LOCUS       sequence-202021-        6367 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 6367)
  AUTHORS   Masutomi K, Possemato R, Wong JM, Currier JL, Tothova Z, Manola JB, 
            Ganesan S, Lansdorp PM, Collins K, Hahn WC
  TITLE     The telomerase reverse transcriptase regulates chromatin state and 
            DNA damage responses.
  JOURNAL   Proc Natl Acad Sci U S A. 2005 Jun 7. 102(23):8222-7.
  PUBMED    15928077
REFERENCE   2  (bases 1 to 6367)
  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: "Proc Natl 
            Acad Sci U S A. 2005 Jun 7. 102(23):8222-7."
COMMENT     Sequence Label: pMKO.1 puro hTERT shRNA
FEATURES             Location/Qualifiers
     source          1..6367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..269
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     21..41
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     192..211
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        357..686
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      537..672
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             696..1295
                     /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(696..715)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1192..1212
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1491..1916
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(2019..2041)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        2182..2511
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2362..2497
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2533..2552
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2633..2650)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2804..3392)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2884..2903)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3563..4354,4355..4423))
                     /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     4186..4205
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4424..4528)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4596..4614
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        4714..5093
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5094..5297
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5248..5268
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5298..5473
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    5536..5735
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5936..6352
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6282..6304
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6320..6336
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 tcgacaaggt cgggcaggaa gagggcctat ttcccatgat tccttcatat ttgcatatac
       61 gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca aagatattag
      121 tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat
      181 gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt
      241 tatatatctt gtggaaagga cgaaacaccg tttcatcagc aagtttggat tcaagagatc
      301 caaacttgct gatgaaattt ttgaattcgc cagcacagtg gtcgaccctg tggaatgtgt
      361 gtcagttagg gtgtggaaag tccccaggct tcccagcagg cagaagtatg caaagcatgc
      421 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
      481 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
      541 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
      601 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
      661 tttttggagg cctaggcttt tgcaaaaagc ttaccatgac cgagtacaag cccacggtgc
      721 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
      781 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
      841 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
      901 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
      961 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     1021 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     1081 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     1141 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     1201 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     1261 gcacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc
     1321 cgaccgaaag gagcgcacga ccccatgcat cgattagtcc aatttgttaa agacaggata
     1381 tcagtggtcc aggctctagt tttgactcaa caatatcacc agctgaagcc tatagagtac
     1441 gagccataga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
     1501 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaaat
     1561 acataactga gaatagagaa gttcagatca aggtcaggaa cagatggaac agggtcgacc
     1621 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
     1681 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
     1741 caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc
     1801 cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt
     1861 tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatttggg
     1921 ggctcgtccg ggatcgggag acccctgccc agggaccacc gacccaccac cgggaggtaa
     1981 gctggctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     2041 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     2101 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     2161 gtgtagatcc ggctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     2221 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     2281 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     2341 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     2401 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     2461 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttact
     2521 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     2581 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
     2641 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     2701 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2761 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2821 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2881 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2941 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3001 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3061 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3121 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3181 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3241 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3301 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3361 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3421 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3481 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3541 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3601 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3661 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3721 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3781 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3841 cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct
     3901 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3961 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4021 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4081 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4141 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac
     4201 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4261 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4321 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     4381 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     4441 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4501 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     4561 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     4621 gtcttcaaga attagcttgg ccattgcata cgttgtatcc atatcataat atgtacattt
     4681 atattggctc atgtccaaca ttaccgccat gttgacattg attattgact agttattaat
     4741 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     4801 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     4861 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     4921 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     4981 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     5041 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     5101 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     5161 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     5221 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     5281 tctatataag cagagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcttc
     5341 cgatagactg cgtcgcccgg gtacccgtat tcccaataaa gcctcttgct gtttgcatcc
     5401 gaatcgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact acccacgacg
     5461 ggggtctttc atttgggggc tcgtccggga tttggagacc cctgcccagg gaccaccgac
     5521 ccaccaccgg gaggtaagct ggccagcaac ttatctgtgt ctgtccgatt gtctagtgtc
     5581 tatgtttgat gttatgcgcc tgcgtctgta ctagttagct aactagctct gtatctggcg
     5641 gacccgtggt ggaactgacg agttctgaac acccggccgc aaccctggga gacgtcccag
     5701 ggactttggg ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac
     5761 cccgtcagga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
     5821 tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag
     5881 catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagggcca
     5941 gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc
     6001 tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg
     6061 gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca
     6121 ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat
     6181 cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc
     6241 taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc
     6301 gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg ccggaattga
     6361 agatctg
//
