LOCUS Exported 8709 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Retroviral expression of hTERT. Used to create immortalized cell lines.. ACCESSION . VERSION . KEYWORDS pBABE-neo-hTERT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8709) 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 8709) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8709 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 11..480 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 545..744 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 945..1361 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1291..1313 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1329..1345 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" CDS 1441..4839 /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(4851..4871) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4856..5185 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5036..5171 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5098..5117 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5542..6336 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5596..5615) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6206..6225 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 6859..6878 /label=pBR322ori-F /note="pBR322 origin, forward primer" LTR 7140..7609 /note="long terminal repeat from Moloney murine leukemia virus" CDS complement(7724..8584) /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 8347..8366 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8585..8689) /gene="bla" /label=AmpR promoter ORIGIN 1 cacctgcagc ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc 61 tcagggccaa gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc 121 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag 181 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt 241 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc 301 cgagctcaat aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga 361 gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct 421 cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca 481 tttgggggct cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg 541 aggcaagctg gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt 601 ttatgcgcct gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg 661 gaactgacga gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg 721 gccgtttttg tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat 781 atgtggttct ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc 841 tttcggtttg gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt 901 gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca 961 ctcccttaag tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt 1021 cggtagatgt caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta 1081 acgtcggatg gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca 1141 aggtcttttc acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg 1201 aagccttggc ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc 1261 ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc 1321 gatcctccct ttatccagcc ctcactcctt ctctaggcgc cggccggatc ccagtgtggt 1381 ggtacgtagg aattcgtcct gctgcgcacg tgggaagccc tggccccggc cacccccgcg 1441 atgccgcgcg ctccccgctg ccgagccgtg cgctccctgc tgcgcagcca ctaccgcgag 1501 gtgctgccgc tggccacgtt cgtgcggcgc ctggggcccc agggctggcg gctggtgcag 1561 cgcggggacc cggcggcttt ccgcgcgctg gtggcccagt gcctggtgtg cgtgccctgg 1621 gacgcacggc cgccccccgc cgccccctcc ttccgccagg tgtcctgcct gaaggagctg 1681 gtggcccgag tgctgcagag gctgtgcgag cgcggcgcga agaacgtgct ggccttcggc 1741 ttcgcgctgc tggacggggc ccgcgggggc ccccccgagg ccttcaccac cagcgtgcgc 1801 agctacctgc ccaacacggt gaccgacgca ctgcggggga gcggggcgtg ggggctgctg 1861 ctgcgccgcg tgggcgacga cgtgctggtt cacctgctgg cacgctgcgc gctctttgtg 1921 ctggtggctc ccagctgcgc ctaccaggtg tgcgggccgc cgctgtacca gctcggcgct 1981 gccactcagg cccggccccc gccacacgct agtggacccc gaaggcgtct gggatgcgaa 2041 cgggcctgga accatagcgt cagggaggcc ggggtccccc tgggcctgcc agccccgggt 2101 gcgaggaggc gcgggggcag tgccagccga agtctgccgt tgcccaagag gcccaggcgt 2161 ggcgctgccc ctgagccgga gcggacgccc gttgggcagg ggtcctgggc ccacccgggc 2221 aggacgcgtg gaccgagtga ccgtggtttc tgtgtggtgt cacctgccag acccgccgaa 2281 gaagccacct ctttggaggg tgcgctctct ggcacgcgcc actcccaccc atccgtgggc 2341 cgccagcacc acgcgggccc cccatccaca tcgcggccac cacgtccctg ggacacgcct 2401 tgtcccccgg tgtacgccga gaccaagcac ttcctctact cctcaggcga caaggagcag 2461 ctgcggccct ccttcctact cagctctctg aggcccagcc tgactggcgc tcggaggctc 2521 gtggagacca tctttctggg ttccaggccc tggatgccag ggactccccg caggttgccc 2581 cgcctgcccc agcgctactg gcaaatgcgg cccctgtttc tggagctgct tgggaaccac 2641 gcgcagtgcc cctacggggt gctcctcaag acgcactgcc cgctgcgagc tgcggtcacc 2701 ccagcagccg gtgtctgtgc ccgggagaag ccccagggct ctgtggcggc ccccgaggag 2761 gaggacacag acccccgtcg cctggtgcag ctgctccgcc agcacagcag cccctggcag 2821 gtgtacggct tcgtgcgggc ctgcctgcgc cggctggtgc ccccaggcct ctggggctcc 2881 aggcacaacg aacgccgctt cctcaggaac accaagaagt tcatctccct ggggaagcat 2941 gccaagctct cgctgcagga gctgacgtgg aagatgagcg tgcggggctg cgcttggctg 3001 cgcaggagcc caggggttgg ctgtgttccg gccgcagagc accgtctgcg tgaggagatc 3061 ctggccaagt tcctgcactg gctgatgagt gtgtacgtcg tcgagctgct caggtctttc 3121 ttttatgtca cggagaccac gtttcaaaag aacaggctct ttttctaccg gaagagtgtc 3181 tggagcaagt tgcaaagcat tggaatcaga cagcacttga agagggtgca gctgcgggag 3241 ctgtcggaag cagaggtcag gcagcatcgg gaagccaggc ccgccctgct gacgtccaga 3301 ctccgcttca tccccaagcc tgacgggctg cggccgattg tgaacatgga ctacgtcgtg 3361 ggagccagaa cgttccgcag agaaaagagg gccgagcgtc tcacctcgag ggtgaaggca 3421 ctgttcagcg tgctcaacta cgagcgggcg cggcgccccg gcctcctggg cgcctctgtg 3481 ctgggcctgg acgatatcca cagggcctgg cgcaccttcg tgctgcgtgt gcgggcccag 3541 gacccgccgc ctgagctgta ctttgtcaag gtggatgtga cgggcgcgta cgacaccatc 3601 ccccaggaca ggctcacgga ggtcatcgcc agcatcatca aaccccagaa cacgtactgc 3661 gtgcgtcggt atgccgtggt ccagaaggcc gcccatgggc acgtccgcaa ggccttcaag 3721 agccacgtct ctaccttgac agacctccag ccgtacatgc gacagttcgt ggctcacctg 3781 caggagacca gcccgctgag ggatgccgtc gtcatcgagc agagctcctc cctgaatgag 3841 gccagcagtg gcctcttcga cgtcttccta cgcttcatgt gccaccacgc cgtgcgcatc 3901 aggggcaagt cctacgtcca gtgccagggg atcccgcagg gctccatcct ctccacgctg 3961 ctctgcagcc tgtgctacgg cgacatggag aacaagctgt ttgcggggat tcggcgggac 4021 gggctgctcc tgcgtttggt ggatgatttc ttgttggtga cacctcacct cacccacgcg 4081 aaaaccttcc tcaggaccct ggtccgaggt gtccctgagt atggctgcgt ggtgaacttg 4141 cggaagacag tggtgaactt ccctgtagaa gacgaggccc tgggtggcac ggcttttgtt 4201 cagatgccgg cccacggcct attcccctgg tgcggcctgc tgctggatac ccggaccctg 4261 gaggtgcaga gcgactactc cagctatgcc cggacctcca tcagagccag tctcaccttc 4321 aaccgcggct tcaaggctgg gaggaacatg cgtcgcaaac tctttggggt cttgcggctg 4381 aagtgtcaca gcctgtttct ggatttgcag gtgaacagcc tccagacggt gtgcaccaac 4441 atctacaaga tcctcctgct gcaggcgtac aggtttcacg catgtgtgct gcagctccca 4501 tttcatcagc aagtttggaa gaaccccaca tttttcctgc gcgtcatctc tgacacggcc 4561 tccctctgct actccatcct gaaagccaag aacgcaggga tgtcgctggg ggccaagggc 4621 gccgccggcc ctctgccctc cgaggccgtg cagtggctgt gccaccaagc attcctgctc 4681 aagctgactc gacaccgtgt cacctacgtg ccactcctgg ggtcactcag gacagcccag 4741 acgcagctga gtcggaagct cccggggacg acgctgactg ccctggaggc cgcagccaac 4801 ccggcactgc cctcagactt caagaccatc ctggactgag tcgaccctgt ggaatgtgtg 4861 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 4921 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 4981 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc 5041 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat 5101 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt 5161 ttttggaggc ctaggctttt gcaaaaagct tcacgctgcc gcaagcactc agggcgcaag 5221 ggctgctaaa ggaagcggaa cacgtagaaa gccagtccgc agaaacggtg ctgaccccgg 5281 atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa gagaaagcag 5341 gtagcttgca gtgggcttac atggcgatag ctagactggg cggttttatg gacagcaagc 5401 gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac 5461 tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagatc tgatcaagag 5521 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc 5581 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat 5641 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg 5701 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg 5761 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta 5821 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta 5881 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc 5941 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc 6001 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg 6061 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg 6121 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt 6181 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc 6241 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc 6301 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcatcga 6361 tgcatcgatg cggcgggtgt ggtggttacg cgtaatctgc tgcttgcaaa caaaaaaacc 6421 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt 6481 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg 6541 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc 6601 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 6661 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga 6721 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct 6781 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg 6841 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca 6901 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 6961 cgccagcaac gcatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat 7021 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg 7081 gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc 7141 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag 7201 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc 7261 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga 7321 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga 7381 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata 7441 aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt 7501 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt 7561 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac atgcagcatg 7621 tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca tagtgcggcc 7681 gcaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 7741 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 7801 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 7861 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 7921 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 7981 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 8041 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 8101 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 8161 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 8221 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 8281 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 8341 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 8401 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 8461 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 8521 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 8581 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 8641 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 8701 gaaaagtgc //