LOCUS       sequence-8561-ex        6853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd gen lentiviral promoter-less Gateway destination vector, Zeo.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6853)
  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 6853)
  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 X1 Zeo DEST (668-1)
FEATURES             Location/Qualifiers
     source          1..6853
                     /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"
     protein_bind    3070..3194
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(3219..3224,3225..3242,3243..3249)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3303..3962
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3369..3388)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             4304..4609
                     /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     4488..4507
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4650..4774)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    4805..5393
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4858..4878)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5276..5287)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5444..5464)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5449..5779
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5630..5765
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5791..5838
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5857..6231
                     /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"
     LTR             6324..6557
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6629..6750
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6666..6685)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6720..6739
                     /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 cagatatcaa caagtttgta caaaaaagct gaacgagaaa cgtaaaatga tataaatatc
     3121 aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat
     3181 atccagtcac tatggcggcc gcattaggca ccccaggctt tacactttat gcttccggct
     3241 cgtataatgt gtggattttg agttaggatc cggcgagatt ttcaggagct aaggaagcta
     3301 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     3361 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     3421 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     3481 ttcacattct tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg
     3541 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     3601 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     3661 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     3721 agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg
     3781 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     3841 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtctgt gatggcttcc
     3901 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     3961 aaagatctgg atccggctta ctaaaagcca gataacagta tgcgtatttg cgcgctgatt
     4021 tttgcggtat aagaatatat actgatatgt atacccgaag tatgtcaaaa agaggtgtgc
     4081 tatgaagcag cgtattacag tgacagttga cagcgacagc tatcagttgc tcaaggcata
     4141 tatgatgtca atatctccgg tctggtaagc acaaccatgc agaatgaagc ccgtcgtctg
     4201 cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc ccggtttatt
     4261 gaaatgaacg gctcttttgc tgacgagaac agggactggt gaaatgcagt ttaaggttta
     4321 cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga
     4381 cacgcccggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt
     4441 ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac
     4501 cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc tcagccaccg
     4561 cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggctc
     4621 cgttatacac agccagtctg caggtcgacc atagtgactg gatatgttgt gttttacagt
     4681 attatgtagt ctgtttttta tgcaaaatct aatttaatat attgatattt atatcatttt
     4741 acgtttctcg ttcagctttc ttgtacaaag tggttgatat ccagcacagt ggcggccgct
     4801 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     4861 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     4921 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     4981 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     5041 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     5101 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     5161 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     5221 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     5281 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     5341 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat
     5401 tctgcagata tccggttagt aatgagtttg gaattaattc tgtggaatgt gtgtcagtta
     5461 gggtgtggaa agtccccagg ctccccaggc aggcagaagt atgcaaagca tgcatctcaa
     5521 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5581 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     5641 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5701 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     5761 aggcctaggc ttttgcaaaa agctccccct gttgacaatt aatcatcggc atagtatatc
     5821 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt
     5881 ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg
     5941 ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg
     6001 ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg
     6061 cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac
     6121 gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg
     6181 cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta
     6241 cgagatttaa atggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     6301 ctttttaaaa gaaaaggggg gactggaagg gctagctcac tcccaacgaa gacaagatct
     6361 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     6421 ctgcctaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     6481 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     6541 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     6601 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     6661 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6721 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     6781 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     6841 gaggccgagg ccg
//
