LOCUS       sequence-142555-        6368 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 6368)
  AUTHORS   Masutomi K, Yu EY, Khurts S, Ben-Porath I, Currier JL, Metz GB, 
            Brooks MW, Kaneko S, Murakami S, DeCaprio JA, Weinberg RA, Stewart 
            SA, Hahn WC
  TITLE     Telomerase maintains telomere structure in normal human cells.
  JOURNAL   Cell. 2003 Jul 25. 114(2):241-53.
  PUBMED    12887925
REFERENCE   2  (bases 1 to 6368)
  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: "Cell. 2003 
            Jul 25. 114(2):241-53."
COMMENT     Sequence Label: pMKO.1 puro p53 shRNA 1
FEATURES             Location/Qualifiers
     source          1..6368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        338..667
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      518..653
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             677..1276
                     /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(677..696)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1173..1193
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1472..1897
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(2000..2022)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        2163..2492
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2343..2478
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2514..2533
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2614..2631)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2785..3373)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2865..2884)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3544..4335,4336..4404))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4167..4186
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4405..4509)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4577..4595
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        4695..5074
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5075..5278
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5229..5249
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5279..5454
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    5517..5716
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5917..6333
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6263..6285
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6301..6317
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg gtgactccag tggtaatcta cttcaagaga gtagattaca ctggagtctt
      301 tttgaattcg ccagcacagt ggtcgaccct gtggaatgtg tgtcagttag ggtgtggaaa
      361 gtccccaggc ttcccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
      421 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
      481 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
      541 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
      601 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
      661 ttgcaaaaag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
      721 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
      781 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
      841 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
      901 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
      961 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     1021 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     1081 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     1141 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     1201 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     1261 caagcccggt gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg
     1321 accccatgca tcgattagtc caatttgtta aagacaggat atcagtggtc caggctctag
     1381 ttttgactca acaatatcac cagctgaagc ctatagagta cgagccatag ataaaataaa
     1441 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
     1501 agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga
     1561 agttcagatc aaggtcagga acagatggaa cagggtcgac cctagagaac catcagatgt
     1621 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     1681 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     1741 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
     1801 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     1861 tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga
     1921 gacccctgcc cagggaccac cgacccacca ccgggaggta agctggctgc ctcgcgcgtt
     1981 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     2041 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     2101 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtagatc cggctgtgga
     2161 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2221 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2281 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2341 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2401 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     2461 gaggcttttt tggaggccta ggcttttgca aaaagcttac tggcttaact atgcggcatc
     2521 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     2581 gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     2641 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2701 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2761 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2821 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2881 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2941 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3001 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3061 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3121 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3181 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     3241 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3301 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3361 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3421 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3481 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3541 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3601 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3661 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3721 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3781 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3841 caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3901 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3961 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4021 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4081 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4141 ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt
     4201 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4261 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4321 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4381 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4441 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4501 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     4561 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag aattagcttg
     4621 gccattgcat acgttgtatc catatcataa tatgtacatt tatattggct catgtccaac
     4681 attaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     4741 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     4801 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     4861 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     4921 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     4981 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     5041 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     5101 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     5161 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     5221 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctca
     5281 ataaaagagc ccacaacccc tcactcggcg cgccagtctt ccgatagact gcgtcgcccg
     5341 ggtacccgta ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg tctcgctgtt
     5401 ccttgggagg gtctcctctg agtgattgac tacccacgac gggggtcttt catttggggg
     5461 ctcgtccggg atttggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc
     5521 tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgtttga tgttatgcgc
     5581 ctgcgtctgt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
     5641 gagttctgaa cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt
     5701 tgtggcccga cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt
     5761 ctggtaggag acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt
     5821 tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc
     5881 tgtctgactg tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta
     5941 agtttgacct taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat
     6001 gtcaagaaga gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga
     6061 tggccgcgag acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt
     6121 tcacctggcc cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg
     6181 gcttttgacc cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt
     6241 cctccatccg ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc
     6301 ctttatccag ccctcactcc ttctctaggc gccggaattg aagatctgtc gacaaggtcg
     6361 ggcaggaa
//
