LOCUS       sequence-96382-e        6465 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression plasmid; drives constitutive, low-level 
            expression from retroviral LTR; retroviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6465)
  AUTHORS   Kofoed EM, Vance RE
  TITLE     Innate immune recognition of bacterial ligands by NAIPs determines 
            inflammasome specificity.
  JOURNAL   Nature. 2011 Aug 28;477(7366):592-5. doi: 10.1038/nature10394.
  PUBMED    21874021
REFERENCE   2  (bases 1 to 6465)
  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"; journalName: 
            "Nature"; date: "2011-08-28- 28"; volume: "477"; issue: "7366"; 
            pages: "592-5"
COMMENT     Sequence Label: mscv2.2
FEATURES             Location/Qualifiers
     source          1..6465
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..80
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     96..112
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    200..766
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    216..768
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(360..377)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     587..606
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(780..782,783..785,786..1499)
                     /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(825..846)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1086..1105)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1433..1454
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             1552..2066
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(2235..2251)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2235..2251)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2248..2270)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2259..2275
                     /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(2283..2289,2290..2307,2308..2313))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2328..2349
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2466..2483)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2637..3225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2717..2736)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3396..4187,4188..4256))
                     /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     4019..4038
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4257..4361)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4429..4447
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4485..4507)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4607..4626
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4820..4842
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4970..4989
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             5189..5705
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    5769..6110
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(6177..6465,1..128)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
       61 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
      121 taggcgccgg aattagatct ctcgagatcg atgcggccgc gtttaaacgt cgacggtatc
      181 gataagcttg atatcgaatt ccgccccccc ccctaacgtt actggccgaa gccgcttgga
      241 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
      301 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
      361 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
      421 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
      481 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
      541 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
      601 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
      661 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
      721 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaacca
      781 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
      841 gcgacgtaaa cggccacaag ttcagcgtgt ctggcgaggg cgagggcgat gccacctacg
      901 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
      961 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     1021 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     1081 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     1141 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     1201 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     1261 gcatcaaggc gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     1321 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     1381 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     1441 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtgat
     1501 aagaattcga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
     1561 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaata
     1621 cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag cagaatatgg
     1681 gccaaacagg atatctgtgg taagcagttc ctgcccccgg ctcagggcca agaacagatg
     1741 gtccccgatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
     1801 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg
     1861 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg
     1921 gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtgtatccaa taaaccctct
     1981 tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
     2041 actacccgtc agcgggggtc tttcatgggt aacagtttct tgaagttgga gaacaacatt
     2101 ctgagggtag gagtcgaata ttaagtaatc ctgactcaat tagccactgt tttgaatcca
     2161 catactccaa tactcctgaa atagttcatt atggacagcg cagaagagct ggggagaatt
     2221 aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     2281 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     2341 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     2401 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2461 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2521 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2581 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2641 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2701 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2761 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2821 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2881 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2941 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3001 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3061 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3121 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3181 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3241 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3301 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3361 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3421 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3481 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3541 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3601 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3661 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3721 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3781 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3841 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3901 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     3961 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4021 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4081 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4141 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4201 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4261 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4321 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4381 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     4441 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     4501 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     4561 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     4621 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     4681 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     4741 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     4801 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggcgca aggaatggtg
     4861 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     4921 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     4981 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     5041 cgtagaggcg attagtccaa tttgttaaag acaggatatc agtggtccag gctctagttt
     5101 tgactcaaca atatcaccag ctgaagccta tagagtacga gccatagata aaataaaaga
     5161 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc
     5221 tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa tagagaagtt
     5281 cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat atctgtggta
     5341 agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg gtcccgccct
     5401 cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc
     5461 tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct
     5521 ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc tccgatagac
     5581 tgcgtcgccc gggtacccgt attcccaata aagcctcttg ctgtttgcat ccgaatcgtg
     5641 gactcgctga tccttgggag ggtctcctca gattgattga ctgcccacct cgggggtctt
     5701 tcatttggag gttccaccga gatttggaga cccctgccca gggaccaccg acccccccgc
     5761 cgggaggtaa gctggccagc ggtcgtttcg tgtctgtctc tgtctttgtg cgtgtttgtg
     5821 ccggcatcta atgtttgcgc ctgcgtctgt actagttagc taactagctc tgtatctggc
     5881 ggacccgtgg tggaactgac gagttctgaa cacccggccg caaccctggg agacgtccca
     5941 gggactttgg gggccgtttt tgtggcccga cctgaggaag ggagtcgatg tggaatccga
     6001 ccccgtcagg atatgtggtt ctggtaggag acgagaacct aaaacagttc ccgcctccgt
     6061 ctgaattttt gctttcggtt tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca
     6121 gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagggcc
     6181 agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
     6241 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
     6301 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
     6361 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
     6421 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtca
//
