LOCUS       sequence-373046-        8518 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of hTERT. Used to create immortalized cell 
            lines..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8518)
  AUTHORS   Counter CM, Hahn WC, Wei W, Caddle SD, Beijersbergen RL, Lansdorp 
            PM, Sedivy JM, Weinberg RA
  TITLE     Dissociation among in vitro telomerase activity, telomere 
            maintenance, and cellular immortalization.
  JOURNAL   Proc Natl Acad Sci U S A 1998 Dec 8;95(25):14723-8.
  PUBMED    9843956
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: "Proc Natl 
            Acad Sci U S A"; date: "1998-12-8"; volume: "95(25)"; pages: 
            "14723-8"
COMMENT     Sequence Label: pBABE-puro-hTERT
FEATURES             Location/Qualifiers
     source          1..8518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             101..570
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    635..834
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1035..1451
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1381..1403
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1419..1435
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1531..4929
                     /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"
     primer_bind     complement(4941..4961)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4946..5275
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5126..5261
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5285..5884
                     /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(5285..5304)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5781..5801
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6105..6574
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6704..6721)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6875..7463)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6955..6974)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7623..8414,8415..8483))
                     /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     8246..8265
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8484..8518,1..70))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
       61 gggttccgcg cacatttccc cgaaaagtgc cacctgcagc ctgaatatgg gccaaacagg
      121 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg aacagctgaa
      181 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
      241 gatggtcccc agatgcggtc cagccctcag cagtttctag agaaccatca gatgtttcca
      301 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
      361 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc
      421 actcggggcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac
      481 cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt
      541 gattgactac ccgtcagcgg gggtctttca tttgggggct cgtccgggat cgggagaccc
      601 ctgcccaggg accaccgacc caccaccggg aggcaagctg gccagcaact tatctgtgtc
      661 tgtccgattg tctagtgtct atgactgatt ttatgcgcct gcgtcggtac tagttagcta
      721 actagctctg tatctggcgg acccgtggtg gaactgacga gttctgaaca cccggccgca
      781 accctgggag acgtcccagg gactttgggg gccgtttttg tggcccgacc tgaggaaggg
      841 agtcgatgtg gaatccgacc ccgtcaggat atgtggttct ggtaggagac gagaacctaa
      901 aacagttccc gcctccgtct gaatttttgc tttcggtttg gaaccgaagc cgcgcgtctt
      961 gtctgctgca gcgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt
     1021 tgtctgaaaa ttagggccag actgttacca ctcccttaag tttgacctta ggtcactgga
     1081 aagatgtcga gcggatcgct cacaaccagt cggtagatgt caagaagaga cgttgggtta
     1141 ccttctgctc tgcagaatgg ccaaccttta acgtcggatg gccgcgagac ggcaccttta
     1201 accgagacct catcacccag gttaagatca aggtcttttc acctggcccg catggacacc
     1261 cagaccaggt cccctacatc gtgacctggg aagccttggc ttttgacccc cctccctggg
     1321 tcaagccctt tgtacaccct aagcctccgc ctcctcttcc tccatccgcc ccgtctctcc
     1381 cccttgaacc tcctcgttcg accccgcctc gatcctccct ttatccagcc ctcactcctt
     1441 ctctaggcgc cggccggatc ccagtgtggt ggtacgtagg aattcgtcct gctgcgcacg
     1501 tgggaagccc tggccccggc cacccccgcg atgccgcgcg ctccccgctg ccgagccgtg
     1561 cgctccctgc tgcgcagcca ctaccgcgag gtgctgccgc tggccacgtt cgtgcggcgc
     1621 ctggggcccc agggctggcg gctggtgcag cgcggggacc cggcggcttt ccgcgcgctg
     1681 gtggcccagt gcctggtgtg cgtgccctgg gacgcacggc cgccccccgc cgccccctcc
     1741 ttccgccagg tgtcctgcct gaaggagctg gtggcccgag tgctgcagag gctgtgcgag
     1801 cgcggcgcga agaacgtgct ggccttcggc ttcgcgctgc tggacggggc ccgcgggggc
     1861 ccccccgagg ccttcaccac cagcgtgcgc agctacctgc ccaacacggt gaccgacgca
     1921 ctgcggggga gcggggcgtg ggggctgctg ctgcgccgcg tgggcgacga cgtgctggtt
     1981 cacctgctgg cacgctgcgc gctctttgtg ctggtggctc ccagctgcgc ctaccaggtg
     2041 tgcgggccgc cgctgtacca gctcggcgct gccactcagg cccggccccc gccacacgct
     2101 agtggacccc gaaggcgtct gggatgcgaa cgggcctgga accatagcgt cagggaggcc
     2161 ggggtccccc tgggcctgcc agccccgggt gcgaggaggc gcgggggcag tgccagccga
     2221 agtctgccgt tgcccaagag gcccaggcgt ggcgctgccc ctgagccgga gcggacgccc
     2281 gttgggcagg ggtcctgggc ccacccgggc aggacgcgtg gaccgagtga ccgtggtttc
     2341 tgtgtggtgt cacctgccag acccgccgaa gaagccacct ctttggaggg tgcgctctct
     2401 ggcacgcgcc actcccaccc atccgtgggc cgccagcacc acgcgggccc cccatccaca
     2461 tcgcggccac cacgtccctg ggacacgcct tgtcccccgg tgtacgccga gaccaagcac
     2521 ttcctctact cctcaggcga caaggagcag ctgcggccct ccttcctact cagctctctg
     2581 aggcccagcc tgactggcgc tcggaggctc gtggagacca tctttctggg ttccaggccc
     2641 tggatgccag ggactccccg caggttgccc cgcctgcccc agcgctactg gcaaatgcgg
     2701 cccctgtttc tggagctgct tgggaaccac gcgcagtgcc cctacggggt gctcctcaag
     2761 acgcactgcc cgctgcgagc tgcggtcacc ccagcagccg gtgtctgtgc ccgggagaag
     2821 ccccagggct ctgtggcggc ccccgaggag gaggacacag acccccgtcg cctggtgcag
     2881 ctgctccgcc agcacagcag cccctggcag gtgtacggct tcgtgcgggc ctgcctgcgc
     2941 cggctggtgc ccccaggcct ctggggctcc aggcacaacg aacgccgctt cctcaggaac
     3001 accaagaagt tcatctccct ggggaagcat gccaagctct cgctgcagga gctgacgtgg
     3061 aagatgagcg tgcggggctg cgcttggctg cgcaggagcc caggggttgg ctgtgttccg
     3121 gccgcagagc accgtctgcg tgaggagatc ctggccaagt tcctgcactg gctgatgagt
     3181 gtgtacgtcg tcgagctgct caggtctttc ttttatgtca cggagaccac gtttcaaaag
     3241 aacaggctct ttttctaccg gaagagtgtc tggagcaagt tgcaaagcat tggaatcaga
     3301 cagcacttga agagggtgca gctgcgggag ctgtcggaag cagaggtcag gcagcatcgg
     3361 gaagccaggc ccgccctgct gacgtccaga ctccgcttca tccccaagcc tgacgggctg
     3421 cggccgattg tgaacatgga ctacgtcgtg ggagccagaa cgttccgcag agaaaagagg
     3481 gccgagcgtc tcacctcgag ggtgaaggca ctgttcagcg tgctcaacta cgagcgggcg
     3541 cggcgccccg gcctcctggg cgcctctgtg ctgggcctgg acgatatcca cagggcctgg
     3601 cgcaccttcg tgctgcgtgt gcgggcccag gacccgccgc ctgagctgta ctttgtcaag
     3661 gtggatgtga cgggcgcgta cgacaccatc ccccaggaca ggctcacgga ggtcatcgcc
     3721 agcatcatca aaccccagaa cacgtactgc gtgcgtcggt atgccgtggt ccagaaggcc
     3781 gcccatgggc acgtccgcaa ggccttcaag agccacgtct ctaccttgac agacctccag
     3841 ccgtacatgc gacagttcgt ggctcacctg caggagacca gcccgctgag ggatgccgtc
     3901 gtcatcgagc agagctcctc cctgaatgag gccagcagtg gcctcttcga cgtcttccta
     3961 cgcttcatgt gccaccacgc cgtgcgcatc aggggcaagt cctacgtcca gtgccagggg
     4021 atcccgcagg gctccatcct ctccacgctg ctctgcagcc tgtgctacgg cgacatggag
     4081 aacaagctgt ttgcggggat tcggcgggac gggctgctcc tgcgtttggt ggatgatttc
     4141 ttgttggtga cacctcacct cacccacgcg aaaaccttcc tcaggaccct ggtccgaggt
     4201 gtccctgagt atggctgcgt ggtgaacttg cggaagacag tggtgaactt ccctgtagaa
     4261 gacgaggccc tgggtggcac ggcttttgtt cagatgccgg cccacggcct attcccctgg
     4321 tgcggcctgc tgctggatac ccggaccctg gaggtgcaga gcgactactc cagctatgcc
     4381 cggacctcca tcagagccag tctcaccttc aaccgcggct tcaaggctgg gaggaacatg
     4441 cgtcgcaaac tctttggggt cttgcggctg aagtgtcaca gcctgtttct ggatttgcag
     4501 gtgaacagcc tccagacggt gtgcaccaac atctacaaga tcctcctgct gcaggcgtac
     4561 aggtttcacg catgtgtgct gcagctccca tttcatcagc aagtttggaa gaaccccaca
     4621 tttttcctgc gcgtcatctc tgacacggcc tccctctgct actccatcct gaaagccaag
     4681 aacgcaggga tgtcgctggg ggccaagggc gccgccggcc ctctgccctc cgaggccgtg
     4741 cagtggctgt gccaccaagc attcctgctc aagctgactc gacaccgtgt cacctacgtg
     4801 ccactcctgg ggtcactcag gacagcccag acgcagctga gtcggaagct cccggggacg
     4861 acgctgactg ccctggaggc cgcagccaac ccggcactgc cctcagactt caagaccatc
     4921 ctggactgag tcgaccctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4981 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     5041 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5101 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     5161 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     5221 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     5281 taccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
     5341 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
     5401 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     5461 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     5521 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     5581 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     5641 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     5701 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     5761 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     5821 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     5881 ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc
     5941 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg
     6001 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact
     6061 gagaatagag aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa
     6121 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc
     6181 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     6241 aacagatggt ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt
     6301 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
     6361 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
     6421 cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat
     6481 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct
     6541 gagtgattga ctacccgtca gcgggggtct ttcacatgca gcatgtatca aaattaattt
     6601 ggtttttttt cttaagtatt tacattaaat ggccatagtt gcattaatga atcggccaac
     6661 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     6721 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6781 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6841 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6901 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6961 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7021 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     7081 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7141 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7201 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7261 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     7321 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7381 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7441 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7501 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7561 cctagatcct tttgcggccg caaatcaatc taaagtatat atgagtaaac ttggtctgac
     7621 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7681 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7741 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7801 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7861 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7921 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7981 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     8041 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     8101 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8161 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8221 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     8281 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8341 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8401 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8461 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatc
//
