LOCUS       sequence-7005-ex        8719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lentiviral transfer vector. pPRIME-mating recipient 
            plasmid with CMV promoter controlling Neomycin expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8719)
  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 8719)
  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-Neo-recipient
FEATURES             Location/Qualifiers
     source          1..8719
                     /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"
     CDS             3212..4015
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3275..3294)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3885..3904
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             5456..5485
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    5608..6196
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5661..5681)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6079..6090)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6199..6215)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6725..6905
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6967..7555)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7047..7066)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7726..8517,8518..8586))
                     /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     8349..8368
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8587..8691)
                     /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 gggcctgcag gatgggatcg gccattgaac aagatggatt
     3241 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3301 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3361 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3421 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3481 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3541 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3601 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3661 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3721 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     3781 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3841 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3901 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     3961 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggc
     4021 cgcaagcctt gttaagtgct cgcttcggca gcacatatac tatgtttgaa tgaggcttca
     4081 gtactttaca gaatcgttgc ctgcacatct tggaaacact tgctgggatt acttcttcag
     4141 gttaacccaa cagaaggcca tgggtctaga tagggataac agggtaatgg taccggatcc
     4201 cccaacttcc aggcccgcca tagtgatttg atttgccagc gcatcgctat caatcgccgg
     4261 gttcacccat tcgcgtaacc acagttcact gaatttcata atctattcct gccttattta
     4321 aactgtttga ggaaacgcag atcgttttcg aagaatgaac gcaggtcggt gacgccgtaa
     4381 cgcaacatag tcagacgctc catccccatc ccgaagccga aaccagagta aacttccggg
     4441 tcgatgccaa cgttacgcaa cacgttcgga tgcaccatcc cgcagcccag cacttccagc
     4501 catttaccgt ttttacccat gacgtccact tctgcagaag gttcggtaaa cgggaagtag
     4561 gaaggacgga agcgaatctg caaatcttcc tcaaagaagt tacgcaggaa gtcgtgcagc
     4621 gtgcctttca ggttggtaaa gctgatgttg gtatcaacaa tcagaccttc catctgatgg
     4681 aacatcggcg tgtgagtctg gtcgtagtcg ttacgataaa cacggccagg cgcgatgata
     4741 cgaatcggtg gctgctgggc tttcatggtg cggatctgta cgccagaggt ctgggtacgc
     4801 agcaggcggg tagtgtcaaa ccagaaagtg tcgtggtcag cgcgcgccgg gtggtgacca
     4861 ggaatgttca gagcatcgaa gttatgataa tcgtcttcga tttccggccc ggttgccacg
     4921 gtaaagccaa gctcaccgaa gaaactttcg atacggtcga tggtacgggt aaccggatgc
     4981 agaccgccgt tttcaatgcg acgacctggc agagagacat caatcgtttc cgccgccaga
     5041 cgcgcattca gtgcagcgct ttccagttcc gctttacgcg cattcagcgc ctgctgaacc
     5101 tgctctttcg cttcgttgat aaccgcacca gctgccggac gctcttctgg cggcagctca
     5161 cgcagggtcg tcatctgaag ggttaagtgc ccttttttac ccaaatattc gacgcgcaca
     5221 ttatctaacg cggcaacatc tgacgcctgg ctaatggccg ccttcgcact ggcaaccagt
     5281 tctgcgagat gtgacatggt tttcctcatt gtgtcagtgg tgacactggt tcgttggact
     5341 tagagcctat cccatcaggc tattttactt gccattttgg tccccgatac cgtcgaccgc
     5401 ggattaccct gttatcccta gatctttggg cggtggctcc ctgccacgcg gctccgaaca
     5461 gaagctgatc tccgaagagg atctgacatg tgtttaaacc tcgacttaat taagtcgagg
     5521 gtcgacggta tcgataagct cgcttcacga gatcatgttt aagggttccg gttccactag
     5581 gtacaattcg atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga
     5641 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     5701 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     5761 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     5821 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     5881 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     5941 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     6001 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     6061 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     6121 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     6181 tgggccgcct ccccgcatcg ataccgtcga cctcgatcga gacctagaaa aacatggagc
     6241 aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga
     6301 ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa
     6361 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     6421 ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc
     6481 tgattggcag aactacacac cagggccagg gatcagatat ccactgacct ttggatggtg
     6541 ctacaagcta gtaccagttg agcaagagaa ggtagaagaa gccaatgaag gagagaacac
     6601 ccgcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtattaga
     6661 gtggaggttt gacagccgcc tagcatttca tcacatggcc cgagagctgc atccggactg
     6721 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     6781 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     6841 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     6901 tagcagcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6961 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7021 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7081 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7141 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7201 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7261 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7321 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7381 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     7441 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7501 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7561 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7621 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7681 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7741 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7801 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7861 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7921 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7981 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     8041 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     8101 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8161 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8221 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8281 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8341 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8401 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8461 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8521 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8581 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8641 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8701 ccgaaaagtg ccacctgac
//
