LOCUS       sequence-7007-ex        8724 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lentiviral transfer vector. pPRIME-mating recipient 
            plasmid with Tet-responsive promoter (TREX) controlling GFP 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8724)
  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 8724)
  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-TREX-GFP-recipient
FEATURES             Location/Qualifiers
     source          1..8724
                     /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        2605..2984
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2985..3188
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    3190..3208
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3211..3229
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3239..3263
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      3296..3305
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3302..3304,3305..3307,3308..4021)
                     /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(3302..3304,3305..3307,3308..4018)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3347..3368)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3608..3627)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3955..3976
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5461..5490
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    5613..6201
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5666..5686)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6084..6095)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6204..6220)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6730..6910
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6972..7560)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7052..7071)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7731..8522,8523..8591))
                     /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     8354..8373
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8592..8696)
                     /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 atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa
     2641 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     2701 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2761 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2821 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2881 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2941 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     3001 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     3061 ttgacgtcaa tgggagtttg ttttggaacc aaaatcaacg ggactttcca aaatgtcgta
     3121 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     3181 gcagagctct ccctatcagt gatagagatc tccctatcag tgatagagat cgtcgacgag
     3241 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgttac cggtcgccac
     3301 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     3361 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     3421 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     3481 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     3541 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     3601 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     3661 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     3721 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     3781 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     3841 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     3901 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     3961 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     4021 gcggccgcaa gccttgttaa gtgctcgctt cggcagcaca tatactatgt ttgaatgagg
     4081 cttcagtact ttacagaatc gttgcctgca catcttggaa acacttgctg ggattacttc
     4141 ttcaggttaa cccaacagaa ggccatgggt ctagataggg ataacagggt aatggtaccg
     4201 gatcccccaa cttccaggcc cgccatagtg atttgatttg ccagcgcatc gctatcaatc
     4261 gccgggttca cccattcgcg taaccacagt tcactgaatt tcataatcta ttcctgcctt
     4321 atttaaactg tttgaggaaa cgcagatcgt tttcgaagaa tgaacgcagg tcggtgacgc
     4381 cgtaacgcaa catagtcaga cgctccatcc ccatcccgaa gccgaaacca gagtaaactt
     4441 ccgggtcgat gccaacgtta cgcaacacgt tcggatgcac catcccgcag cccagcactt
     4501 ccagccattt accgttttta cccatgacgt ccacttctgc agaaggttcg gtaaacggga
     4561 agtaggaagg acggaagcga atctgcaaat cttcctcaaa gaagttacgc aggaagtcgt
     4621 gcagcgtgcc tttcaggttg gtaaagctga tgttggtatc aacaatcaga ccttccatct
     4681 gatggaacat cggcgtgtga gtctggtcgt agtcgttacg ataaacacgg ccaggcgcga
     4741 tgatacgaat cggtggctgc tgggctttca tggtgcggat ctgtacgcca gaggtctggg
     4801 tacgcagcag gcgggtagtg tcaaaccaga aagtgtcgtg gtcagcgcgc gccgggtggt
     4861 gaccaggaat gttcagagca tcgaagttat gataatcgtc ttcgatttcc ggcccggttg
     4921 ccacggtaaa gccaagctca ccgaagaaac tttcgatacg gtcgatggta cgggtaaccg
     4981 gatgcagacc gccgttttca atgcgacgac ctggcagaga gacatcaatc gtttccgccg
     5041 ccagacgcgc attcagtgca gcgctttcca gttccgcttt acgcgcattc agcgcctgct
     5101 gaacctgctc tttcgcttcg ttgataaccg caccagctgc cggacgctct tctggcggca
     5161 gctcacgcag ggtcgtcatc tgaagggtta agtgcccttt tttacccaaa tattcgacgc
     5221 gcacattatc taacgcggca acatctgacg cctggctaat ggccgccttc gcactggcaa
     5281 ccagttctgc gagatgtgac atggttttcc tcattgtgtc agtggtgaca ctggttcgtt
     5341 ggacttagag cctatcccat caggctattt tacttgccat tttggtcccc gataccgtcg
     5401 accgcggatt accctgttat ccctagatct ttgggcggtg gctccctgcc acgcggctcc
     5461 gaacagaagc tgatctccga agaggatctg acatgtgttt aaacctcgac ttaattaagt
     5521 cgagggtcga cggtatcgat aagctcgctt cacgagatca tgtttaaggg ttccggttcc
     5581 actaggtaca attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
     5641 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     5701 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     5761 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     5821 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     5881 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     5941 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     6001 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     6061 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     6121 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     6181 ccctttgggc cgcctccccg catcgatacc gtcgacctcg atcgagacct agaaaaacat
     6241 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca
     6301 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact
     6361 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     6421 attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac
     6481 ttccctgatt ggcagaacta cacaccaggg ccagggatca gatatccact gacctttgga
     6541 tggtgctaca agctagtacc agttgagcaa gagaaggtag aagaagccaa tgaaggagag
     6601 aacacccgct tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagta
     6661 ttagagtgga ggtttgacag ccgcctagca tttcatcaca tggcccgaga gctgcatccg
     6721 gactgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     6781 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     6841 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     6901 atctctagca gcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6961 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     7021 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     7081 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7141 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     7201 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7261 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7321 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7381 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     7441 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7501 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7561 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7621 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7681 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7741 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7801 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7861 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7921 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7981 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     8041 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     8101 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     8161 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8221 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     8281 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8341 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8401 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8461 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8521 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8581 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8641 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8701 tttccccgaa aagtgccacc tgac
//
