LOCUS       sequence-171211-        6474 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd gen lentiviral eGFP expression vector, CMV promoter, Zeo.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6474)
  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 6474)
  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 CMV GFP Zeo (637-7)
FEATURES             Location/Qualifiers
     source          1..6474
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      71..659
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     560..579
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     813..830
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    947..968
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(983..988,989..1006,1007..1013)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1021..1037
                     /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     1026..1048
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1045..1061
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1045..1061
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1080..1100
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1082..1100
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1128..1354
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1355..1535
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1582..1707
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2200..2433
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2618..2662
                     /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    2929..3045
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3068..3371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3372..3575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3526..3546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3572..3596
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      3665..3674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3671..3673,3674..3676,3677..4390)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3671..3673,3674..3676,3677..4387)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3716..3737)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3977..3996)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4324..4345
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4406..4994
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4459..4479)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4877..4888)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5045..5065)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5050..5379
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5230..5365
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5391..5438
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5457..5831
                     /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             5924..6157
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6229..6350
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6266..6285)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6320..6339
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
       61 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
      121 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      181 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
      241 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      301 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
      361 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
      421 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
      481 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
      541 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
      601 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
      661 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
      721 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
      781 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
      841 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
      901 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      961 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     1021 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
     1081 caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc
     1141 aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga
     1201 gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg
     1261 aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga
     1321 tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg
     1381 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     1441 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     1501 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
     1561 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
     1621 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
     1681 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
     1741 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
     1801 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
     1861 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
     1921 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
     1981 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
     2041 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     2101 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     2161 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     2221 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     2281 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     2341 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     2401 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     2461 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     2521 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     2581 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     2641 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     2701 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     2761 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     2821 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
     2881 agacagatcc attcgattag tgaacggatc tcgacggtat cggttaactt ttaaaagaaa
     2941 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     3001 acaaactaaa gaattacaaa aacaaattac aaaattcaaa attttatcga taagcttggg
     3061 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     3121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     3181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     3241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     3301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     3361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     3421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     3481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     3541 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct
     3601 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgactctag aggatccacc
     3661 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     3721 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     3781 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     3841 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     3901 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     3961 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     4021 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     4081 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     4141 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     4201 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     4261 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     4321 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     4381 gtacaagtaa agcggccgcg tcgacaatca acctctggat tacaaaattt gtgaaagatt
     4441 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     4501 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     4561 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     4621 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     4681 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     4741 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     4801 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     4861 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     4921 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     4981 ggccgcctcc ccgcctggaa ttctgcagat atccggttag taatgagttt ggaattaatt
     5041 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     5101 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     5161 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     5221 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     5281 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     5341 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctccccct gttgacaatt
     5401 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
     5461 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
     5521 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg
     5581 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
     5641 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
     5701 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
     5761 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
     5821 agcaggactg acacgtgcta cgagatttaa atggtacctt taagaccaat gacttacaag
     5881 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctagctcac
     5941 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     6001 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     6061 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     6121 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     6181 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
     6241 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     6301 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     6361 ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     6421 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccg
//
