LOCUS       sequence-382463-        4984 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of the eDHFR control bait in Virotrap.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4984)
  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 4984)
  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-eDHFR
FEATURES             Location/Qualifiers
     source          1..4984
                     /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             1548..2021
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="ISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRHT
                     WESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLP
                     KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     polyA_signal    2108..2242
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2145..2164)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2199..2218
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2281..2298)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2452..3040)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2532..2551)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3211..4002,4003..4071))
                     /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     3834..3853
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4072..4176)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4216..4236)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(4221..4550,4551..4565,4566..4834)
                     /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      4401..4536
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     LTR             4566..4834
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     intron          4864..4960
                     /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 gagcggaatc agtctgattg
     1561 cggcgttagc ggtagatcgc gttatcggca tggaaaacgc catgccgtgg aacctgcctg
     1621 ccgatctcgc ctggtttaaa cgcaacacct taaataaacc cgtgattatg ggccgccata
     1681 cctgggaatc aatcggtcgt ccgttgccag gacgcaaaaa tattatcctc agcagtcaac
     1741 cgggtacgga cgatcgcgta acgtgggtga agtcggtgga tgaagccatc gcggcgtgtg
     1801 gtgacgtacc agaaatcatg gtgattggcg gcggtcgcgt ttatgaacag ttcttgccaa
     1861 aagcgcaaaa actgtatctg acgcatatcg acgcagaagt ggaaggcgac acccatttcc
     1921 cggattacga gccggatgac tgggaatcgg tattcagcga attccacgat gctgatgcgc
     1981 agaactctca cagctattgc tttgagattc tggagcggcg gggcagcggc catatgtaat
     2041 ctagagaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2101 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2161 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2221 tgtatcttat catgtctgga tccccgggta ccgagctcga attaattcct cttccgcttc
     2281 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2341 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2401 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2461 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2521 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2581 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2641 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2701 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2761 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2821 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2881 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2941 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3001 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3061 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3121 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3181 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3241 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3301 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3361 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3421 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3481 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3541 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3601 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3661 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3721 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3781 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3841 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3901 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3961 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4021 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4081 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4141 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4201 tgcaagcttg gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4261 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4321 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4381 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4441 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     4501 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcctcg
     4561 atcgaggggc tcgcatctct ccttcacgcg cccgctgccc tacctgaggc cgccatccac
     4621 gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg
     4681 tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc
     4741 tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct
     4801 ttgtttcgtt ttctgttctg cgccgttaca gatccaagct ctgaaaaacc agaaagttaa
     4861 ctggtaagtt tagtcttttt gtcttttatt tcaggtcccg gatccggtgg tggtgcaaat
     4921 caaagaactg ctcctcagtg gatgttgcct ttacttctag gcctgtacgg aagtgttact
     4981 tctg
//
