LOCUS       sequence-440216-        8624 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lentiviral transfer vector. pPRIME-mating recipient 
            plasmid with CMV promoter controlling dsRed expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8624)
  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 8624)
  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-dsRed-recipient
FEATURES             Location/Qualifiers
     source          1..8624
                     /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"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1684..1917
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2102..2146
                     /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    2444..2561
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2698..3001
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3002..3205
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             3251..3928
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MDSSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(3381..3399)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(3431..3451)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     3649..3668
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3843..3866
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             5368..5397
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     5422..5438
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5513..6101
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5566..5586)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5984..5995)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6104..6120)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6630..6810
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6872..7460)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6952..6971)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7631..8422,8423..8491))
                     /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     8254..8273
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8492..8596)
                     /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 taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
     1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     2581 gcagagatcc agtttggtta gtaccgggcc cgctctagac gtattaccgc catgcattag
     2641 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     2701 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     2761 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     2821 ggtggagtat ttacggtaaa ctacccactt ggcagtacat caagtgtatc atatgccaag
     2881 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     2941 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     3001 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     3061 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     3121 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     3181 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg
     3241 ggcctgcagg atggactcct ccgaggacgt catcaaggag ttcatgcgct tcaaggtgcg
     3301 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     3361 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc
     3421 ctgggacatc ctgtcccccc agttccagta cggctccaag gtgtacgtga agcaccccgc
     3481 cgacatcccc gactacaaga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     3541 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggctc
     3601 cttcatctac aaggtgaagt tcatcggcgt gaacttcccc tccgacggcc ccgtaatgca
     3661 gaagaagact atgggctggg aggcctccac cgagcgcctg tacccccgcg acggcgtgct
     3721 gaagggcgag atccacaagg ccctgaagct gaaggacggc ggccactacc tggtggagtt
     3781 caagtccatc tacatggcca agaagcccgt gcagctgccc ggctactact acgtggactc
     3841 caagctggac atcacctccc acaacgagga ctacaccatc gtggagcagt acgagcgcgc
     3901 cgagggccgc caccacctgt tcctgtaggc ggccgcaagc cttgttaagt gctcgcttcg
     3961 gcagcacata tactatgttg aatgaggctt cagtacttta cagaatcgtt gcctgcacat
     4021 cttggaaaca cttgctggga ttacttcttc aggttaaccc aacagaaggc catgggtcta
     4081 gatagggata acagggtaat ggtaccggat cccccaactt ccaggcccgc catagtgatt
     4141 tgatttgcca gcgcatcgct atcaatcgcc gggttcaccc attcgcgtaa ccacagttca
     4201 ctgaatttca taatctattc ctgccttatt taaactgttt gaggaaacgc agatcgtttt
     4261 cgaagaatga acgcaggtcg gtgacgccgt aacgcaacat agtcagacgc tccatcccca
     4321 tcccgaagcc gaaaccagag taaacttccg ggtcgatgcc aacgttacgc aacacgttcg
     4381 gatgcaccat cccgcagccc agcacttcca gccatttacc gtttttaccc atgacgtcca
     4441 cttctgcaga aggttcggta aacgggaagt aggaaggacg gaagcgaatc tgcaaatctt
     4501 cctcaaagaa gttacgcagg aagtcgtgca gcgtgccttt caggttggta aagctgatgt
     4561 tggtatcaac aatcagacct tccatctgat ggaacatcgg cgtgtgagtc tggtcgtagt
     4621 cgttacgata aacacggcca ggcgcgatga tacgaatcgg tggctgctgg gctttcatgg
     4681 tgcggatctg tacgccagag gtctgggtac gcagcaggcg ggtagtgtca aaccagaaag
     4741 tgtcgtggtc agcgcgcgcc gggtggtgac caggaatgtt cagagcatcg aagttatgat
     4801 aatcgtcttc gatttccggc ccggttgcca cggtaaagcc aagctcaccg aagaaacttt
     4861 cgatacggtc gatggtacgg gtaaccggat gcagaccgcc gttttcaatg cgacgacctg
     4921 gcagagagac atcaatcgtt tccgccgcca gacgcgcatt cagtgcagcg ctttccagtt
     4981 ccgctttacg cgcattcagc gcctgctgaa cctgctcttt cgcttcgttg ataaccgcac
     5041 cagctgccgg acgctcttct ggcggcagct cacgcagggt cgtcatctga agggttaagt
     5101 gccctttttt acccaaatat tcgacgcgca cattatctaa cgcggcaaca tctgacgcct
     5161 ggctaatggc cgccttcgca ctggcaacca gttctgcgag atgtgacatg gttttcctca
     5221 ttgtgtcagt ggtgacactg gttcgttgga cttagagcct atcccatcag gctattttac
     5281 ttgccatttt ggtccccgat accgtcgacc gcggattacc ctgttatccc tagatctttg
     5341 ggcggtggct ccctgccacg cggctccgaa cagaagctga tctccgaaga ggatctgaca
     5401 tgtgtttaaa cttaattaag tcgaggtcga cggtatcgat aagctcgctt cacgagatca
     5461 tgtttaaggg ttccggttcc actaggtaca attcgatatc aagcttatcg ataatcaacc
     5521 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     5581 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     5641 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     5701 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     5761 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     5821 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     5881 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     5941 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     6001 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     6061 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacctcg
     6121 atcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg
     6181 attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg
     6241 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     6301 aggggggact ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga
     6361 tctaccacac acaaggctac ttccctgatt ggcagaacta cacaccaggg ccagggatca
     6421 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcaa gagaaggtag
     6481 aagaagccaa tgaaggagag aacacccgct tgttacaccc tgtgagcctg catgggatgg
     6541 atgacccgga gagagaagta ttagagtgga ggtttgacag ccgcctagca tttcatcaca
     6601 tggcccgaga gctgcatccg gactgtactg ggtctctctg gttagaccag atctgagcct
     6661 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6721 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     6781 ccttttagtc agtgtggaaa atctctagca gcatgtgagc aaaaggccag caaaaggcca
     6841 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6901 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6961 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     7021 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     7081 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7141 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7201 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7261 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     7321 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7381 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7441 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7501 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7561 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7621 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7681 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7741 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7801 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7861 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7921 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7981 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     8041 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     8101 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     8161 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8221 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8281 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8341 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8401 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8461 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8521 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8581 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgac
//
