LOCUS       sequence-388782-        7032 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  a bicistronic IRES-GFP containing vector which is a variant of pMIG 
            that contains a posttranscriptional regulatory element from the 
            woodchuck hepatitis virus .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7032)
  AUTHORS   Refaeli Y, Van Parijs L, Alexander SI, Abbas AK
  TITLE     Interferon gamma is required for activation-induced death of T 
            lymphocytes.
  JOURNAL   J Exp Med. 2002 Oct 7. 196(7):999-1005.
  PUBMED    12370261
REFERENCE   2  (bases 1 to 7032)
  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"; journalName: "J Exp Med. 
            2002 Oct 7. 196(7):999-1005."
COMMENT     Sequence Label: pMIG-w
FEATURES             Location/Qualifiers
     source          1..7032
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    15..567
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(159..176)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     386..405
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(579..581,582..584,585..1298)
                     /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(624..645)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(885..904)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1232..1253
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1408..1996
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1461..1481)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1879..1890)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2043..2557
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    2719..2735
                     /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(2743..2749,2750..2767,2768..2773))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2788..2809
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2926..2943)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3097..3685)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3177..3196)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3856..4647,4648..4716))
                     /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     4479..4498
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4717..4821)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4889..4907
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4945..4967)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5067..5086
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5280..5302
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5430..5449
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             5649..6165
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6229..6570
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     primer_bind     6983..7005
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
ORIGIN
        1 sycccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
       61 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
      121 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
      181 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc
      241 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
      301 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
      361 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
      421 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt
      481 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
      541 tttcctttga aaaacacgat gataatatgg ccacaaccat ggtgagcaag ggcgaggagc
      601 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      661 tcagcgtgtc tggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
      721 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
      781 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      841 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
      901 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
      961 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1021 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggcg aacttcaaga
     1081 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1141 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     1201 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1261 ccgggatcac tctcggcatg gacgagctgt acaagtaatg aattaattaa gaattatcac
     1321 cgcttctatt cagccagtaa ggcctgtctt taatggcctc cggcatgaga cactttctag
     1381 agtcgacctg cagccaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga
     1441 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     1501 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     1561 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     1621 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     1681 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     1741 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     1801 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     1861 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     1921 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     1981 tgggccgcct ccccgcatcg ataaaataaa agattttatt tagtctccag aaaaaggggg
     2041 gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg
     2101 catggaaaat acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca
     2161 gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     2221 agaacagatg gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg
     2281 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     2341 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     2401 cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtgtatcca
     2461 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
     2521 ctgagtgatt gactacccgt cagcgggggt ctttcatggg taacagtttc ttgaagttgg
     2581 agaacaacat tctgagggta ggagtcgaat attaagtaat cctgactcaa ttagccactg
     2641 ttttgaatcc acatactcca atactcctga aatagttcat tatggacagc gcagaaagag
     2701 ctggggagaa ttgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2761 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2821 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2881 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2941 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3001 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3061 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3121 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3181 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3241 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3301 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3361 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3421 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3481 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3541 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3601 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3661 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3721 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3781 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3841 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3901 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3961 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4021 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4081 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4141 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4201 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4261 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4321 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4381 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4441 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4501 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4561 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4621 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4681 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4741 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4801 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     4861 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     4921 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     4981 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     5041 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     5101 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     5161 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     5221 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     5281 ccagtcacga cgttgtaaaa cgacggcgca aggaatggtg catgcaagga gatggcgccc
     5341 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     5401 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     5461 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggcg attagtccaa
     5521 tttgttaaag acaggatatc agtggtccag gctctagttt tgactcaaca atatcaccag
     5581 ctgaagccta tagagtacga gccatagata aaataaaaga ttttatttag tctccagaaa
     5641 aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt
     5701 tgcaaggcat ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag
     5761 agagacagca gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     5821 agggccaaga acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca
     5881 tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac
     5941 caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag
     6001 cccacaaccc ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt
     6061 attcccaata aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag
     6121 ggtctcctca gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga
     6181 gatttggaga cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc
     6241 ggtcgtttcg tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc
     6301 ctgcgtctgt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
     6361 gagttctgaa cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt
     6421 tgtggcccga cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt
     6481 ctggtaggag acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt
     6541 tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc
     6601 tgtctgactg tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta
     6661 agtttgacct taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat
     6721 gtcaagaaga gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga
     6781 tggccgcgag acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt
     6841 tcacctggcc cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg
     6901 gcttttgacc cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt
     6961 cctccatccg ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc
     7021 ctttatccag cc
//
