LOCUS       sequence-382461-        6281 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Gateway destination vector for mammalian expression of Virotrap bait
            constructs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6281)
  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 6281)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-GW
FEATURES             Location/Qualifiers
     source          1..6281
                     /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"
     protein_bind    1620..1744
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1769..1774,1775..1792,1793..1799)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1853..2512
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1919..1938)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2854..3159
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3038..3057
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3200..3324)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    3405..3539
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3442..3461)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3496..3515
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3578..3595)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3749..4337)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3829..3848)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4508..5299,5300..5368))
                     /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     5131..5150
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5369..5473)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5513..5533)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(5518..5847,5848..5862,5863..6131)
                     /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      5698..5833
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     LTR             5863..6131
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     intron          6161..6257
                     /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 caagtttgta caaaaaagct gaacgagaaa cgtaaaatga tataaatatc aatatattaa
     1681 attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat atccagtcac
     1741 tatggcggcc gcattaggca ccccaggctt tacactttat gcttccggct cgtataatgt
     1801 gtggattttg agttaggatc cgtcgagatt ttcaggagct aaggaagcta aaatggagaa
     1861 aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga
     1921 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc
     1981 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct
     2041 tgcccgcctg atgaatgctc atccggagtt ccgtatggca atgaaagacg gtgagctggt
     2101 gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc
     2161 atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga
     2221 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt
     2281 tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat
     2341 ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt
     2401 gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag
     2461 aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt aaagatctgg
     2521 atccggctta ctaaaagcca gataacagta tgcgtatttg cgcgctgatt tttgcggtat
     2581 aagaatatat actgatatgt atacccgaag tatgtcaaaa agaggtatgc tatgaagcag
     2641 cgtattacag tgacagttga cagcgacagc tatcagttgc tcaaggcata tatgatgtca
     2701 atatctccgg tctggtaagc acaaccatgc agaatgaagc ccgtcgtctg cgtgccgaac
     2761 gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc ccggtttatt gaaatgaacg
     2821 gctcttttgc tgacgagaac aggggctggt gaaatgcagt ttaaggttta cacctataaa
     2881 agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga cacgcccggg
     2941 cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt ctcccgtgaa
     3001 ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac cgatatggcc
     3061 agtgtgccgg tctccgttat cggggaagaa gtggctgatc tcagccaccg cgaaaatgac
     3121 atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggctc ccttatacac
     3181 agccagtctg caggtcgacc atagtgactg gatatgttgt gttttacagt attatgtagt
     3241 ctgtttttta tgcaaaatct aatttaatat attgatattt atatcatttt acgtttctcg
     3301 ttcagctttc ttgtacaaag tggtttaact cgagagtcta gagaaaaaac ctcccacacc
     3361 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3421 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3481 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc
     3541 ccgggtaccg agctcgaatt aattcctctt ccgcttcctc gctcactgac tcgctgcgct
     3601 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3661 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3721 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3781 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3841 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3901 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3961 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4021 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4081 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4141 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4201 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4261 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4321 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4381 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4441 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4501 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4561 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4621 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4681 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4741 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4801 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4861 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4921 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4981 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5041 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5101 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5161 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5221 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5281 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5341 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5401 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5461 taggggttcc gcgcacattt ccccgaaaag tgccacctgc aagcttggct gtggaatgtg
     5521 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5581 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     5641 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     5701 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5761 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     5821 ttttttggag gcctaggctt ttgcaaaaag ctcctcgatc gaggggctcg catctctcct
     5881 tcacgcgccc gctgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg
     5941 cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg
     6001 tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc
     6061 acgctttgcc tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc
     6121 cgttacagat ccaagctctg aaaaaccaga aagttaactg gtaagtttag tctttttgtc
     6181 ttttatttca ggtcccggat ccggtggtgg tgcaaatcaa agaactgctc ctcagtggat
     6241 gttgccttta cttctaggcc tgtacggaag tgttacttct g
//
