LOCUS       sequence-448452-        3515 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  packaging plasmid for lentivector production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3515)
  AUTHORS   Ziglari T, Calistri NL, Finan JM, Derrick DS, Nakayasu ES, Burnet 
            MC, Kyle JE, Hoare M, Heiser LM, Pucci F
  TITLE     Senescent Cell-Derived Extracellular Vesicles Inhibit Cancer 
            Recurrence by Coordinating Immune Surveillance.
  JOURNAL   Cancer Res. 2025 Mar 3;85(5):859-874. doi: 
            10.1158/0008-5472.CAN-24-0875.
  PUBMED    39804967
REFERENCE   2  (bases 1 to 3515)
  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"; doi: "10.1158/0008-5472";
            journalName: "Cancer Res"; date: "2025-03-3- 3"; volume: "85"; 
            issue: "5"; pages: "859-874"
COMMENT     Sequence Label: pKrev-CGI
FEATURES             Location/Qualifiers
     source          1..3515
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        34..413
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        414..617
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     568..588
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     662..681
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        662..680
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      766..775
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             772..1122
                     /codon_start=1
                     /gene="rev"
                     /product="human immunodeficiency virus 1 (HIV-1) Rev 
                     protein"
                     /label=Rev
                     /note="enhances nuclear export of unspliced and 
                     incompletely spliced viral mRNAs"
                     /translation="MAGRSGDSDEDLLKAVRLIKFLYQSNPPPNPEGTRQARRNRRRRW
                     RERQRQIHSISERILSTYLGRSAEPVPLQLPPLERLTLDCNEDCGTSGTQGVGSPQILV
                     ESPTILESGAKE"
     primer_bind     complement(1206..1223)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1206..1222)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1338..1355)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1344..1568
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             1741..2535
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1741..2535
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1795..1814)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2405..2424
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      2860..3448
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3349..3368
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat
       61 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      121 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      181 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      241 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      301 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
      361 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      421 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      481 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      541 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
      601 tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa
      661 ttaatacgac tcactatagg gagacccaag ctggctagcg tttaaactta agcttggtac
      721 cgagctcgga tccactagtc cagtgtggtg gaattcacgc gtgccgccac catggcagga
      781 agaagcggag acagcgacga agacctcctc aaggcagtca gactcatcaa gtttctctat
      841 caaagcaacc cacctcccaa tcccgagggg acccgacagg cccgaaggaa tagaagaaga
      901 aggtggagag agagacagag acagatccat tcgattagtg aacggatcct tagcacttat
      961 ctgggacgat ctgcggagcc tgtgcctctt cagctaccac cgcttgagag acttactctt
     1021 gattgtaacg aggattgtgg aacttctggg acgcaggggg tgggaagccc tcaaatattg
     1081 gtggaatctc ctacaatatt ggagtcagga gctaaagaat agtgctgtta gcttgctcaa
     1141 tgccacagct atagcagtag ctgaggggac agatagggtt atagaagtag tacaagaagc
     1201 ttggcactgg ccgtcgtttt acaacgtcgt gatctgagcc tgggagatcc actagtccag
     1261 tgtggtggaa ttctgcagat atccagcaca gtggcggccg ctcgagtcta gagggcccgt
     1321 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     1381 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     1441 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     1501 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     1561 ctctatggct tctactgggc ggttttatgg acagcaagcg aaccggaatt gccagctggg
     1621 gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca
     1681 aggatctgat ggcgcagggg atcaagctct gatcaagaga caggatgagg atcgtttcgc
     1741 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     1801 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     1861 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     1921 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     1981 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     2041 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     2101 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     2161 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     2221 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     2281 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     2341 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     2401 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     2461 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     2521 gacgagttct tctgaattat taacgcttac aatttcctga tgcggtattt tctccttacg
     2581 catctgtgcg gtatttcaca ccgcatcagg tggcactttt cggggaaatg tgcgcggaac
     2641 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2701 ctgataaatg cttcaataat agcacgtgct aaaacttcat ttttaattta aaaggatcta
     2761 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     2821 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2881 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2941 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3001 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3061 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3121 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3181 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3241 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3301 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3361 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3421 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3481 ggccttttgc tggccttttg ctcacatgtt cttgc
//
