LOCUS       sequence-438525-        8252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Myoglobin (pLxIS-PLPLR) in mammalian cells by 
            retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8252)
  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 8252)
  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-Myoglobin(pLxIS-PLPLR)
FEATURES             Location/Qualifiers
     source          1..8252
                     /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    1875..2441
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1891..2443
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2035..2052)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2262..2281
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2455..2457,2458..2460,2461..3174)
                     /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(2500..2521)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2761..2780)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3108..3129
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3236..3750
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3912..3928
                     /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(3936..3942,3943..3960,3961..3966))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3981..4002
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4119..4136)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4290..4878)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4370..4389)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5049..5840,5841..5909))
                     /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     5672..5691
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5910..6014)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6082..6100
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6138..6160)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6260..6279
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6473..6495
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6623..6642
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6842..7358
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7422..7763
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             7830..8246
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8176..8198
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8214..8230
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 agatctgcca ccatggacta caaagacgac gatgataaag gagttcaggt ggagactatt
       61 tctcctggag atggccgcac tttcccaaaa cgcggtcaga catgcgtggt gcattacacc
      121 ggcatgctgg aggacggcaa gaaggtggac tcttctagag acagaaataa gccctttaaa
      181 ttcatgcttg gaaagcaaga agtgatccgc ggttgggaag agggtgtcgc ccagatgagc
      241 gtggggcagc gcgccaagtt gaccatttct cccgactatg cttacggtgc aacaggccat
      301 cccggaataa ttcctcctca tgcaacactc gtctttgacg ttgaactttt gaagctggag
      361 actcgggggg tacaggttga aacaatctca ccaggtgatg ggcgcacttt tccaaaaaga
      421 ggacaaacat gcgtggtaca ctacactgga atgttggagg atggtaagaa agtggactct
      481 agtagagatc ggaataagcc atttaagttt atgcttggga aacaagaggt aattcgcggc
      541 tgggaggaag gggtagccca aatgagtgtt ggccaacgcg ccaaactgac tatcagtcca
      601 gactatgcct atggagccac cggacatcca ggcattattc ctcctcacgc cacccttgtc
      661 tttgatgtag agcttctgaa gttggagact agaggggtac aagtggagac aatatccccc
      721 ggtgacggac gaacatttcc taagcgcgga caaacttgcg tggtgcacta tacaggtatg
      781 ctggaggatg gaaaaaaggt tgatagctca cgcgaccgaa acaaaccttt taaattcatg
      841 ctcggtaaac aggaagtcat tcgcgggtgg gaggaaggtg tggctcagat gtccgtgggt
      901 cagcgggcta aattgacaat aagccccgat tatgcatacg gggccaccgg tcatcctgga
      961 atcataccac ctcatgcaac acttgtattc gatgtcgaac tgctcaagct cgagacacgg
     1021 ggagtgcagg tagagacaat ttccccaggc gacgggagga cttttcctaa aaggggccaa
     1081 acttgtgtcg tccactacac cggaatgctt gaagatggca aaaaagtaga tagctcaaga
     1141 gacaggaaca aaccattcaa gttcatgctg ggtaagcaag aagttattcg agggtgggaa
     1201 gagggcgtag cccagatgtc tgttggtcaa agagctaagc tgaccatttc acccgattac
     1261 gcctatggcg ccactggtca ccctggtatt atacctcccc acgctacact cgtgttcgac
     1321 gtcgagctcc tcaagctcga aggggctggc gctggagcca tggggctcag cgacggcgag
     1381 tggcagcttg tccttaatgt atgggggaaa gtgaatctgg tgatttcagg gcatggccag
     1441 gaagtcttga taggtttgtt caaacccctg cctctgcgac tggataagtt tgataagttc
     1501 aaaaatctta aatcagaaga ggatatgaaa ggttctgagg atttgaagaa acacggatgc
     1561 acagtattga cagctctggg taccattctc aagaaaaagg gccaacacgc tgcagaaata
     1621 caacccctgg cacagtcaca tgctacaaaa cataaaattc ctgttaaata tctcgagttt
     1681 atctcagaga ttattatcga ggtgttgaag aagcggcact ctggggactt tggtgccgat
     1741 gctcaggggg ctatgagtaa ggccttggag ctcttccgaa acgatatagc agctaagtac
     1801 aaagaattgg gcttccaggg ctgataagcg gccgcgttta aacgtcgacg gtatcgataa
     1861 gcttgatatc gaattccgcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     1921 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     1981 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     2041 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     2101 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     2161 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     2221 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     2281 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     2341 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     2401 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtg
     2461 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2521 gtaaacggcc acaagttcag cgtgtctggc gagggcgagg gcgatgccac ctacggcaag
     2581 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2641 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     2701 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2761 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2821 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2881 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     2941 aaggcgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     3001 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     3061 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     3121 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaatgaatt
     3181 aattaagaat tcgataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa
     3241 agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa
     3301 aatacataac tgagaataga gaagttcaga tcaaggttag gaacagagag acagcagaat
     3361 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     3421 atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag atgtttccag
     3481 ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt
     3541 ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca
     3601 ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtgtat ccaataaacc
     3661 ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg
     3721 attgactacc cgtcagcggg ggtctttcat gggtaacagt ttcttgaagt tggagaacaa
     3781 cattctgagg gtaggagtcg aatattaagt aatcctgact caattagcca ctgttttgaa
     3841 tccacatact ccaatactcc tgaaatagtt cattatggac agcgcagaaa gagctgggga
     3901 gaattgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3961 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4021 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4081 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4141 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4201 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4261 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4321 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4381 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4441 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4501 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4561 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4621 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4681 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4741 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4801 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4861 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4921 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4981 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5041 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5101 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5161 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5221 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5281 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5341 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5401 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5461 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5521 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5581 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5641 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5701 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5761 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5821 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5881 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5941 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6001 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     6061 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     6121 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     6181 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     6241 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     6301 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     6361 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     6421 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     6481 cgacgttgta aaacgacggc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     6541 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     6601 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     6661 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gcgattagtc caatttgtta
     6721 aagacaggat atcagtggtc caggctctag ttttgactca acaatatcac cagctgaagc
     6781 ctatagagta cgagccatag ataaaataaa agattttatt tagtctccag aaaaaggggg
     6841 gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg
     6901 catggaaaat acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca
     6961 gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     7021 agaacagatg gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg
     7081 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     7141 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     7201 cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtattccca
     7261 ataaagcctc ttgctgtttg catccgaatc gtggactcgc tgatccttgg gagggtctcc
     7321 tcagattgat tgactgccca cctcgggggt ctttcatttg gaggttccac cgagatttgg
     7381 agacccctgc ccagggacca ccgacccccc cgccgggagg taagctggcc agcggtcgtt
     7441 tcgtgtctgt ctctgtcttt gtgcgtgttt gtgccggcat ctaatgtttg cgcctgcgtc
     7501 tgtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttct
     7561 gaacacccgg ccgcaaccct gggagacgtc ccagggactt tgggggccgt ttttgtggcc
     7621 cgacctgagg aagggagtcg atgtggaatc cgaccccgtc aggatatgtg gttctggtag
     7681 gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc
     7741 gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt ctctgtctga
     7801 ctgtgtttct gtatttgtct gaaaattagg gccagactgt taccactccc ttaagtttga
     7861 ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga
     7921 agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc ggatggccgc
     7981 gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc ttttcacctg
     8041 gcccgcatgg acacccagac caggtcccct acatcgtgac ctgggaagcc ttggcttttg
     8101 acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct cttcctccat
     8161 ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc tccctttatc
     8221 cagccctcac tccttctcta ggcgccggaa tt
//
