcgacgg
     1981 tatcgataag cttgatatcg aattccgccc ccccccctaa cgttactggc cgaagccgct
     2041 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     2101 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     2161 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     2221 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     2281 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     2341 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     2401 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     2461 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
     2521 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca
     2581 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     2641 gacggcgacg taaacggcca caagttcagc gtgtctggcg agggcgaggg cgatgccacc
     2701 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     2761 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     2821 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2881 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2941 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     3001 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     3061 aacggcatca aggcgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     3121 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     3181 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     3241 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     3301 taatgaatta attaagaatt cgataaaata aaagatttta tttagtctcc agaaaaaggg
     3361 gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa
     3421 ggcatggaaa atacataact gagaatagag aagttcagat caaggttagg aacagagaga
     3481 cagcagaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
     3541 caagaacaga tggtccccag atgcggtccc gccctcagca gtttctagag aaccatcaga
     3601 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc
     3661 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
     3721 aacccctcac tcggcgcgcc agtcctccga tagactgcgt cgcccgggta cccgtgtatc
     3781 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
     3841 ctctgagtga ttgactaccc gtcagcgggg gtctttcatg ggtaacagtt tcttgaagtt
     3901 ggagaacaac attctgaggg taggagtcga atattaagta atcctgactc aattagccac
     3961 tgttttgaat ccacatactc caatactcct gaaatagttc attatggaca gcgcagaaag
     4021 agctggggag aattgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4081 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4141 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4201 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4261 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4321 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4381 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4441 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4501 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4561 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4621 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4681 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4741 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4801 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4861 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4921 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4981 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5041 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5101 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5161 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5221 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5281 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5341 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5401 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5461 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5521 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5581 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5641 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5701 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5761 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5821 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5881 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5941 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6001 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6061 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6121 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     6181 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     6241 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     6301 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     6361 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     6421 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     6481 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     6541 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     6601 tcccagtcac gacgttgtaa aacgacggcg caaggaatgg tgcatgcaag gagatggcgc
     6661 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     6721 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     6781 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg cgattagtcc
     6841 aatttgttaa agacaggata tcagtggtcc aggctctagt tttgactcaa caatatcacc
     6901 agctgaagcc tatagagtac gagccataga taaaataaaa gattttattt agtctccaga
     6961 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat
     7021 tttgcaaggc atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac
     7081 agagagacag cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
     7141 tcagggccaa gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac
     7201 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
     7261 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
     7321 agcccacaac ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc
     7381 gtattcccaa taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg
     7441 agggtctcct cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc
     7501 gagatttgga gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca
     7561 gcggtcgttt cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc
     7621 gcctgcgtct gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg
     7681 acgagttctg aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt
     7741 tttgtggccc gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg
     7801 ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg
     7861 tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc
     7921 tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct
     7981 taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag
     8041 atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg
     8101 gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct
     8161 tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct
     8221 tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc
     8281 ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct
     8341 ccctttatcc agccctcact ccttctctag gcgccggaat t
//
