LOCUS       sequence-224573-        8545 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 8545)
  AUTHORS   Maida Y, Yasukawa M, Furuuchi M, Lassmann T, Possemato R, Okamoto N,
            Kasim V, Hayashizaki Y, Hahn WC, Masutomi K
  TITLE     An RNA-dependent RNA polymerase formed by TERT and the RMRP RNA.
  JOURNAL   Nature. 2009 Sep 10. 461(7261):230-5.
  PUBMED    19701182
REFERENCE   2  (bases 1 to 8545)
  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: "Nature. 
            2009 Sep 10. 461(7261):230-5."
COMMENT     Sequence Label: pWZL-Blast-Flag-HA-hTERT
FEATURES             Location/Qualifiers
     source          1..8545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..80)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(314..333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(982..1773,1774..1842))
                     /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     1605..1624
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1843..1947)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             1978..2447
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    2512..2711
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             2912..3328
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3258..3280
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3296..3312
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             3357..3380
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3393..3419
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             3393..3419
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3483..6881
                     /codon_start=1
                     /gene="TERT"
                     /product="human telomerase reverse transcriptase, the 
                     catalytic subunit of telomerase"
                     /label=hTERT
                     /note="prevents telomere shortening and allows 
                     postsenescent cells to proliferate beyond crisis (Counter 
                     et al., 1998); also known as EST2"
                     /translation="MPRAPRCRAVRSLLRSHYREVLPLATFVRRLGPQGWRLVQRGDPA
                     AFRALVAQCLVCVPWDARPPPAAPSFRQVSCLKELVARVLQRLCERGAKNVLAFGFALL
                     DGARGGPPEAFTTSVRSYLPNTVTDALRGSGAWGLLLRRVGDDVLVHLLARCALFVLVA
                     PSCAYQVCGPPLYQLGAATQARPPPHASGPRRRLGCERAWNHSVREAGVPLGLPAPGAR
                     RRGGSASRSLPLPKRPRRGAAPEPERTPVGQGSWAHPGRTRGPSDRGFCVVSPARPAEE
                     ATSLEGALSGTRHSHPSVGRQHHAGPPSTSRPPRPWDTPCPPVYAETKHFLYSSGDKEQ
                     LRPSFLLSSLRPSLTGARRLVETIFLGSRPWMPGTPRRLPRLPQRYWQMRPLFLELLGN
                     HAQCPYGVLLKTHCPLRAAVTPAAGVCAREKPQGSVAAPEEEDTDPRRLVQLLRQHSSP
                     WQVYGFVRACLRRLVPPGLWGSRHNERRFLRNTKKFISLGKHAKLSLQELTWKMSVRGC
                     AWLRRSPGVGCVPAAEHRLREEILAKFLHWLMSVYVVELLRSFFYVTETTFQKNRLFFY
                     RKSVWSKLQSIGIRQHLKRVQLRELSEAEVRQHREARPALLTSRLRFIPKPDGLRPIVN
                     MDYVVGARTFRREKRAERLTSRVKALFSVLNYERARRPGLLGASVLGLDDIHRAWRTFV
                     LRVRAQDPPPELYFVKVDVTGAYDTIPQDRLTEVIASIIKPQNTYCVRRYAVVQKAAHG
                     HVRKAFKSHVSTLTDLQPYMRQFVAHLQETSPLRDAVVIEQSSSLNEASSGLFDVFLRF
                     MCHHAVRIRGKSYVQCQGIPQGSILSTLLCSLCYGDMENKLFAGIRRDGLLLRLVDDFL
                     LVTPHLTHAKTFLRTLVRGVPEYGCVVNLRKTVVNFPVEDEALGGTAFVQMPAHGLFPW
                     CGLLLDTRTLEVQSDYSSYARTSIRASLTFNRGFKAGRNMRRKLFGVLRLKCHSLFLDL
                     QVNSLQTVCTNIYKILLLQAYRFHACVLQLPFHQQVWKNPTFFLRVISDTASLCYSILK
                     AKNAGMSLGAKGAAGPLPSEAVQWLCHQAFLLKLTRHRVTYVPLLGSLRTAQTQLSRKL
                     PGTTLTALEAAANPALPSDFKTILD"
     misc_feature    6932..7393
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6988..7005)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7215..7234
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             8009..8478
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
       61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      901 ttatcaaaaa ggatcttcac ctagatcctt ttgcggccgc aaatcaatct aaagtatata
      961 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1021 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     1081 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     1141 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     1201 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     1261 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     1321 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     1381 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     1441 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     1501 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     1561 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     1621 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     1681 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     1741 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     1801 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     1861 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     1921 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgcagcctg
     1981 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     2041 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     2101 gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga
     2161 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     2221 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     2281 agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac
     2341 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     2401 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt
     2461 ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg caagctggcc
     2521 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta tgcgcctgcg
     2581 tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
     2641 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg
     2701 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt
     2761 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa
     2821 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct
     2881 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt
     2941 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa
     3001 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc
     3061 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc
     3121 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt
     3181 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc
     3241 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta
     3301 tccagccctc actccttctc taggcgccgg ccggatcttc cgctcgatga accatggact
     3361 acaaggacga cgatgacaag ctcgatggag gataccccta cgacgtgccc gactacgccg
     3421 gaggactcga ggaattcgtc ctgctgcgca cgtgggaagc cctggccccg gccacccccg
     3481 cgatgccgcg cgctccccgc tgccgagccg tgcgctccct gctgcgcagc cactaccgcg
     3541 aggtgctgcc gctggccacg ttcgtgcggc gcctggggcc ccagggctgg cggctggtgc
     3601 agcgcgggga cccggcggct ttccgcgcgc tggtggccca gtgcctggtg tgcgtgccct
     3661 gggacgcacg gccgcccccc gccgccccct ccttccgcca ggtgtcctgc ctgaaggagc
     3721 tggtggcccg agtgctgcag aggctgtgcg agcgcggcgc gaagaacgtg ctggccttcg
     3781 gcttcgcgct gctggacggg gcccgcgggg gcccccccga ggccttcacc accagcgtgc
     3841 gcagctacct gcccaacacg gtgaccgacg cactgcgggg gagcggggcg tgggggctgc
     3901 tgctgcgccg cgtgggcgac gacgtgctgg ttcacctgct ggcacgctgc gcgctctttg
     3961 tgctggtggc tcccagctgc gcctaccagg tgtgcgggcc gccgctgtac cagctcggcg
     4021 ctgccactca ggcccggccc ccgccacacg ctagtggacc ccgaaggcgt ctgggatgcg
     4081 aacgggcctg gaaccatagc gtcagggagg ccggggtccc cctgggcctg ccagccccgg
     4141 gtgcgaggag gcgcgggggc agtgccagcc gaagtctgcc gttgcccaag aggcccaggc
     4201 gtggcgctgc ccctgagccg gagcggacgc ccgttgggca ggggtcctgg gcccacccgg
     4261 gcaggacgcg tggaccgagt gaccgtggtt tctgtgtggt gtcacctgcc agacccgccg
     4321 aagaagccac ctctttggag ggtgcgctct ctggcacgcg ccactcccac ccatccgtgg
     4381 gccgccagca ccacgcgggc cccccatcca catcgcggcc accacgtccc tgggacacgc
     4441 cttgtccccc ggtgtacgcc gagaccaagc acttcctcta ctcctcaggc gacaaggagc
     4501 agctgcggcc ctccttccta ctcagctctc tgaggcccag cctgactggc gctcggaggc
     4561 tcgtggagac catctttctg ggttccaggc cctggatgcc agggactccc cgcaggttgc
     4621 cccgcctgcc ccagcgctac tggcaaatgc ggcccctgtt tctggagctg cttgggaacc
     4681 acgcgcagtg cccctacggg gtgctcctca agacgcactg cccgctgcga gctgcggtca
     4741 ccccagcagc cggtgtctgt gcccgggaga agccccaggg ctctgtggcg gcccccgagg
     4801 aggaggacac agacccccgt cgcctggtgc agctgctccg ccagcacagc agcccctggc
     4861 aggtgtacgg cttcgtgcgg gcctgcctgc gccggctggt gcccccaggc ctctggggct
     4921 ccaggcacaa cgaacgccgc ttcctcagga acaccaagaa gttcatctcc ctggggaagc
     4981 atgccaagct ctcgctgcag gagctgacgt ggaagatgag cgtgcggggc tgcgcttggc
     5041 tgcgcaggag cccaggggtt ggctgtgttc cggccgcaga gcaccgtctg cgtgaggaga
     5101 tcctggccaa gttcctgcac tggctgatga gtgtgtacgt cgtcgagctg ctcaggtctt
     5161 tcttttatgt cacggagacc acgtttcaaa agaacaggct ctttttctac cggaagagtg
     5221 tctggagcaa gttgcaaagc attggaatca gacagcactt gaagagggtg cagctgcggg
     5281 agctgtcgga agcagaggtc aggcagcatc gggaagccag gcccgccctg ctgacgtcca
     5341 gactccgctt catccccaag cctgacgggc tgcggccgat tgtgaacatg gactacgtcg
     5401 tgggagccag aacgttccgc agagaaaaga gggccgagcg tctcacctcg agggtgaagg
     5461 cactgttcag cgtgctcaac tacgagcggg cgcggcgccc cggcctcctg ggcgcctctg
     5521 tgctgggcct ggacgatatc cacagggcct ggcgcacctt cgtgctgcgt gtgcgggccc
     5581 aggacccgcc gcctgagctg tactttgtca aggtggatgt gacgggcgcg tacgacacca
     5641 tcccccagga caggctcacg gaggtcatcg ccagcatcat caaaccccag aacacgtact
     5701 gcgtgcgtcg gtatgccgtg gtccagaagg ccgcccatgg gcacgtccgc aaggccttca
     5761 agagccacgt ctctaccttg acagacctcc agccgtacat gcgacagttc gtggctcacc
     5821 tgcaggagac cagcccgctg agggatgccg tcgtcatcga gcagagctcc tccctgaatg
     5881 aggccagcag tggcctcttc gacgtcttcc tacgcttcat gtgccaccac gccgtgcgca
     5941 tcaggggcaa gtcctacgtc cagtgccagg ggatcccgca gggctccatc ctctccacgc
     6001 tgctctgcag cctgtgctac ggcgacatgg agaacaagct gtttgcgggg attcggcggg
     6061 acgggctgct cctgcgtttg gtggatgatt tcttgttggt gacacctcac ctcacccacg
     6121 cgaaaacctt cctcaggacc ctggtccgag gtgtccctga gtatggctgc gtggtgaact
     6181 tgcggaagac agtggtgaac ttccctgtag aagacgaggc cctgggtggc acggcttttg
     6241 ttcagatgcc ggcccacggc ctattcccct ggtgcggcct gctgctggat acccggaccc
     6301 tggaggtgca gagcgactac tccagctatg cccggacctc catcagagcc agtctcacct
     6361 tcaaccgcgg cttcaaggct gggaggaaca tgcgtcgcaa actctttggg gtcttgcggc
     6421 tgaagtgtca cagcctgttt ctggatttgc aggtgaacag cctccagacg gtgtgcacca
     6481 acatctacaa gatcctcctg ctgcaggcgt acaggtttca cgcatgtgtg ctgcagctcc
     6541 catttcatca gcaagtttgg aagaacccca catttttcct gcgcgtcatc tctgacacgg
     6601 cctccctctg ctactccatc ctgaaagcca agaacgcagg gatgtcgctg ggggccaagg
     6661 gcgccgccgg ccctctgccc tccgaggccg tgcagtggct gtgccaccaa gcattcctgc
     6721 tcaagctgac tcgacaccgt gtcacctacg tgccactcct ggggtcactc aggacagccc
     6781 agacgcagct gagtcggaag ctcccgggga cgacgctgac tgccctggag gccgcagcca
     6841 acccggcact gccctcagac ttcaagacca tcctggactg agtcgaccaa ttccggttat
     6901 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     6961 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
     7021 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc
     7081 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     7141 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     7201 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     7261 aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt
     7321 agtcgaggtt aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     7381 aacacgatga taataccatg aaaacattta acatttctca acaggatcta gaattagtag
     7441 aagtagcgac agagaagatt acaatgcttt atgaggataa taaacatcat gtgggagcgg
     7501 caattcgtac gaaaacagga gaaatcattt cggcagtaca tattgaagcg tatataggac
     7561 gagtaactgt ttgtgcagaa gccattgcga ttggtagtgc agtttcgaat ggacaaaagg
     7621 attttgacac gattgtagct gttagacacc cttattctga cgaagtagat agaagtattc
     7681 gagtggtaag tccttgtggt atgtgtaggg agttgatttc agactatgca ccagattgtt
     7741 ttgtgttaat agaaatgaat ggcaagttag tcaaaactac gattgaagaa ctcattccac
     7801 tcaaatatac ccgaaattaa aagttttacc ataccaagct catcgataaa ataaaagatt
     7861 ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta
     7921 gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat agagaagttc
     7981 agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat atctgtggta
     8041 agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata tgggccaaac
     8101 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag
     8161 atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag
     8221 gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
     8281 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggggcgcc
     8341 agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt
     8401 gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc
     8461 gtcagcgggg gtctttcaca tgcagcatgt atcaaaatta atttggtttt ttttcttaag
     8521 tatttacatt aaatggccat agttg
//
