LOCUS       sequence-403391-        5354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector that expresses HIV-1 GAG with EGFP fused
            to it.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5354)
  AUTHORS   Eyckerman S, Titeca K, Van Quickelberghe E, Cloots E, Verhee A, 
            Samyn N, De Ceuninck L, Timmerman E, De Sutter D, Lievens S, Van 
            Calenbergh S, Gevaert K, Tavernier J
  TITLE     Trapping mammalian protein complexes in viral particles.
  JOURNAL   Nat Commun. 2016 Apr 28;7:11416. doi: 10.1038/ncomms11416.
  PUBMED    27122307
REFERENCE   2  (bases 1 to 5354)
  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: "Nat 
            Commun."; date: "2016-04-28"; volume: "7:11416. doi"; pages: " 
            10.1038/ncomms11416"
COMMENT     Sequence Label: pMET7-GAG-EGFP
FEATURES             Location/Qualifiers
     source          1..5354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     337..357
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        397..501
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(502..570,571..1362)
                     /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     complement(720..739)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1533..2121
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2022..2041
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2275..2292
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    2331..2465
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2355..2374)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2409..2428
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(2547..3260,3261..3263,3264..3266))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(2550..3260,3261..3263,3264..3266))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2592..2613)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2941..2960
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3200..3221
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3263..3272
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(3339..4838)
                     /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"
     regulatory      4835..4844
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          4895..4991
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     LTR             5021..5289
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(join(5305..5354,1..267))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5319..5354,1..100)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg
       61 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga
      121 ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
      181 gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
      241 ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca
      301 tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac acattccaca
      361 gccaagcttg caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
      421 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
      481 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
      541 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
      601 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
      661 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
      721 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
      781 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
      841 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
      901 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
      961 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     1021 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     1081 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     1141 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     1201 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     1261 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     1321 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     1381 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     1441 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     1501 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     1561 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     1621 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     1681 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     1741 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     1801 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     1861 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     1921 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     1981 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2041 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2101 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     2161 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     2221 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     2281 tcagtgagcg aggaagcgga agaggaatta attcgagctc ggtacccggg gatccagaca
     2341 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     2401 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     2461 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     2521 ttttttctct agattatgcg gccgctttac ttgtacagct cgtccatgcc gagagtgatc
     2581 ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg
     2641 ctcagggcgg actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg
     2701 atgggggtgt tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg
     2761 cggatcttga agttcacctt gatgccgttc ttctgcttgt cggccatgat atagacgttg
     2821 tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg
     2881 atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg
     2941 gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc
     3001 atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc
     3061 acgccgtagg tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg
     3121 cagatgaact tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg
     3181 ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg
     3241 aacagctcct cgcccttgct caccatggtg gcgaccggcc ggtggatccc gggcccgcgg
     3301 taccgtcgac tgcagaattc tgagctcgtc gacccgcctt gtgacgaggg gtcgctgcca
     3361 aagagtgatc tgagggaagc taaaggatac agttccttgt ctatcggctc ctgcttctga
     3421 gagggagttg ttgtctcttc cccaaacctg aagctctctt ctggtggggc tgttggctct
     3481 ggtctgctct gaagaaaatt ccctggcctt cccttgtggg aaggccagat cttccctaaa
     3541 aaattagcct gtctctcagt acaatctttc atttggtgtc cttcctttcc acatttccaa
     3601 cagccctttt tcctaggggc cctgcaattt ttggctatgt gcccttcttt gccacaattg
     3661 aaacacttaa cagtctttct ttggttccta aaattgcctt tctgtatcat tatggtagct
     3721 ggatttgtta cttggctcat tgcttcagcc aaaactcttg ctttatggcc gggtcccccc
     3781 actccctgac atgctgtcat catttcttct agtgtcgctc ctggtcccaa tgcttttaaa
     3841 atagtcttac aatctgggtt cgcattttgg accaacaagg tttctgtcat ccaatttttt
     3901 acctcttgtg aagcttgctc ggctcttaga gttttataga atcggtctac atagtctcta
     3961 aagggttcct ttggtccttg tcttatgtcc agaatgctgg tagggctata cattcttact
     4021 attttattta atcccaggat tatccatctt ttatagattt ctcctactgg gataggtgga
     4081 ttatgtgtca tccatcctat ttgttcctga agggtactag tagttcctgc tatgtcactt
     4141 ccccttggtt ctctcatctg gcctggtgca ataggccctg catgcactgg atgcactcta
     4201 tcccattctg cagcttcctc attgatggtc tcttttaaca tttgcatggc tgcttgatgt
     4261 ccccccactg tgtttagcat ggtgtttaaa tcttgtgggg tggctccttc tgataatgct
     4321 gaaaacatgg gtatcacttc tgggctgaaa gccttctctt ctactacttt tacccatgca
     4381 tttaaagttc taggtgatat ggcctgatgt accatttgcc cctggatgtt ctgcactata
     4441 gggtaatttt ggctgacctg attgctgtgt cctgtgtcag ctgctgcttg ctgtgctttt
     4501 ttcttacttt tgttttgctc ttcctctatc ttgtctaaag cttccttggt gtcttttatc
     4561 tctatccttt gatgcacaca atagagggtt gctactgtat tatataatga tctaagttct
     4621 tctgatcctg tctgaaggga tggttgtagc tgtcccagta tttgtctaca gccttctgat
     4681 gtttctaaca ggccaggatt aactgcgaat cgttctagct ccctgcttgc ccatactata
     4741 tgttttaatt tatatttttt ctttccccct ggccttaacc gaattttttc ccatcgatct
     4801 aattctcccc cgcttaatac tgacgctctc gcacccatgg tggcgctagc gggccccgca
     4861 gcttttagag cagaagtaac acttccgtac aggcctagaa gtaaaggcaa catccactga
     4921 ggagcagttc tttgatttgc accaccaccg gatccgggac ctgaaataaa agacaaaaag
     4981 actaaactta ccagttaact ttctggtttt tcagagcttg gatctgtaac ggcgcagaac
     5041 agaaaacgaa acaaagacgt agagttgagc aagcagggtc aggcaaagcg tggagagccg
     5101 gctgagtcta ggtaggctcc aagggagcgc cggacaaagg cccggtctcg acctgagctt
     5161 taaacttacc tagacggcgg acgcagttca ggaggcacca caggcgggag gcggcagaac
     5221 gcgactcaac cggcgtggat ggcggcctca ggtagggcag cgggcgcgtg aaggagagat
     5281 gcgagcccct cgatcgagga gctttttgca aaagcctagg cctccaaaaa agcctcctca
     5341 ctacttctgg aata
//
