LOCUS       sequence-71715-e        3703 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Supplies the selection marker (eCFP) and IRES with flanked by AttR2 
            and AttL3 sites for Gateway cloning recombination reaction. (This 
            corresponds to a module 3 plasmid)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3703)
  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 3703)
  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 R2-eCFP-L3   #BGV202
FEATURES             Location/Qualifiers
     source          1..3703
                     /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    complement(249..373)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    join(425..999,1000..1002,1003..1011)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(592..609)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     819..838
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1042..1044,1045..1047,1048..1761)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1087..1108)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1348..1367)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1695..1716
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1821..1916)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1945..1964)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1946..1964)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1971..1988)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1971..1987)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1980..2002)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2194..3012)
                     /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     2917..2936
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3004..3023
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3129..3673
                     /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     3574..3593
                     /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 ctgccggcca tagtgactgg atatgttgtg ttttacagta ttatgtagtc tgttttttat
      301 gcaaaatcta atttaatata ttgatattta tatcatttta cgtttctcgt tcagctttct
      361 tgtacaaagt ggtgctagcg cggccgcccc cttcaccttg gatccgccgg cgaattccgc
      421 cccgcccctc tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc
      481 ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg
      541 cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca
      601 aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa
      661 gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt
      721 gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt
      781 gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca
      841 acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc
      901 ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc
      961 acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaac catggccaga
     1021 tccgctgagg agacatctag aatggtgagc aagggcgagg agctgttcac cggggtggtg
     1081 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1141 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1201 ctgcccgtgc cctggcccac cctcgtgacc accctgacct ggggcgtgca gtgcttcagc
     1261 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1321 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1381 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1441 gacggcaaca tcctggggca caagctggag tacaactaca tcagccacaa cgtctatatc
     1501 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     1561 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1621 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     1681 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1741 atggacgagc tgtacaagta atctagactt tccctcagcg gatctcctag gctcgagtta
     1801 agggtgggcg cgcccccggg caactttatt atacaaagtt ggcattataa aaaagcattg
     1861 ctcatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttacta
     1921 gtgttaactt cgatccccaa ttcgccctat agtgagtcgt attacaattc actggccgtc
     1981 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     2041 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     2101 cagttgcgca gcctgaatgg cgaatgagct tgcgccgtcc cgtcaagtca gcgtaatgct
     2161 ctgccagtgt tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg
     2221 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     2281 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     2341 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     2401 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaaggtt
     2461 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     2521 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     2581 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     2641 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     2701 cccagggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     2761 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     2821 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     2881 caatcaatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     2941 taaatcagca tccatgttgg aatttaatcg cggcctcgac gagcaagacg tttcccgttg
     3001 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
     3061 tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca ctcgacaaga
     3121 tgatcttctt gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc
     3181 gccttgcagg gcggtttttc gaaggttctc tgagctacca actctttgaa ccgaggtaac
     3241 tggcttggag gagcgcagtc accaaaactt gtcctttcag tttagcctta accggcgcat
     3301 gacttcaaga ctaactcctc taaatcaatt accagtggct gctgccagtg gtgcttttgc
     3361 atgtctttcc gggttggact caagacgata gttaccggat aaggcgcagc ggtcggactg
     3421 aacggggggt tcgtgcatac agtccagctt ggagcgaact gcctacccgg aactgagtgt
     3481 caggcgtgga atgagacaaa cgcggccata acagcggaat gacaccggta aaccgaaagg
     3541 caggaacagg agagcgcacg agggagccgc caggggaaac gcctggtatc tttatagtcc
     3601 tgtcgggttt cgccaccact gatttgagcg tcagatttcg tgatgcttgt caggggggcg
     3661 gagcctatgg aaaaacggct ttgccgcggc cctctcactt ccc
//
