LOCUS       sequence-438537-        8768 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ZBTB44 (pLxIS) in mammalian cells by retroviral 
            transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8768)
  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 8768)
  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-ZBTB44
FEATURES             Location/Qualifiers
     source          1..8768
                     /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    2391..2957
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2407..2959
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2551..2568)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2778..2797
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2971..2973,2974..2976,2977..3690)
                     /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(3016..3037)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3277..3296)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3624..3645
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3752..4266
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4428..4444
                     /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(4452..4458,4459..4476,4477..4482))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4497..4518
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4635..4652)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4806..5394)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4886..4905)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5565..6356,6357..6425))
                     /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     6188..6207
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6426..6530)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6598..6616
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6654..6676)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6776..6795
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6989..7011
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7139..7158
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             7358..7874
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7938..8279
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             8346..8762
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8692..8714
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8730..8746
                     /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 tgatgctgga caagaaaata gctccaactg caattttact
     1441 tctcgagatg gaagcatttc tccagtgtct tcagagtgca gtgctgtgga gaggaccatt
     1501 cctgtctgcc gagagtcccg gaggaagcgc aaaagctaca ttgttatgtc tcccgagagt
     1561 cccgtgaaat gtagcaccca aacaagttcc ccccaggtac tgaattcttc agcttcctat
     1621 gcagaaaata gaagtcagcc agttgactct tctttagctt ttccttggac tttccctttt
     1681 ggaatagatc gaaggattca gcctgagaaa gcaaagcagg cagaaaatac ccggacttta
     1741 gaattgcctg gcccatccga ggcaggtaga agagtggctg actatgtgac ttgtgagagc
     1801 acaaaaccca ccttgcctct aggtacagaa gaagatgtcc gggtcaaagt agaaagatta
     1861 agtgatgagg aggtccatga ggaagtgtcc cagcctgtta gtgcatctca gagctcactg
     1921 agtgatcagc agacagtgcc cggaagtgaa ccagttcaag aggaccttct gatcagtcca
     1981 cagtcttcct ctataggctc agtagatgaa ggagtcactg aaggattgcc tacacttcag
     2041 agcacctcta gcaccaatgc tcatgcagat gacgatgaca gattggaaaa tgttcagtat
     2101 ccctatcaac tctacattgc tccttctacc agcagtacag agcggccaag tccaaatggt
     2161 ccagacagac cgtttcagtg tccaacctgt ggggtccgct tcacccgtat tcagaatcta
     2221 aaacagcaca tgcttatcca ctcaggaatt aaaccatttc agtgtgactg ctgtggaaaa
     2281 aagtttacca gggcttactc gctaaagatg catcgcctaa agcacgaagt gatttcttga
     2341 taagcggccg cgtttaaacg tcgacggtat cgataagctt gatatcgaat tccgcccccc
     2401 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
     2461 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
     2521 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
     2581 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
     2641 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
     2701 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
     2761 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
     2821 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg
     2881 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     2941 gaaaaacacg atgataatat ggccacaacc atggtgagca agggcgagga gctgttcacc
     3001 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     3061 tctggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     3121 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     3181 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     3241 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     3301 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     3361 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     3421 gtctatatca tggccgacaa gcagaagaac ggcatcaagg cgaacttcaa gatccgccac
     3481 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     3541 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     3601 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     3661 actctcggca tggacgagct gtacaagtaa tgaattaatt aagaattcga taaaataaaa
     3721 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     3781 gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag
     3841 ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg
     3901 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc
     3961 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
     4021 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
     4081 ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag
     4141 actgcgtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt
     4201 ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc
     4261 tttcatgggt aacagtttct tgaagttgga gaacaacatt ctgagggtag gagtcgaata
     4321 ttaagtaatc ctgactcaat tagccactgt tttgaatcca catactccaa tactcctgaa
     4381 atagttcatt atggacagcg cagaaagagc tggggagaat tgtgaaattg ttatccgctc
     4441 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4501 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4561 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4621 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4681 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4741 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4801 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4861 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4921 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4981 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5041 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5101 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5161 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5221 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5281 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5341 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5401 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5461 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5521 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5581 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5641 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5701 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5761 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5821 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5881 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5941 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6001 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6061 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6121 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6181 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6241 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6301 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6361 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6421 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6481 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6541 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     6601 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     6661 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     6721 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     6781 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     6841 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     6901 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     6961 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggcgcaa
     7021 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     7081 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     7141 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     7201 tgcgtccggc gtagaggcga ttagtccaat ttgttaaaga caggatatca gtggtccagg
     7261 ctctagtttt gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagataa
     7321 aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt
     7381 ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat
     7441 agagaagttc agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata
     7501 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg
     7561 tcccgccctc agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg
     7621 aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc
     7681 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct
     7741 ccgatagact gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc
     7801 cgaatcgtgg actcgctgat ccttgggagg gtctcctcag attgattgac tgcccacctc
     7861 gggggtcttt catttggagg ttccaccgag atttggagac ccctgcccag ggaccaccga
     7921 cccccccgcc gggaggtaag ctggccagcg gtcgtttcgt gtctgtctct gtctttgtgc
     7981 gtgtttgtgc cggcatctaa tgtttgcgcc tgcgtctgta ctagttagct aactagctct
     8041 gtatctggcg gacccgtggt ggaactgacg agttctgaac acccggccgc aaccctggga
     8101 gacgtcccag ggactttggg ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt
     8161 ggaatccgac cccgtcagga tatgtggttc tggtaggaga cgagaaccta aaacagttcc
     8221 cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc
     8281 agcgctgcag catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa
     8341 attagggcca gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg
     8401 agcggatcgc tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct
     8461 ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc
     8521 tcatcaccca ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg
     8581 tcccctacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct
     8641 ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac
     8701 ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg
     8761 ccggaatt
//
