LOCUS       sequence-488309-        7858 bp    DNA     circular SYN 03-OCT-2025
DEFINITION  3rd gen lentiviral vector for knocking down p53 gene expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7858)
  AUTHORS   Godar S, Ince TA, Bell GW, Feldser D, Donaher JL, Bergh J, Liu A, 
            Miu K, Watnick RS, Reinhardt F, McAllister SS, Jacks T, Weinberg RA
  TITLE     Growth-inhibitory and tumor- suppressive functions of p53 depend on 
            its repression of CD44 expression.
  JOURNAL   Cell. 2008 Jul 11. 134(1):62-73.
  PUBMED    18614011
REFERENCE   2  (bases 1 to 7858)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 3, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2008 
            Jul 11. 134(1):62-73."
COMMENT     Sequence Label: shp53 pLKO.1 puro
FEATURES             Location/Qualifiers
     source          1..7858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  897..1664
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     promoter        join(1690..1695,1696..1713,1714..1720)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1728..1744
                     /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     1733..1755
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1752..1768
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1752..1768
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1787..1807
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1789..1807
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1835..2061
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2062..2242
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2289..2414
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2907..3140
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3325..3369
                     /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"
     promoter        3667..3915
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3667..3687
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3838..3857
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    4009..4126
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4175..4685
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     4604..4622
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             4707..5306
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4707..4726)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5203..5223
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5434..5667
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5739..5860
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5776..5795)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5830..5849
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5900..6035
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6055..6074)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6056..6074)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6084..6101)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6084..6100)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6093..6115)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6242..6697
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6329..6348)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6539..6560
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6723..6827
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6828..6896,6897..7688)
                     /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(7046..7065)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcggta
      901 atgactccaa cttattgata gtgttttatg ttcagataat gcccgatgac tttgtcatgc
      961 agctccaccg attttgagaa cgacagcgac ttccgtccca gccgtgccag gtgctgcctc
     1021 agattcaggt tatgccgctc aattcgctgc gtatatcgct tgctgattac gtgcagcttt
     1081 cccttcaggc gggattcata cagcggccag ccatccgtca tccatatcac cacgtcaaag
     1141 ggtgacagca ggctcataag acgccccagc gtcgccatag tgcgttcacc gaatacgtgc
     1201 gcaacaaccg tcttccggag actgtcatac gcgtaaaaca gccagcgctg gcgcgattta
     1261 gccccgacat agccccactg ttcgtccatt tccgcgcaga cgatgacgtc actgcccggc
     1321 tgtatgcgcg aggttaccga ctgcggcctg agttttttaa gtgacgtaaa atcgtgttga
     1381 ggccaacgcc cataatgcgg gctgttgccc ggcatccaac gccattcatg gccatatcaa
     1441 tgattttctg gtgcgtaccg ggttgagaag cggtgtaagt gaactgcagt tgccatgttt
     1501 tacggcagtg agagcagaga tagcgctgat gtccggcggt gcttttgccg ttacgcacca
     1561 ccccgtcagt agctgaacag gagggacagc tgatagaaac agaagccact ggagcacctc
     1621 aaaaacacca tcatacacta aatcagtaag ttggcagcat caccagctca ctcattaggc
     1681 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     1741 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca
     1801 ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt cttatgcaat actcttgtag
     1861 tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt
     1921 gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg
     1981 ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag
     2041 tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc
     2101 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     2161 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     2221 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac
     2281 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
     2341 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
     2401 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt
     2461 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc
     2521 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac
     2581 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata
     2641 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt
     2701 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg
     2761 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat
     2821 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg
     2881 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca
     2941 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct
     3001 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg
     3061 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac
     3121 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact
     3181 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg
     3241 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt
     3301 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg
     3361 gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa attattcata
     3421 atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat
     3481 agagttaggc agggatattc accattatcg tttcagaccc acctcccaac cccgagggga
     3541 cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt
     3601 cgattagtga acggatctcg acggtatcga tcacgagact agcctcgagc ggccgccccc
     3661 ttcaccgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     3721 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     3781 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     3841 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
     3901 aaaggacgaa acaccgactc cagtggtaat ctacttcaag agagtagatt accactggag
     3961 tctttttgaa ttctcgacct cgagacaaat ggcagtattc atccacaatt ttaaaagaaa
     4021 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     4081 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     4141 ggacagcaga gatccacttt ggccgcggct cgagggggtt ggggttgcgc cttttccaag
     4201 gcagccctgg gtttgcgcag ggacgcggct gctctgggcg tggttccggg aaacgcagcg
     4261 gcgccgaccc tgggtctcgc acattcttca cgtccgttcg cagcgtcacc cggatcttcg
     4321 ccgctaccct tgtgggcccc ccggcgacgc ttcctgctcc gcccctaagt cgggaaggtt
     4381 ccttgcggtt cgcggcgtgc cggacgtgac aaacggaagc cgcacgtctc actagtaccc
     4441 tcgcagacgg acagcgccag ggagcaatgg cagcgcgccg accgcgatgg gctgtggcca
     4501 atagcggctg ctcagcaggg cgcgccgaga gcagcggccg ggaaggggcg gtgcgggagg
     4561 cggggtgtgg ggcggtagtg tgggccctgt tcctgcccgc gcggtgttcc gcattctgca
     4621 agcctccgga gcgcacgtcg gcagtcggct ccctcgttga ccgaatcacc gacctctctc
     4681 cccaggggga tccaccggag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca
     4741 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     4801 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     4861 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     4921 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     4981 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     5041 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
     5101 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     5161 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     5221 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     5281 gcatgacccg caagcccggt gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa
     5341 ggagcgcacg accccatgca tcggtacctt taagaccaat gacttacaag gcagctgtag
     5401 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     5461 gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     5521 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     5581 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     5641 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     5701 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     5761 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5821 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     5881 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     5941 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     6001 attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat
     6061 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     6121 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     6181 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     6241 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     6301 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     6361 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     6421 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     6481 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     6541 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     6601 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     6661 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     6721 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6781 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     6841 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6901 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     6961 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     7021 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     7081 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     7141 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     7201 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     7261 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     7321 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     7381 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     7441 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7501 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     7561 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7621 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7681 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     7741 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     7801 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttc
//
