LOCUS       Exported                8518 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Retroviral expression of hTERT. Used to create immortalized cell 
            lines..
ACCESSION   .
VERSION     .
KEYWORDS    pBABE-puro-hTERT
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 May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8518
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             24..493
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    558..757
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             958..1374
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1304..1326
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1342..1358
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1454..4852
                     /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(4864..4884)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4869..5198
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5049..5184
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5111..5130
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5208..5807
                     /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(5208..5227)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5704..5724
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6028..6497
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6627..6644)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6798..7386)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6878..6897)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7546..8406)
                     /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     8169..8188
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8407..8511)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccccgaaaag tgccacctgc agcctgaata tgggccaaac aggatatctg tggtaagcag
       61 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
      121 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
      181 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
      241 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
      301 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc
      361 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
      421 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
      481 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
      541 acccaccacc gggaggcaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
      601 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg
      661 cggacccgtg gtggaactga cgagttctga acacccggcc gcaaccctgg gagacgtccc
      721 agggactttg ggggccgttt ttgtggcccg acctgaggaa gggagtcgat gtggaatccg
      781 accccgtcag gatatgtggt tctggtagga gacgagaacc taaaacagtt cccgcctccg
      841 tctgaatttt tgctttcggt ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc
      901 agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattagggc
      961 cagactgtta ccactccctt aagtttgacc ttaggtcact ggaaagatgt cgagcggatc
     1021 gctcacaacc agtcggtaga tgtcaagaag agacgttggg ttaccttctg ctctgcagaa
     1081 tggccaacct ttaacgtcgg atggccgcga gacggcacct ttaaccgaga cctcatcacc
     1141 caggttaaga tcaaggtctt ttcacctggc ccgcatggac acccagacca ggtcccctac
     1201 atcgtgacct gggaagcctt ggcttttgac ccccctccct gggtcaagcc ctttgtacac
     1261 cctaagcctc cgcctcctct tcctccatcc gccccgtctc tcccccttga acctcctcgt
     1321 tcgaccccgc ctcgatcctc cctttatcca gccctcactc cttctctagg cgccggccgg
     1381 atcccagtgt ggtggtacgt aggaattcgt cctgctgcgc acgtgggaag ccctggcccc
     1441 ggccaccccc gcgatgccgc gcgctccccg ctgccgagcc gtgcgctccc tgctgcgcag
     1501 ccactaccgc gaggtgctgc cgctggccac gttcgtgcgg cgcctggggc cccagggctg
     1561 gcggctggtg cagcgcgggg acccggcggc tttccgcgcg ctggtggccc agtgcctggt
     1621 gtgcgtgccc tgggacgcac ggccgccccc cgccgccccc tccttccgcc aggtgtcctg
     1681 cctgaaggag ctggtggccc gagtgctgca gaggctgtgc gagcgcggcg cgaagaacgt
     1741 gctggccttc ggcttcgcgc tgctggacgg ggcccgcggg ggcccccccg aggccttcac
     1801 caccagcgtg cgcagctacc tgcccaacac ggtgaccgac gcactgcggg ggagcggggc
     1861 gtgggggctg ctgctgcgcc gcgtgggcga cgacgtgctg gttcacctgc tggcacgctg
     1921 cgcgctcttt gtgctggtgg ctcccagctg cgcctaccag gtgtgcgggc cgccgctgta
     1981 ccagctcggc gctgccactc aggcccggcc cccgccacac gctagtggac cccgaaggcg
     2041 tctgggatgc gaacgggcct ggaaccatag cgtcagggag gccggggtcc ccctgggcct
     2101 gccagccccg ggtgcgagga ggcgcggggg cagtgccagc cgaagtctgc cgttgcccaa
     2161 gaggcccagg cgtggcgctg cccctgagcc ggagcggacg cccgttgggc aggggtcctg
     2221 ggcccacccg ggcaggacgc gtggaccgag tgaccgtggt ttctgtgtgg tgtcacctgc
     2281 cagacccgcc gaagaagcca cctctttgga gggtgcgctc tctggcacgc gccactccca
     2341 cccatccgtg ggccgccagc accacgcggg ccccccatcc acatcgcggc caccacgtcc
     2401 ctgggacacg ccttgtcccc cggtgtacgc cgagaccaag cacttcctct actcctcagg
     2461 cgacaaggag cagctgcggc cctccttcct actcagctct ctgaggccca gcctgactgg
     2521 cgctcggagg ctcgtggaga ccatctttct gggttccagg ccctggatgc cagggactcc
     2581 ccgcaggttg ccccgcctgc cccagcgcta ctggcaaatg cggcccctgt ttctggagct
     2641 gcttgggaac cacgcgcagt gcccctacgg ggtgctcctc aagacgcact gcccgctgcg
     2701 agctgcggtc accccagcag ccggtgtctg tgcccgggag aagccccagg gctctgtggc
     2761 ggcccccgag gaggaggaca cagacccccg tcgcctggtg cagctgctcc gccagcacag
     2821 cagcccctgg caggtgtacg gcttcgtgcg ggcctgcctg cgccggctgg tgcccccagg
     2881 cctctggggc tccaggcaca acgaacgccg cttcctcagg aacaccaaga agttcatctc
     2941 cctggggaag catgccaagc tctcgctgca ggagctgacg tggaagatga gcgtgcgggg
     3001 ctgcgcttgg ctgcgcagga gcccaggggt tggctgtgtt ccggccgcag agcaccgtct
     3061 gcgtgaggag atcctggcca agttcctgca ctggctgatg agtgtgtacg tcgtcgagct
     3121 gctcaggtct ttcttttatg tcacggagac cacgtttcaa aagaacaggc tctttttcta
     3181 ccggaagagt gtctggagca agttgcaaag cattggaatc agacagcact tgaagagggt
     3241 gcagctgcgg gagctgtcgg aagcagaggt caggcagcat cgggaagcca ggcccgccct
     3301 gctgacgtcc agactccgct tcatccccaa gcctgacggg ctgcggccga ttgtgaacat
     3361 ggactacgtc gtgggagcca gaacgttccg cagagaaaag agggccgagc gtctcacctc
     3421 gagggtgaag gcactgttca gcgtgctcaa ctacgagcgg gcgcggcgcc ccggcctcct
     3481 gggcgcctct gtgctgggcc tggacgatat ccacagggcc tggcgcacct tcgtgctgcg
     3541 tgtgcgggcc caggacccgc cgcctgagct gtactttgtc aaggtggatg tgacgggcgc
     3601 gtacgacacc atcccccagg acaggctcac ggaggtcatc gccagcatca tcaaacccca
     3661 gaacacgtac tgcgtgcgtc ggtatgccgt ggtccagaag gccgcccatg ggcacgtccg
     3721 caaggccttc aagagccacg tctctacctt gacagacctc cagccgtaca tgcgacagtt
     3781 cgtggctcac ctgcaggaga ccagcccgct gagggatgcc gtcgtcatcg agcagagctc
     3841 ctccctgaat gaggccagca gtggcctctt cgacgtcttc ctacgcttca tgtgccacca
     3901 cgccgtgcgc atcaggggca agtcctacgt ccagtgccag gggatcccgc agggctccat
     3961 cctctccacg ctgctctgca gcctgtgcta cggcgacatg gagaacaagc tgtttgcggg
     4021 gattcggcgg gacgggctgc tcctgcgttt ggtggatgat ttcttgttgg tgacacctca
     4081 cctcacccac gcgaaaacct tcctcaggac cctggtccga ggtgtccctg agtatggctg
     4141 cgtggtgaac ttgcggaaga cagtggtgaa cttccctgta gaagacgagg ccctgggtgg
     4201 cacggctttt gttcagatgc cggcccacgg cctattcccc tggtgcggcc tgctgctgga
     4261 tacccggacc ctggaggtgc agagcgacta ctccagctat gcccggacct ccatcagagc
     4321 cagtctcacc ttcaaccgcg gcttcaaggc tgggaggaac atgcgtcgca aactctttgg
     4381 ggtcttgcgg ctgaagtgtc acagcctgtt tctggatttg caggtgaaca gcctccagac
     4441 ggtgtgcacc aacatctaca agatcctcct gctgcaggcg tacaggtttc acgcatgtgt
     4501 gctgcagctc ccatttcatc agcaagtttg gaagaacccc acatttttcc tgcgcgtcat
     4561 ctctgacacg gcctccctct gctactccat cctgaaagcc aagaacgcag ggatgtcgct
     4621 gggggccaag ggcgccgccg gccctctgcc ctccgaggcc gtgcagtggc tgtgccacca
     4681 agcattcctg ctcaagctga ctcgacaccg tgtcacctac gtgccactcc tggggtcact
     4741 caggacagcc cagacgcagc tgagtcggaa gctcccgggg acgacgctga ctgccctgga
     4801 ggccgcagcc aacccggcac tgccctcaga cttcaagacc atcctggact gagtcgaccc
     4861 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4921 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4981 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     5041 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5101 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     5161 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttaccatg accgagtaca
     5221 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     5281 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     5341 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     5401 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     5461 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     5521 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     5581 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     5641 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     5701 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     5761 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg
     5821 acccgcagcg cccgaccgaa aggagcgcac gaccccatgc atcgataaaa taaaagattt
     5881 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     5941 cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata gagaagttca
     6001 gatcaaggtc aggaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
     6061 gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat gggccaaaca
     6121 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
     6181 tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
     6241 acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt
     6301 cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact cggggcgcca
     6361 gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg
     6421 catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg
     6481 tcagcggggg tctttcacat gcagcatgta tcaaaattaa tttggttttt tttcttaagt
     6541 atttacatta aatggccata gttgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6601 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6661 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6721 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6781 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6841 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6901 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6961 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     7021 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7081 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7141 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7201 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     7261 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7321 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7381 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7441 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttgcgg
     7501 ccgcaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7561 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7621 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7681 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7741 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7801 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7861 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7921 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7981 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8041 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8101 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8161 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8221 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8281 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8341 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8401 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8461 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacattt
//
