LOCUS       sequence-249044-        8518 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 8518)
  AUTHORS   Hahn WC, Stewart SA, Brooks MW, York SG, Eaton E, Kurachi A, 
            Beijersbergen RL, Knoll JH, Meyerson M, Weinberg RA
  TITLE     Inhibition of telomerase limits the growth of human cancer cells.
  JOURNAL   Nat Med 1999 Oct;5(10):1164-70.
  PUBMED    10502820
REFERENCE   2  (bases 1 to 8518)
  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: "Nat Med"; 
            date: "1999-10"; volume: "5(10)"; pages: "1164-70"
COMMENT     Sequence Label: pBABE-puro-DN-hTERT
FEATURES             Location/Qualifiers
     source          1..8518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2..21)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(670..1461,1462..1530))
                     /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     1293..1312
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1531..1635)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             1666..2135
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    2200..2399
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             2600..3016
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2946..2968
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2984..3000
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             3096..6494
                     /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
                     LRVRAQDPPPELYFVKVAITGAYDTIPQDRLTEVIASIIKPQNTYCVRRYAVVQKAAHG
                     HVRKAFKSHVSTLTDLQPYMRQFVAHLQETSPLRDAVVIEQSSSLNEASSGLFDVFLRF
                     MCHHAVRIRGKSYVQCQGIPQGSILSTLLCSLCYGDMENKLFAGIRRDGLLLRLVDDFL
                     LVTPHLTHAKTFLRTLVRGVPEYGCVVNLRKTVVNFPVEDEALGGTAFVQMPAHGLFPW
                     CGLLLDTRTLEVQSDYSSYARTSIRASLTFNRGFKAGRNMRRKLFGVLRLKCHSLFLDL
                     QVNSLQTVCTNIYKILLLQAYRFHACVLQLPFHQQVWKNPTFFLRVISDTASLCYSILK
                     AKNAGMSLGAKGAAGPLPSEAVQWLCHQAFLLKLTRHRVTYVPLLGSLRTAQTQLSRKL
                     PGTTLTALEAAANPALPSDFKTILD"
     primer_bind     complement(6506..6526)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6511..6840
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6691..6826
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6850..7449
                     /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(6850..6869)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7346..7366
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             7670..8139
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(8269..8286)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(8440..8518,1..510))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
       61 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      121 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      181 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      241 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      301 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      361 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      421 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      481 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      541 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      601 atcttcacct agatcctttt gcggccgcaa atcaatctaa agtatatatg agtaaacttg
      661 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
      721 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
      781 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
      841 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
      901 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
      961 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     1021 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     1081 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     1141 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     1201 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     1261 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     1321 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     1381 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     1441 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     1501 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     1561 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     1621 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gcagcctgaa tatgggccaa
     1681 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag
     1741 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     1801 gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt
     1861 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     1921 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     1981 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
     2041 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     2101 tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga
     2161 gacccctgcc cagggaccac cgacccacca ccgggaggca agctggccag caacttatct
     2221 gtgtctgtcc gattgtctag tgtctatgac tgattttatg cgcctgcgtc ggtactagtt
     2281 agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttct gaacacccgg
     2341 ccgcaaccct gggagacgtc ccagggactt tgggggccgt ttttgtggcc cgacctgagg
     2401 aagggagtcg atgtggaatc cgaccccgtc aggatatgtg gttctggtag gagacgagaa
     2461 cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc gaagccgcgc
     2521 gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt ctctgtctga ctgtgtttct
     2581 gtatttgtct gaaaattagg gccagactgt taccactccc ttaagtttga ccttaggtca
     2641 ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga agagacgttg
     2701 ggttaccttc tgctctgcag aatggccaac ctttaacgtc ggatggccgc gagacggcac
     2761 ctttaaccga gacctcatca cccaggttaa gatcaaggtc ttttcacctg gcccgcatgg
     2821 acacccagac caggtcccct acatcgtgac ctgggaagcc ttggcttttg acccccctcc
     2881 ctgggtcaag ccctttgtac accctaagcc tccgcctcct cttcctccat ccgccccgtc
     2941 tctccccctt gaacctcctc gttcgacccc gcctcgatcc tccctttatc cagccctcac
     3001 tccttctcta ggcgccggcc ggatcccagt gtggtggtac gtaggaattc gtcctgctgc
     3061 gcacgtggga agccctggcc ccggccaccc ccgcgatgcc gcgcgctccc cgctgccgag
     3121 ccgtgcgctc cctgctgcgc agccactacc gcgaggtgct gccgctggcc acgttcgtgc
     3181 ggcgcctggg gccccagggc tggcggctgg tgcagcgcgg ggacccggcg gctttccgcg
     3241 cgctggtggc ccagtgcctg gtgtgcgtgc cctgggacgc acggccgccc cccgccgccc
     3301 cctccttccg ccaggtgtcc tgcctgaagg agctggtggc ccgagtgctg cagaggctgt
     3361 gcgagcgcgg cgcgaagaac gtgctggcct tcggcttcgc gctgctggac ggggcccgcg
     3421 ggggcccccc cgaggccttc accaccagcg tgcgcagcta cctgcccaac acggtgaccg
     3481 acgcactgcg ggggagcggg gcgtgggggc tgctgctgcg ccgcgtgggc gacgacgtgc
     3541 tggttcacct gctggcacgc tgcgcgctct ttgtgctggt ggctcccagc tgcgcctacc
     3601 aggtgtgcgg gccgccgctg taccagctcg gcgctgccac tcaggcccgg cccccgccac
     3661 acgctagtgg accccgaagg cgtctgggat gcgaacgggc ctggaaccat agcgtcaggg
     3721 aggccggggt ccccctgggc ctgccagccc cgggtgcgag gaggcgcggg ggcagtgcca
     3781 gccgaagtct gccgttgccc aagaggccca ggcgtggcgc tgcccctgag ccggagcgga
     3841 cgcccgttgg gcaggggtcc tgggcccacc cgggcaggac gcgtggaccg agtgaccgtg
     3901 gtttctgtgt ggtgtcacct gccagacccg ccgaagaagc cacctctttg gagggtgcgc
     3961 tctctggcac gcgccactcc cacccatccg tgggccgcca gcaccacgcg ggccccccat
     4021 ccacatcgcg gccaccacgt ccctgggaca cgccttgtcc cccggtgtac gccgagacca
     4081 agcacttcct ctactcctca ggcgacaagg agcagctgcg gccctccttc ctactcagct
     4141 ctctgaggcc cagcctgact ggcgctcgga ggctcgtgga gaccatcttt ctgggttcca
     4201 ggccctggat gccagggact ccccgcaggt tgccccgcct gccccagcgc tactggcaaa
     4261 tgcggcccct gtttctggag ctgcttggga accacgcgca gtgcccctac ggggtgctcc
     4321 tcaagacgca ctgcccgctg cgagctgcgg tcaccccagc agccggtgtc tgtgcccggg
     4381 agaagcccca gggctctgtg gcggcccccg aggaggagga cacagacccc cgtcgcctgg
     4441 tgcagctgct ccgccagcac agcagcccct ggcaggtgta cggcttcgtg cgggcctgcc
     4501 tgcgccggct ggtgccccca ggcctctggg gctccaggca caacgaacgc cgcttcctca
     4561 ggaacaccaa gaagttcatc tccctgggga agcatgccaa gctctcgctg caggagctga
     4621 cgtggaagat gagcgtgcgg ggctgcgctt ggctgcgcag gagcccaggg gttggctgtg
     4681 ttccggccgc agagcaccgt ctgcgtgagg agatcctggc caagttcctg cactggctga
     4741 tgagtgtgta cgtcgtcgag ctgctcaggt ctttctttta tgtcacggag accacgtttc
     4801 aaaagaacag gctctttttc taccggaaga gtgtctggag caagttgcaa agcattggaa
     4861 tcagacagca cttgaagagg gtgcagctgc gggagctgtc ggaagcagag gtcaggcagc
     4921 atcgggaagc caggcccgcc ctgctgacgt ccagactccg cttcatcccc aagcctgacg
     4981 ggctgcggcc gattgtgaac atggactacg tcgtgggagc cagaacgttc cgcagagaaa
     5041 agagggccga gcgtctcacc tcgagggtga aggcactgtt cagcgtgctc aactacgagc
     5101 gggcgcggcg ccccggcctc ctgggcgcct ctgtgctggg cctggacgat atccacaggg
     5161 cctggcgcac cttcgtgctg cgtgtgcggg cccaggaccc gccgcctgag ctgtactttg
     5221 tcaaggtcgc gatcacgggc gcgtacgaca ccatccccca ggacaggctc acggaggtca
     5281 tcgccagcat catcaaaccc cagaacacgt actgcgtgcg tcggtatgcc gtggtccaga
     5341 aggccgccca tgggcacgtc cgcaaggcct tcaagagcca cgtctctacc ttgacagacc
     5401 tccagccgta catgcgacag ttcgtggctc acctgcagga gaccagcccg ctgagggatg
     5461 ccgtcgtcat cgagcagagc tcctccctga atgaggccag cagtggcctc ttcgacgtct
     5521 tcctacgctt catgtgccac cacgccgtgc gcatcagggg caagtcctac gtccagtgcc
     5581 aggggatccc gcagggctcc atcctctcca cgctgctctg cagcctgtgc tacggcgaca
     5641 tggagaacaa gctgtttgcg gggattcggc gggacgggct gctcctgcgt ttggtggatg
     5701 atttcttgtt ggtgacacct cacctcaccc acgcgaaaac cttcctcagg accctggtcc
     5761 gaggtgtccc tgagtatggc tgcgtggtga acttgcggaa gacagtggtg aacttccctg
     5821 tagaagacga ggccctgggt ggcacggctt ttgttcagat gccggcccac ggcctattcc
     5881 cctggtgcgg cctgctgctg gatacccgga ccctggaggt gcagagcgac tactccagct
     5941 atgcccggac ctccatcaga gccagtctca ccttcaaccg cggcttcaag gctgggagga
     6001 acatgcgtcg caaactcttt ggggtcttgc ggctgaagtg tcacagcctg tttctggatt
     6061 tgcaggtgaa cagcctccag acggtgtgca ccaacatcta caagatcctc ctgctgcagg
     6121 cgtacaggtt tcacgcatgt gtgctgcagc tcccatttca tcagcaagtt tggaagaacc
     6181 ccacattttt cctgcgcgtc atctctgaca cggcctccct ctgctactcc atcctgaaag
     6241 ccaagaacgc agggatgtcg ctgggggcca agggcgccgc cggccctctg ccctccgagg
     6301 ccgtgcagtg gctgtgccac caagcattcc tgctcaagct gactcgacac cgtgtcacct
     6361 acgtgccact cctggggtca ctcaggacag cccagacgca gctgagtcgg aagctcccgg
     6421 ggacgacgct gactgccctg gaggccgcag ccaacccggc actgccctca gacttcaaga
     6481 ccatcctgga ctgagtcgac cctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     6541 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     6601 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     6661 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     6721 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     6781 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     6841 aagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     6901 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     6961 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     7021 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     7081 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     7141 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     7201 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     7261 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     7321 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     7381 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     7441 ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc acgaccccat
     7501 gcatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc
     7561 acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaaataca
     7621 taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc tgaatatggg
     7681 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatgga
     7741 acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     7801 ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga gaaccatcag
     7861 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     7921 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     7981 caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat
     8041 ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct
     8101 cctctgagtg attgactacc cgtcagcggg ggtctttcac atgcagcatg tatcaaaatt
     8161 aatttggttt tttttcttaa gtatttacat taaatggcca tagttgcatt aatgaatcgg
     8221 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     8281 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     8341 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     8401 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     8461 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggacta
//
