LOCUS       Exported                6370 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pMKO.1 puro p53 shRNA 2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6370)
  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 6370)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6370
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        19..259
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     19..39
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     190..209
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        358..687
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      538..673
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             697..1296
                     /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(697..716)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1193..1213
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1492..1917
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(2020..2042)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        2183..2512
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2363..2498
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2534..2553
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2634..2651)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2805..3393)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2885..2904)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3564..4424)
                     /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     4187..4206
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4425..4529)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4597..4615
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        4715..5094
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5095..5298
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5249..5269
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5299..5474
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    5537..5736
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5937..6353
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6283..6305
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6321..6337
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 gacaaggtcg ggcaggaaga gggcctattt cccatgattc cttcatattt gcatatacga
       61 tacaaggctg ttagagagat aattagaatt aatttgactg taaacacaaa gatattagta
      121 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt
      181 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta
      241 tatatcttgt ggaaaggacg aaacaccggt gactccagtg gtaatctact gctcgagcag
      301 tagattacca ctggagtctt tttgaattcg ccagcacagt ggtcgaccct gtggaatgtg
      361 tgtcagttag ggtgtggaaa gtccccaggc ttcccagcag gcagaagtat gcaaagcatg
      421 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
      481 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
      541 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
      601 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
      661 ttttttggag gcctaggctt ttgcaaaaag cttaccatga ccgagtacaa gcccacggtg
      721 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
      781 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
      841 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
      901 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
      961 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     1021 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     1081 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     1141 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     1201 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     1261 cgcacctggt gcatgacccg caagcccggt gcctgacgcc cgccccacga cccgcagcgc
     1321 ccgaccgaaa ggagcgcacg accccatgca tcgattagtc caatttgtta aagacaggat
     1381 atcagtggtc caggctctag ttttgactca acaatatcac cagctgaagc ctatagagta
     1441 cgagccatag ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga
     1501 ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa
     1561 tacataactg agaatagaga agttcagatc aaggtcagga acagatggaa cagggtcgac
     1621 cctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
     1681 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
     1741 tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc
     1801 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg
     1861 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg
     1921 gggctcgtcc gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggta
     1981 agctggctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     2041 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     2101 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
     2161 agtgtagatc cggctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     2221 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     2281 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     2341 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     2401 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     2461 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttac
     2521 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     2581 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
     2641 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2701 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2761 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2821 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2881 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2941 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3001 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3061 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3121 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3181 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3241 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3301 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3361 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3421 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3481 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3541 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3601 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3661 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3721 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3781 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3841 tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc
     3901 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3961 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4021 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4081 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4141 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca
     4201 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4261 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4321 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4381 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4441 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4501 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     4561 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     4621 cgtcttcaag aattagcttg gccattgcat acgttgtatc catatcataa tatgtacatt
     4681 tatattggct catgtccaac attaccgcca tgttgacatt gattattgac tagttattaa
     4741 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     4801 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     4861 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     4921 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     4981 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     5041 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     5101 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     5161 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     5221 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     5281 gtctatataa gcagagctca ataaaagagc ccacaacccc tcactcggcg cgccagtctt
     5341 ccgatagact gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc
     5401 cgaatcgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccacgac
     5461 gggggtcttt catttggggg ctcgtccggg atttggagac ccctgcccag ggaccaccga
     5521 cccaccaccg ggaggtaagc tggccagcaa cttatctgtg tctgtccgat tgtctagtgt
     5581 ctatgtttga tgttatgcgc ctgcgtctgt actagttagc taactagctc tgtatctggc
     5641 ggacccgtgg tggaactgac gagttctgaa cacccggccg caaccctggg agacgtccca
     5701 gggactttgg gggccgtttt tgtggcccga cctgaggaag ggagtcgatg tggaatccga
     5761 ccccgtcagg atatgtggtt ctggtaggag acgagaacct aaaacagttc ccgcctccgt
     5821 ctgaattttt gctttcggtt tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca
     5881 gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagggcc
     5941 agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
     6001 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
     6061 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
     6121 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
     6181 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
     6241 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
     6301 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gccggaattg
     6361 aagatctgtc
//
