LOCUS       sequence-9917-ex        6909 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd gen lentiviral Gateway destination vector, RNAi, 3'LTR 
            insertion, Zeo.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6909)
  AUTHORS   Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, 
            Campisi J, Yaswen P, Cooper PK, Kaufman PD.
  TITLE     A versatile viral system for expression and depletion of proteins in
            mammalian cells.
  JOURNAL   PLoS One. 2009 Aug 6;4(8):e6529.
  PUBMED    19657394
REFERENCE   2  (bases 1 to 6909)
  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: "2009-08-6"; volume: "4(8)"; pages: "e6529"
COMMENT     Sequence Label: pLenti X2 Zeo DEST (749-3)
FEATURES             Location/Qualifiers
     source          1..6909
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      72..660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     561..580
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     814..831
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    948..969
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(984..989,990..1007,1008..1014)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1022..1038
                     /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     1027..1049
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1046..1062
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1046..1062
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1081..1101
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1083..1101
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     LTR             1357..1537
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1581..1706
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2199..2432
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2616..2660
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2921..3038
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3092..3680
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3145..3165)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3563..3574)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3731..3751)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3736..4066
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3917..4052
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4078..4125
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4144..4518
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     protein_bind    4654..4778
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(4819..5124)
                     /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     complement(4921..4940)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(5466..6125)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6040..6059
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(6179..6185,6186..6203,6204..6209))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(6234..6358)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     LTR             6433..6613
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    6685..6806
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6722..6741)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6776..6795
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
       61 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
      121 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
      181 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
      241 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
      301 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
      361 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
      421 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
      481 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
      541 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
      601 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
      661 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      721 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      781 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      841 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      901 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      961 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     1021 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
     1081 gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg tagtcttatg
     1141 caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag
     1201 agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag
     1261 gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag
     1321 atattgtatt taagtgccta gctcgataca ataaacgggt ctctctggtt agaccagatc
     1381 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     1441 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     1501 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacctga
     1561 aagcgaaagg gaaaccagag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
     1621 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
     1681 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
     1741 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
     1801 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
     1861 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1921 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1981 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     2041 aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa
     2101 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag
     2161 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc
     2221 ttgggagcag caggaagcac tatgggcgca gcctcaatga cgctgacggt acaggccaga
     2281 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
     2341 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
     2401 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
     2461 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
     2521 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
     2581 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat
     2641 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa
     2701 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt
     2761 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa
     2821 ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag
     2881 acagatccat tcgattagtg aacggatctc gacggttaac ttttaaaaga aaagggggga
     2941 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     3001 aagaattaca aaaacaaatt acaaaaattc aaaattttat tccagtgtgg tggaattctg
     3061 cagatatcca gcacagtggc ggccgctcga caatcaacct ctggattaca aaatttgtga
     3121 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     3181 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     3241 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     3301 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     3361 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     3421 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     3481 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     3541 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3601 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3661 cctttgggcc gcctccccgc ctggaattct gcagatatcc ggttagtaat gagtttggaa
     3721 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccaggcagg
     3781 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3841 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3901 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3961 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     4021 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tccccctgtt
     4081 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa
     4141 accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg
     4201 gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc
     4261 ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg
     4321 gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg
     4381 gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag
     4441 cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg
     4501 gccgaggagc aggactgaca cgtgctacga gatttaaatg gtacctttaa gaccaatgac
     4561 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     4621 agactcgagc ggccgccact gtgctggata tcaaccactt tgtacaagaa agctgaacga
     4681 gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact
     4741 acataatact gtaaaacaca acatatccag tcactatggt cgacctgcag actggctgtg
     4801 tataacggag cctgacattt atattcccca gaacatcagg ttaatggcgt ttttgatgtc
     4861 attttcgcgg tggctgagat cagccacttc ttccccgata acggagaccg gcacactggc
     4921 catatcggtg gtcatcatgc gccagctttc atccccgata tgcaccaccg ggtaaagttc
     4981 acgggagact ttatctgaca gcagacgtgc actggccagg gggatcacca tccgtcgccc
     5041 gggcgtgtca ataatatcac tctgtacatc cacaaacaga cgataacggc tctctctttt
     5101 ataggtgtaa accttaaact gcatttcacc agtccctgtt ctcgtcagca aaagagccgt
     5161 tcatttcaat aaaccgggcg acctcagcca tcccttcctg attttccgct ttccagcgtt
     5221 cggcacgcag acgacgggct tcattctgca tggttgtgct taccagaccg gagatattga
     5281 catcatatat gccttgagca actgatagct gtcgctgtca actgtcactg taatacgctg
     5341 cttcatagca cacctctttt tgacatactt cgggtataca tatcagtata tattcttata
     5401 ccgcaaaaat cagcgcgcaa atacgcatac tgttatctgg cttttagtaa gccggatcca
     5461 gatctttacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg
     5521 ccgacatgga agccatcaca gacggcatga tgaacctgaa tcgccagcgg catcagcacc
     5581 ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata
     5641 ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac
     5701 atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct
     5761 tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa
     5821 aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc
     5881 agctcaccgt ctttcattgc catacggaat tccggatgag cattcatcag gcgggcaaga
     5941 atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc
     6001 gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca
     6061 aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc
     6121 tccattttag cttccttagc tcctgaaaat ctcgccggat cctaactcaa aatccacaca
     6181 ttatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgcggc cgccatagtg
     6241 actggatatg ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta
     6301 atatattgat atttatatca ttttacgttt ctcgttcagc ttttttgtac aaacttgttg
     6361 atatctgcag aattccacca cactggacta gctcactccc aacgaagaca agatctgctt
     6421 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctgc
     6481 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     6541 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     6601 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     6661 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
     6721 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     6781 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc
     6841 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     6901 ccgaggccg
//
