LOCUS       sequence-7003-ex        8633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lentiviral transfer vector. pPRIME-mating recipient 
            plasmid with CMV promoter controlling GFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8633)
  AUTHORS   Stegmeier F, Hu G, Rickles RJ, Hannon GJ, Elledge SJ
  TITLE     A lentiviral microRNA-based system for single-copy polymerase 
            II-regulated RNA interference in mammalian cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2005 Sep 13. 102(37):13212-7.
  PUBMED    16141338
REFERENCE   2  (bases 1 to 8633)
  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: "Proc Natl 
            Acad Sci U S A. 2005 Sep 13. 102(37):13212-7."
COMMENT     Sequence Label: pPRIME-CMV-GFP-recipient
FEATURES             Location/Qualifiers
     source          1..8633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    1023..1148
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1645..1878
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2063..2107
                     /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    2405..2522
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2659..2962
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2963..3166
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      3205..3214
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3211..3213,3214..3216,3217..3930)
                     /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(3211..3213,3214..3216,3217..3927)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3256..3277)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3517..3536)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3864..3885
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5370..5399
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    5522..6110
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5575..5595)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5993..6004)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6113..6129)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6639..6819
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6881..7469)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6961..6980)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7640..8431,8432..8500))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8263..8282
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8501..8605)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata gaccagatct gagcctggga gctctctggc
      841 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
      901 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
      961 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
     1021 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
     1081 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
     1141 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
     1201 cagggggaaa gaaaaaatat aaataaacat atagtatggg caagcaggga gctagaacga
     1261 ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag
     1321 ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca
     1381 accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag
     1441 atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgg ccgcgctgat
     1501 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     1561 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     1621 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     1681 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     1741 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     1801 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     1861 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     1921 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     1981 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     2041 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     2101 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     2161 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     2221 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     2281 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
     2341 attagtgaac ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc
     2401 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     2461 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     2521 ttcgggttta ttacagggac agcagagatc cagtttggtt agtaccgggc ccgctctaga
     2581 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
     2641 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     2701 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     2761 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     2821 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     2881 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     2941 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     3001 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     3061 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     3121 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
     3181 tcagatccgc tagcgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
     3241 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     3301 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     3361 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     3421 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     3481 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     3541 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     3601 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     3661 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     3721 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     3781 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     3841 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     3901 actctcggca tggacgagct gtacaagtag cggccgcaag ccttgttaag tgctcgcttc
     3961 ggcagcacat atactatgtt tgaatgaggc ttcagtactt tacagaatcg ttgcctgcac
     4021 atcttggaaa cacttgctgg gattacttct tcaggttaac ccaacagaag gccatgggtc
     4081 tagataggga taacagggta atggtaccgg atcccccaac ttccaggccc gccatagtga
     4141 tttgatttgc cagcgcatcg ctatcaatcg ccgggttcac ccattcgcgt aaccacagtt
     4201 cactgaattt cataatctat tcctgcctta tttaaactgt ttgaggaaac gcagatcgtt
     4261 ttcgaagaat gaacgcaggt cggtgacgcc gtaacgcaac atagtcagac gctccatccc
     4321 catcccgaag ccgaaaccag agtaaacttc cgggtcgatg ccaacgttac gcaacacgtt
     4381 cggatgcacc atcccgcagc ccagcacttc cagccattta ccgtttttac ccatgacgtc
     4441 cacttctgca gaaggttcgg taaacgggaa gtaggaagga cggaagcgaa tctgcaaatc
     4501 ttcctcaaag aagttacgca ggaagtcgtg cagcgtgcct ttcaggttgg taaagctgat
     4561 gttggtatca acaatcagac cttccatctg atggaacatc ggcgtgtgag tctggtcgta
     4621 gtcgttacga taaacacggc caggcgcgat gatacgaatc ggtggctgct gggctttcat
     4681 ggtgcggatc tgtacgccag aggtctgggt acgcagcagg cgggtagtgt caaaccagaa
     4741 agtgtcgtgg tcagcgcgcg ccgggtggtg accaggaatg ttcagagcat cgaagttatg
     4801 ataatcgtct tcgatttccg gcccggttgc cacggtaaag ccaagctcac cgaagaaact
     4861 ttcgatacgg tcgatggtac gggtaaccgg atgcagaccg ccgttttcaa tgcgacgacc
     4921 tggcagagag acatcaatcg tttccgccgc cagacgcgca ttcagtgcag cgctttccag
     4981 ttccgcttta cgcgcattca gcgcctgctg aacctgctct ttcgcttcgt tgataaccgc
     5041 accagctgcc ggacgctctt ctggcggcag ctcacgcagg gtcgtcatct gaagggttaa
     5101 gtgccctttt ttacccaaat attcgacgcg cacattatct aacgcggcaa catctgacgc
     5161 ctggctaatg gccgccttcg cactggcaac cagttctgcg agatgtgaca tggttttcct
     5221 cattgtgtca gtggtgacac tggttcgttg gacttagagc ctatcccatc aggctatttt
     5281 acttgccatt ttggtccccg ataccgtcga ccgcggatta ccctgttatc cctagatctt
     5341 tgggcggtgg ctccctgcca cgcggctccg aacagaagct gatctccgaa gaggatctga
     5401 catgtgttta aacctcgact taattaagtc gagggtcgac ggtatcgata agctcgcttc
     5461 acgagatcat gtttaagggt tccggttcca ctaggtacaa ttcgatatca agcttatcga
     5521 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     5581 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     5641 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     5701 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     5761 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     5821 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     5881 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     5941 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     6001 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     6061 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     6121 tcgacctcga tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta
     6181 ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca
     6241 cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     6301 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg
     6361 atctgtggat ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc
     6421 cagggatcag atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag
     6481 agaaggtaga agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc
     6541 atgggatgga tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat
     6601 ttcatcacat ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga
     6661 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     6721 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     6781 ccctcagacc cttttagtca gtgtggaaaa tctctagcag catgtgagca aaaggccagc
     6841 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6901 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6961 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7021 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     7081 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7141 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7201 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7261 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7321 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7381 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7441 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7501 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7561 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7621 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7681 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7741 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7801 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7861 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7921 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7981 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     8041 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     8101 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8161 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8221 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8281 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8341 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8401 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8461 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8521 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8581 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gac
//
