LOCUS       sequence-71694-e        3766 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Contains the Gateway selection cassette (ccdB ChlorR) flanked by 
            AttR1 and AttR4 sites for the creation of Destination vectors .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3766)
  AUTHORS   Geiling B, Vandal G, Posner AR, de Bruyns A, Dutchak KL, Garnett S, 
            Dankort D
  TITLE     A modular lentiviral and retroviral construction system to rapidly 
            generate vectors for gene expression and gene knockdown in vitro and
            in vivo.
  JOURNAL   PLoS One. 2013 Oct 11;8(10):e76279. doi: 
            10.1371/journal.pone.0076279.
  PUBMED    24146852
REFERENCE   2  (bases 1 to 3766)
  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: "PLoS One.";
            date: "2013-10-11"; volume: "8(10):e76279. doi"; pages: " 
            10.1371/journal.pone.0076279"
COMMENT     Sequence Label: pBEG R1-ChlorR-R4   #BGV049
FEATURES             Location/Qualifiers
     source          1..3766
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    102..123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    178..194
                     /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)."
     primer_bind     183..205
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     202..218
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     202..218
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    249..373
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(404..409,410..427,428..434)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             488..1147
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(554..573)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1489..1794
                     /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
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1673..1692
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1856..1979)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2008..2027)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2009..2027)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2034..2051)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2034..2050)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2043..2065)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2257..3075)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSSRPRLNSNMDADLYGYKWARDNVGQSGATIY*LY
                     GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPG
                     KTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVD
                     ASDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVG
                     IADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2980..2999
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3067..3086
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3192..3736
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     3637..3656
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
       61 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
      121 cattaggcac cccaggcttt acactttatg cttccgcggc tcgtatgttg tgtggaattg
      181 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc actagcagat
      241 ctgctagcac aagtttgtac aaaaaagctg aacgagaaac gtaaaatgat ataaatatca
      301 atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa acacaacata
      361 tccagtcact atggccggcg cggccgcatt aggcacccca ggctttacac tttatgcttc
      421 cggctcgtat aatgtgtgga ttttgagtta ggatccggcg agattttcag gagctaagga
      481 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
      541 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
      601 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
      661 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa
      721 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
      781 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
      841 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
      901 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
      961 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac
     1021 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tctgtgatgg
     1081 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
     1141 ggcgtaatct agaggatccg gcttactaaa agccagataa cagtatgcgt atttgcgcgc
     1201 tgatttttgc ggtataagaa tatatactga tatgtatacc cgaagtatgt caaaaagagg
     1261 tgtgctatga agcagcgtat tacagtgaca gttgacagcg acagctatca gttgctcaag
     1321 gcatatatga tgtcaatatc tccggtctgg taagcacaac catgcagaat gaagcccgtc
     1381 gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg gatggctgag gtcgcccggt
     1441 ttattgaaat gaacggctct tttgctgacg agaacaggga ctggtgaaat gcagtttaag
     1501 gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt
     1561 attgacacgc ccgggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat
     1621 aaagtccccc gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg
     1681 accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc
     1741 caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca
     1801 ggctccctta tacacagcca gtctgcaggt cgacgatatc ggcgcgccac tagtccatag
     1861 tgactggata tgttgtgttt tacagtatta tgtagtctgt tttttatgca aaatctaatt
     1921 taatatattg atatttatat cattttacgt ttctcgttca acttttctat acaaagttgc
     1981 ccggggttaa cttcgatccc caattcgccc tatagtgagt cgtattacaa ttcactggcc
     2041 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     2101 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     2161 caacagttgc gcagcctgaa tggcgaatga gcttgcgccg tcccgtcaag tcagcgtaat
     2221 gctctgccag tgttacaacc aattaaccaa ttctgattag aaaaactcat cgagcatcaa
     2281 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     2341 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     2401 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     2461 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     2521 gttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     2581 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     2641 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     2701 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     2761 tttcccaggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     2821 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     2881 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     2941 atacaatcaa tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     3001 atataaatca gcatccatgt tggaatttaa tcgcggcctc gacgagcaag acgtttcccg
     3061 ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
     3121 tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acactcgaca
     3181 agatgatctt cttgagatcg ttttggtctg cgcgtaatct cttgctctga aaacgaaaaa
     3241 accgccttgc agggcggttt ttcgaaggtt ctctgagcta ccaactcttt gaaccgaggt
     3301 aactggcttg gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc ttaaccggcg
     3361 catgacttca agactaactc ctctaaatca attaccagtg gctgctgcca gtggtgcttt
     3421 tgcatgtctt tccgggttgg actcaagacg atagttaccg gataaggcgc agcggtcgga
     3481 ctgaacgggg ggttcgtgca tacagtccag cttggagcga actgcctacc cggaactgag
     3541 tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg aatgacaccg gtaaaccgaa
     3601 aggcaggaac aggagagcgc acgagggagc cgccagggga aacgcctggt atctttatag
     3661 tcctgtcggg tttcgccacc actgatttga gcgtcagatt tcgtgatgct tgtcaggggg
     3721 gcggagccta tggaaaaacg gctttgccgc ggccctctca cttccc
//
