LOCUS       sequence-439086-        8040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GMIP N-terminal truncation 1 (16-84) in mammalian 
            cells by retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8040)
  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 8040)
  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-GMIP(trunc N1)
FEATURES             Location/Qualifiers
     source          1..8040
                     /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    1662..2228
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1678..2230
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1822..1839)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2049..2068
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2242..2244,2245..2247,2248..2961)
                     /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(2287..2308)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2548..2567)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2895..2916
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3023..3538
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3700..3716
                     /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(3724..3730,3731..3748,3749..3754))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3769..3790
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3907..3924)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4078..4666)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4158..4177)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4837..5628,5629..5697))
                     /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     5460..5479
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5698..5802)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5870..5888
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5926..5948)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6048..6067
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6261..6283
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6411..6430
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6630..7146
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7210..7551
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             7618..8034
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7964..7986
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8002..8018
                     /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 taagagatac agtgacatct tccagagcct ggacaacctc
     1441 gagatatcat tggggaacgt aacttttgat cccctggctg gagaccctgt acgcagagaa
     1501 gacctggagc ctgacaaagc tgacacagcc acagtggtga ctgaggagaa cagtgaagcc
     1561 agtagttggc gggatctctc cccagaaggt cctgcacccc tcacagagtg ataagcggcc
     1621 gcgtttaaac gtcgacggta tcgataagct tgatatcgaa ttccgccccc ccccctaacg
     1681 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     1741 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     1801 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     1861 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     1921 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     1981 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     2041 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     2101 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     2161 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     2221 gatgataata tggccacaac catggtgagc aagggcgagg agctgttcac cggggtggtg
     2281 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtctggcgag
     2341 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2401 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     2461 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2521 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2581 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2641 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2701 atggccgaca agcagaagaa cggcatcaag gcgaacttca agatccgcca caacatcgag
     2761 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2821 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     2881 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2941 atggacgagc tgtacaagta atgaattaat taagaattcg ataaaataaa agattttatt
     3001 tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta
     3061 agtaacgcca ttttgcaagg catggaaaat acataactga gaatagagaa gttcagatca
     3121 aggttaggaa cagagagaca gcagaatatg ggccaaacag gatatctgtg gtaagcagtt
     3181 cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtcccgc cctcagcagt
     3241 ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
     3301 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
     3361 ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgata gactgcgtcg
     3421 cccgggtacc cgtattccca ataaagcctc ttgctgtttg catccgactt gtggtctcgc
     3481 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcatgg
     3541 gtaacagttt cttgaagttg gagaacaaca ttctgagggt aggagtcgaa tattaagtaa
     3601 tcctgactca attagccact gttttgaatc cacatactcc aatactcctg aaatagttca
     3661 ttatggacag cgcagaaaga gctggggaga attgtgaaat tgttatccgc tcacaattcc
     3721 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3781 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3841 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3901 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3961 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4021 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4081 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4141 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4201 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4261 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4321 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4381 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4441 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4501 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4561 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4621 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4681 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4741 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4801 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4861 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4921 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4981 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5041 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5101 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5161 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5221 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5281 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5341 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5401 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5461 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5521 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5581 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5641 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5701 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5761 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5821 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     5881 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     5941 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     6001 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     6061 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     6121 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     6181 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     6241 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcgc aaggaatggt
     6301 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     6361 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     6421 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     6481 gcgtagaggc gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt
     6541 ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag
     6601 attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag
     6661 ctagcttaag taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt
     6721 tcagatcaag gttaggaaca gagagacagc agaatatggg ccaaacagga tatctgtggt
     6781 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtcccgccc
     6841 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
     6901 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
     6961 tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgataga
     7021 ctgcgtcgcc cgggtacccg tattcccaat aaagcctctt gctgtttgca tccgaatcgt
     7081 ggactcgctg atccttggga gggtctcctc agattgattg actgcccacc tcgggggtct
     7141 ttcatttgga ggttccaccg agatttggag acccctgccc agggaccacc gacccccccg
     7201 ccgggaggta agctggccag cggtcgtttc gtgtctgtct ctgtctttgt gcgtgtttgt
     7261 gccggcatct aatgtttgcg cctgcgtctg tactagttag ctaactagct ctgtatctgg
     7321 cggacccgtg gtggaactga cgagttctga acacccggcc gcaaccctgg gagacgtccc
     7381 agggactttg ggggccgttt ttgtggcccg acctgaggaa gggagtcgat gtggaatccg
     7441 accccgtcag gatatgtggt tctggtagga gacgagaacc taaaacagtt cccgcctccg
     7501 tctgaatttt tgctttcggt ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc
     7561 agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattagggc
     7621 cagactgtta ccactccctt aagtttgacc ttaggtcact ggaaagatgt cgagcggatc
     7681 gctcacaacc agtcggtaga tgtcaagaag agacgttggg ttaccttctg ctctgcagaa
     7741 tggccaacct ttaacgtcgg atggccgcga gacggcacct ttaaccgaga cctcatcacc
     7801 caggttaaga tcaaggtctt ttcacctggc ccgcatggac acccagacca ggtcccctac
     7861 atcgtgacct gggaagcctt ggcttttgac ccccctccct gggtcaagcc ctttgtacac
     7921 cctaagcctc cgcctcctct tcctccatcc gccccgtctc tcccccttga acctcctcgt
     7981 tcgaccccgc ctcgatcctc cctttatcca gccctcactc cttctctagg cgccggaatt
//
