LOCUS       sequence-325292-        7380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Compact Cre expression vector with modified PPT designed to generate
            high-titer non-integrating lentivirus when packaged with PsPax2-D64V
            (Addgene #63586).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7380)
  AUTHORS   Dubrot J, Lane-Reticker SK, Kessler EA, Ayer A, Mishra G, Wolfe CH, 
            Zimmer MD, Du PP, Mahapatra A, Ockerman KM, Davis TGR, Kohnle IC, 
            Pope HW, Allen PM, Olander KE, Iracheta-Vellve A, Doench JG, Haining
            WN, Yates KB, Manguso RT
  TITLE     In vivo screens using a selective CRISPR antigen removal lentiviral 
            vector system reveal immune dependencies in renal cell carcinoma.
  JOURNAL   Immunity. 2021 Jan 20. pii: S1074-7613(21)00001-7. doi: 
            10.1016/j.immuni.2021.01.001.
  PUBMED    33497609
REFERENCE   2  (bases 1 to 7380)
  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: "Immunity. 
            2021 Jan 20. pii: S1074-7613(21)00001-7. doi: 
            10.1016/j.immuni.2021.01.001."
COMMENT     Sequence Label: pLX_EFS-Cre_ppt-del
FEATURES             Location/Qualifiers
     source          1..7380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    36..624
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(89..109)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(507..518)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             681..914
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    992..1113
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1029..1048)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1083..1102
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1153..1288
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1308..1327)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1309..1327)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1337..1354)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1337..1353)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1346..1368)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1495..1950
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1582..1601)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1792..1813
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1976..2080
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2081..2149,2150..2941)
                     /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     complement(2299..2318)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3112..3700
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3601..3620
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3854..3871
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3988..4009
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4024..4029,4030..4047,4048..4054)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4062..4078
                     /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     4067..4089
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4086..4102
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4086..4102
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4121..4141
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4123..4141
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4169..4395
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4396..4576
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4623..4748
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5241..5474
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5659..5703
                     /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    5890..6007
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6101..6312
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6331..6340
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 accggttaag tcgacaatca acgcgttaag tcgacaatca acctctggat tacaaaattt
       61 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
      121 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
      181 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
      241 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
      301 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
      361 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
      421 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
      481 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
      541 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
      601 tctccctttg ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt
      661 agatcttagc cactttttac tggaagggct aattcactcc caacgaagac aagatctgct
      721 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
      781 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
      841 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
      901 gaaaatctct agcagtacgt atagtagttc atgtcatctt attattcagt atttataact
      961 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     1021 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     1081 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     1141 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     1201 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     1261 aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt
     1321 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     1381 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     1441 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     1501 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     1561 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     1621 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     1681 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     1741 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     1801 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     1861 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     1921 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     1981 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2041 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     2101 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     2161 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     2221 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     2281 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     2341 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     2401 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     2461 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     2521 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     2581 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     2641 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     2701 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     2761 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     2821 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     2881 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     2941 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     3001 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3061 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3121 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3181 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3241 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3301 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3361 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3421 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3481 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3541 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3601 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3661 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3721 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     3781 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     3841 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     3901 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     3961 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     4021 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     4081 tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag
     4141 ggaacaaaag ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc
     4201 aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc
     4261 cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg
     4321 acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta
     4381 gctcgataca taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     4441 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     4501 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     4561 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
     4621 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
     4681 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
     4741 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
     4801 cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac
     4861 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac
     4921 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag
     4981 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca
     5041 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc
     5101 agacctggag gaggagatat gagggacaat tggagaagtg aattatataa atataaagta
     5161 gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga
     5221 gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc
     5281 actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata
     5341 gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc
     5401 acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag
     5461 gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg
     5521 ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg
     5581 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa
     5641 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt
     5701 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata
     5761 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt
     5821 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccatg
     5881 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     5941 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     6001 aaattttcgg gtttattaca gggacagcag agatccagtt tggttaatta agctagctag
     6061 gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg cacatcgccc
     6121 acagtccccg agaagttggg gggaggggtc ggcaattgat ccggtgccta gagaaggtgg
     6181 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
     6241 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
     6301 gccagaacac aggaccggtt ctagagcgct gccaccatgg ctaatctcct gaccgtgcat
     6361 cagaatctgc ctgccctgcc cgtcgacgca acaagcgatg aagtccgcaa gaatctcatg
     6421 gacatgttca gggacagaca ggccttttcc gagcacacct ggaagatgct gctgagcgtg
     6481 tgcaggtcct gggctgcttg gtgtaagctg aacaacagaa agtggttccc agctgagcca
     6541 gaggacgtgc gggattacct gctgtacctg caggcccgcg gactggctgt gaagacaatc
     6601 cagcagcacc tgggccagct gaacatgctg cacaggagaa gcggactgcc ccggcctagc
     6661 gactccaacg ccgtgagcct ggtcatgcgg cgcatcagga aggagaacgt ggatgccggc
     6721 gagagagcta agcaggccct ggctttcgag aggaccgact ttgatcaggt gagatctctg
     6781 atggagaaca gcgacaggtg ccaggatatc agaaacctgg cctttctggg aatcgcttac
     6841 aacaccctgc tgagaatcgc cgagatcgct cggatccgcg tgaaggacat ctctcggaca
     6901 gatggcggac gcatgctgat ccacatcggc aggaccaaga cactggtgtc caccgccggc
     6961 gtggagaagg ctctgtctct gggagtgaca aagctggtgg agagatggat ctctgtgagc
     7021 ggcgtggccg acgatcctaa caactacctg ttctgtaggg tgagaaagaa cggagtggcc
     7081 gctccatccg ctacctctca gctgagcaca cgggccctgg agggcatctt tgaggctacc
     7141 caccgcctga tctacggcgc caaggacgat tctggacagc ggtacctggc ttggtccgga
     7201 cactctgctc gcgtgggagc tgctcgggat atggcccgcg ctggcgtgag catcccagag
     7261 atcatgcagg ccggcggatg gacaaacgtg aacatcgtga tgaactacat tagaaatctg
     7321 gatagcgaaa ctggggcaat ggtgcggctg ctggaggatg gggactgata atgaactagt
//
