LOCUS       sequence-382975-        4876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector that expresses the HIV-1 GAG protein 
            with FRB fused to it.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4876)
  AUTHORS   Bogaert A, Van de Steene T, Vuylsteke M, Eyckerman S, Gevaert K
  TITLE     A decoupled Virotrap approach to study the interactomes of 
            N-terminal proteoforms.
  JOURNAL   Methods Enzymol. 2023;684:253-287. doi: 10.1016/bs.mie.2023.02.003. 
            Epub 2023 Mar 10.
  PUBMED    37230591
REFERENCE   2  (bases 1 to 4876)
  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: "Methods 
            Enzymol."; date: "2023"; volume: "684:253-287. doi"; pages: " 
            10.1016/bs.mie.2023.02.003. Epub 2023 Mar 10"
COMMENT     Sequence Label: pMET7-GAG-FRB
FEATURES             Location/Qualifiers
     source          1..4876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      27..36
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             33..1532
                     /codon_start=1
                     /gene="gag"
                     /product="gag protein from human immunodeficiency virus 1"
                     /label=HIV-1 gag
                     /translation="MGARASVLSGGELDRWEKIRLRPGGKKKYKLKHIVWASRELERFA
                     VNPGLLETSEGCRQILGQLQPSLQTGSEELRSLYNTVATLYCVHQRIEIKDTKEALDKI
                     EEEQNKSKKKAQQAAADTGHSNQVSQNYPIVQNIQGQMVHQAISPRTLNAWVKVVEEKA
                     FSPEVIPMFSALSEGATPQDLNTMLNTVGGHQAAMQMLKETINEEAAEWDRVHPVHAGP
                     IAPGQMREPRGSDIAGTTSTLQEQIGWMTHNPPIPVGEIYKRWIILGLNKIVRMYSPTS
                     ILDIRQGPKEPFRDYVDRFYKTLRAEQASQEVKNWMTETLLVQNANPDCKTILKALGPG
                     ATLEEMMTACQGVGGPGHKARVLAEAMSQVTNPATIMIQKGNFRNQRKTVKCFNCGKEG
                     HIAKNCRAPRKKGCWKCGKEGHQMKDCTERQANFLGKIWPSHKGRPGNFLQSRPEPTAP
                     PEESFRFGEETTTPSQKQEPIDKELYPLASLRSLFGSDPSSQ"
     CDS             1632..1910
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     polyA_signal    2000..2134
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2037..2056)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2091..2110
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2173..2190)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2344..2932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2424..2443)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3103..3894,3895..3963))
                     /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     3726..3745
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3964..4068)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4108..4128)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(4113..4442,4443..4457,4458..4726)
                     /label=SR-alpha promoter
                     /note="hybrid promoter consisting of the SV40 early 
                     promoter plus part of the long terminal repeat of human 
                     T-lymphotrophic virus 1"
     rep_origin      4293..4428
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     LTR             4458..4726
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     intron          4756..4852
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
ORIGIN
        1 ctctaaaagc tgcggggccc gctagcgcca ccatgggtgc gagagcgtca gtattaagcg
       61 ggggagaatt agatcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
      121 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
      181 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
      241 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
      301 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
      361 aaagtaagaa aaaagcacag caagcagcag ctgacacagg acacagcaat caggtcagcc
      421 aaaattaccc tatagtgcag aacatccagg ggcaaatggt acatcaggcc atatcaccta
      481 gaactttaaa tgcatgggta aaagtagtag aagagaaggc tttcagccca gaagtgatac
      541 ccatgttttc agcattatca gaaggagcca ccccacaaga tttaaacacc atgctaaaca
      601 cagtgggggg acatcaagca gccatgcaaa tgttaaaaga gaccatcaat gaggaagctg
      661 cagaatggga tagagtgcat ccagtgcatg cagggcctat tgcaccaggc cagatgagag
      721 aaccaagggg aagtgacata gcaggaacta ctagtaccct tcaggaacaa ataggatgga
      781 tgacacataa tccacctatc ccagtaggag aaatctataa aagatggata atcctgggat
      841 taaataaaat agtaagaatg tatagcccta ccagcattct ggacataaga caaggaccaa
      901 aggaaccctt tagagactat gtagaccgat tctataaaac tctaagagcc gagcaagctt
      961 cacaagaggt aaaaaattgg atgacagaaa ccttgttggt ccaaaatgcg aacccagatt
     1021 gtaagactat tttaaaagca ttgggaccag gagcgacact agaagaaatg atgacagcat
     1081 gtcagggagt ggggggaccc ggccataaag caagagtttt ggctgaagca atgagccaag
     1141 taacaaatcc agctaccata atgatacaga aaggcaattt taggaaccaa agaaagactg
     1201 ttaagtgttt caattgtggc aaagaagggc acatagccaa aaattgcagg gcccctagga
     1261 aaaagggctg ttggaaatgt ggaaaggaag gacaccaaat gaaagattgt actgagagac
     1321 aggctaattt tttagggaag atctggcctt cccacaaggg aaggccaggg aattttcttc
     1381 agagcagacc agagccaaca gccccaccag aagagagctt caggtttggg gaagagacaa
     1441 caactccctc tcagaagcag gagccgatag acaaggaact gtatccttta gcttccctca
     1501 gatcactctt tggcagcgac ccctcgtcac aaggcgggtc gacgctgacc ctgatcttca
     1561 gagaggccgt gagcgaaatc ctggagacct ccagattcgc ctacctgtac gacgaattca
     1621 tggcttctag aatcctctgg catgagatgt ggcatgaagg cctggaagag gcatctcgtt
     1681 tgtactttgg ggaaaggaac gtgaaaggca tgtttgaggt gctggagccc ttgcatgcta
     1741 tgatggaacg gggcccccag actctgaagg aaacatcctt taatcaggcc tatggtcgag
     1801 atttaatgga ggcccaagag tggtgcagga agtacatgaa atcagggaat gtcaaggacc
     1861 tcctccaagc ctgggacctc tattatcatg tgttccgacg aatctcaaag actagttaag
     1921 cggccgcata atctagagaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     1981 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     2041 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2101 caaactcatc aatgtatctt atcatgtctg gatccccggg taccgagctc gaattaattc
     2161 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2221 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2281 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2341 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2401 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2461 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2521 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2581 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2641 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2701 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2761 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2821 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2881 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2941 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3001 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3061 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3121 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3181 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3241 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3301 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3361 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3421 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3481 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3541 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3601 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3661 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3721 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3781 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3841 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3901 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3961 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4021 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4081 aaaagtgcca cctgcaagct tggctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     4141 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4201 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4261 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4321 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4381 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4441 aaaagctcct cgatcgaggg gctcgcatct ctccttcacg cgcccgctgc cctacctgag
     4501 gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa
     4561 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc
     4621 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc
     4681 aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctctgaaaaa
     4741 ccagaaagtt aactggtaag tttagtcttt ttgtctttta tttcaggtcc cggatccggt
     4801 ggtggtgcaa atcaaagaac tgctcctcag tggatgttgc ctttacttct aggcctgtac
     4861 ggaagtgtta cttctg
//
