LOCUS       sequence-254695-        7083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7083)
  AUTHORS   Kim JS, Lee C, Bonifant CL, Ressom H, Waldman T
  TITLE     Activation of p53-dependent growth suppression in human cells by 
            mutations in PTEN or PIK3CA.
  JOURNAL   Mol Cell Biol. 2007 Jan . 27(2):662-77.
  PUBMED    17060456
REFERENCE   2  (bases 1 to 7083)
  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: "Mol Cell 
            Biol. 2007 Jan . 27(2):662-77."
COMMENT     Sequence Label: pLKO-p53-shRNA-941
FEATURES             Location/Qualifiers
     source          1..7083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..234
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     CDS             complement(362..961)
                     /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(445..465)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     942..961
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(983..1493)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1046..1064)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     misc_feature    1542..1659
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        complement(1755..2003)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(1813..1832)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1983..2003)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     CDS             complement(2301..2345)
                     /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    2530..2763
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    3256..3381
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             3428..3608
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(3609..3835)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(3863..3883)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3863..3881)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3902..3918)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3902..3918)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3915..3937)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3926..3942
                     /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)."
     promoter        complement(join(3950..3956,3957..3974,3975..3980))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3995..4016
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4133..4150)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4304..4892)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4384..4403)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5063..5854,5855..5923))
                     /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     5686..5705
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5924..6028)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6054..6509)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6191..6212)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6403..6422
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6636..6658
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6650..6667
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6651..6667
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6677..6696
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6677..6695
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      6716..6851
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6891..7012
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6902..6921)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6956..6975
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta ccagagtcac
       61 acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga ggcttaagca
      121 gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac cagagagacc
      181 cagtacaagc aaaaagcaga tcttgtcttc gttgggagtg aattagccct tccagtcccc
      241 ccttttcttt taaaaagtgg ctaagatcta cagctgcctt gtaagtcatt ggtcttaaag
      301 gtaccgatgc atggggtcgt gcgctccttt cggtcgggcg ctgcgggtcg tggggcgggc
      361 gtcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc acctcgacgt
      421 cggcggtgac ggtgaagccg agccgctcgt agaaggggag gttgcggggc gcggaggtct
      481 ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac tccggggagc acgacggcgc
      541 tgcccagacc cttgccctgg tggtcgggcg agacgccgac ggtggccagg aaccacgcgg
      601 gctccttggg ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg gccagccggg
      661 aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa caccgccccc gcttcgacgc
      721 tctccggcgt ggtccagacc gccaccgcgg cgccgtcgtc cgcgacccac accttgccga
      781 tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc tcgatgtggc
      841 ggtccggatc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg gcgagggtgc
      901 gtacggccct ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg tactcggtca
      961 tggtaagctc cggtggatcc ccctggggag agaggtcggt gattcggtca acgagggagc
     1021 cgactgccga cgtgcgctcc ggaggcttgc agaatgcgga acaccgcgcg ggcaggaaca
     1081 gggcccacac taccgcccca caccccgcct cccgcaccgc cccttcccgg ccgctgctct
     1141 cggcgcgccc tgctgagcag ccgctattgg ccacagccca tcgcggtcgg cgcgctgcca
     1201 ttgctccctg gcgctgtccg tctgcgaggg tactagtgag acgtgcggct tccgtttgtc
     1261 acgtccggca cgccgcgaac cgcaaggaac cttcccgact taggggcgga gcaggaagcg
     1321 tcgccggggg gcccacaagg gtagcggcga agatccgggt gacgctgcga acggacgtga
     1381 agaatgtgcg agacccaggg tcggcgccgc tgcgtttccc ggaaccacgc ccagagcagc
     1441 cgcgtccctg cgcaaaccca gggctgcctt ggaaaaggcg caaccccaac cccctcgagc
     1501 cgcggccaaa gtggatctct gctgtccctg taataaaccc gaaaattttg aatttttgta
     1561 atttgttttt gtaattcttt agtttgtatg tctgttgcta ttatgtctac tattctttcc
     1621 cctgcactgt accccccaat cccccctttt cttttaaaat tgtggatgaa tactgccatt
     1681 tgtctcgagg tcgagaatta aaaacaccat ccactacaac tacatctcga gatgtagttg
     1741 tagtggatgg tgccggtgtt tcgtcctttc cacaagatat ataaagccaa gaaatcgaaa
     1801 tactttcaag ttacggtaag catatgatag tccattttaa aacataattt taaaactgca
     1861 aactacccaa gaaattatta ctttctacgt cacgtatttt gtactaatat ctttgtgttt
     1921 acagtcaaat taattccaat tatctctcta acagccttgt atcgtatatg caaatatgaa
     1981 ggaatcatgg gaaataggcc ctcggtgaag ggggcggccg ctcgaggcta gtctcgtgat
     2041 cgataccgtc gagatccgtt cactaatcga atggatctgt ctctgtctct ctctccacct
     2101 tcttcttcta ttccttcggg cctgtcgggt cccctcgggg ttgggaggtg ggtctgaaac
     2161 gataatggtg aatatccctg cctaactcta ttcactatag aaagtacagc aaaaactatt
     2221 cttaaaccta ccaagcctcc tactatcatt atgaataatt ttatatacca cagccaattt
     2281 gttatgttaa accaattcca caaacttgcc catttatcta attccaataa ttcttgttca
     2341 ttcttttctt gctggttttg cgattcttca attaaggagt gtattaagct tgtgtaattg
     2401 ttaatttctc tgtcccactc catccaggtc gtgtgattcc aaatctgttc cagagattta
     2461 ttactccaac tagcattcca aggcacagca gtggtgcaaa tgagttttcc agagcaaccc
     2521 caaatcccca ggagctgttg atcctttagg tatctttcca cagccaggat tcttgcctgg
     2581 agctgcttga tgccccagac tgtgagttgc aacagatgct gttgcgcctc aatagccctc
     2641 agcaaattgt tctgctgctg cactatacca gacaataatt gtctggcctg taccgtcagc
     2701 gtcattgacg ctgcgcccat agtgcttcct gctgctccca agaacccaag gaacaaagct
     2761 cctattccca ctgctctttt ttctctctgc accactcttc tctttgcctt ggtgggtgct
     2821 actcctaatg gttcaatttt tactacttta tatttatata attcacttct ccaattgtcc
     2881 ctcatatctc ctcctccagg tctgaagatc agcggccgct tgctgtgcgg tggtcttact
     2941 tttgttttgc tcttcctcta tcttgtctaa agcttccttg gtgtctttta tctctatcct
     3001 ttgatgcaca caatagaggg ttgctactgt attatataat gatctaagtt cttctgatcc
     3061 tgtctgaagg gatggttgta gctgtcccag tatttgtcta cagccttctg atgtttctaa
     3121 caggccagga ttaactgcga atcgttctag ctccctgctt gcccatacta tatgttttaa
     3181 tttatatttt ttctttcccc ctggccttaa ccgaattttt tcccatcgcg atctaattct
     3241 cccccgctta atactgacgc tctcgcaccc atctctctcc ttctagcctc cgctagtcaa
     3301 aatttttggc gtactcacca gtcgccgccc ctcgcctctt gccgtgcgcg cttcagcaag
     3361 ccgagtcctg cgtcgagaga gctcctctgg tttccctttc gctttcaagt ccctgttcgg
     3421 gcgccactgc tagagatttt ccacactgac taaaagggtc tgagggatct ctagttacca
     3481 gagtcacaca acagacgggc acacactact tgaagcactc aaggcaagct ttattgaggc
     3541 ttaagcagtg ggttccctag ttagccagag agctcccagg ctcagatctg gtctaaccag
     3601 agagacccgt ttatgtatcg agctaggcac ttaaatacaa tatctctgca atgcggcaat
     3661 tcagtggttc gtccaatcca tgtcagaccc gtctgttgcc ttcctaataa ggcacgatcg
     3721 taccacctta cttccaccaa tcggcatgca cggtgctttt tctctccttg taaggcatgt
     3781 tgctaactca tcgttaccat gttgcaagac tacaagagta ttgcataaga ctacattaag
     3841 cttgcagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     3901 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3961 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4021 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4081 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4141 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4201 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4261 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4321 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4381 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4441 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4501 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4561 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4621 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4681 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4741 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4801 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4861 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4921 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4981 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5041 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5101 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5161 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5221 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5281 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5341 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5401 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5461 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5521 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5581 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5641 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5701 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5761 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5821 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5881 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5941 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6001 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
     6061 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     6121 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     6181 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     6241 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     6301 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     6361 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     6421 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     6481 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
     6541 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     6601 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     6661 ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtacgt ccctaggcct
     6721 ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct
     6781 gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt
     6841 aggggcggga tgggcggagt taggggcggg atagctagag ccagacatga taagatacat
     6901 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     6961 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttcctctcac
     7021 tctctgatat tcatttcttt gcaagttata aatactgaat aataagatga catgaactac
     7081 tac
//
