LOCUS Exported 8651 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Retroviral expression of hTERT. Used to create immortalized cell lines.. ACCESSION . VERSION . KEYWORDS pBABE-hygro-hTERT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8651) 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 8651) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8651 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 7..476 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 541..740 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 941..1357 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1287..1309 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1325..1341 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" CDS 1437..4835 /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(4847..4867) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4852..5181 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5032..5167 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5094..5113 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5255..6280 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" primer_bind 6797..6816 /label=pBR322ori-F /note="pBR322 origin, forward primer" LTR 7078..7547 /note="long terminal repeat from Moloney murine leukemia virus" CDS complement(7662..8522) /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 8285..8304 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8523..8627) /gene="bla" /label=AmpR promoter ORIGIN 1 tgcagcctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag 61 ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt 121 cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc cctcagcagt 181 ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct 241 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag 301 ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt gactgagtcg 361 cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct 421 gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatttg 481 ggggctcgtc cgggatcggg agacccctgc ccagggacca ccgacccacc accgggaggc 541 aagctggcca gcaacttatc tgtgtctgtc cgattgtcta gtgtctatga ctgattttat 601 gcgcctgcgt cggtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac 661 tgacgagttc tgaacacccg gccgcaaccc tgggagacgt cccagggact ttgggggccg 721 tttttgtggc ccgacctgag gaagggagtc gatgtggaat ccgaccccgt caggatatgt 781 ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc 841 ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg 901 tctctgtctg actgtgtttc tgtatttgtc tgaaaattag ggccagactg ttaccactcc 961 cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt 1021 agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt 1081 cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt 1141 cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc 1201 cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc ctccgcctcc 1261 tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc cgcctcgatc 1321 ctccctttat ccagccctca ctccttctct aggcgccggc cggatcccag tgtggtggta 1381 cgtaggaatt cgtcctgctg cgcacgtggg aagccctggc cccggccacc cccgcgatgc 1441 cgcgcgctcc ccgctgccga gccgtgcgct ccctgctgcg cagccactac cgcgaggtgc 1501 tgccgctggc cacgttcgtg cggcgcctgg ggccccaggg ctggcggctg gtgcagcgcg 1561 gggacccggc ggctttccgc gcgctggtgg cccagtgcct ggtgtgcgtg ccctgggacg 1621 cacggccgcc ccccgccgcc ccctccttcc gccaggtgtc ctgcctgaag gagctggtgg 1681 cccgagtgct gcagaggctg tgcgagcgcg gcgcgaagaa cgtgctggcc ttcggcttcg 1741 cgctgctgga cggggcccgc gggggccccc ccgaggcctt caccaccagc gtgcgcagct 1801 acctgcccaa cacggtgacc gacgcactgc gggggagcgg ggcgtggggg ctgctgctgc 1861 gccgcgtggg cgacgacgtg ctggttcacc tgctggcacg ctgcgcgctc tttgtgctgg 1921 tggctcccag ctgcgcctac caggtgtgcg ggccgccgct gtaccagctc ggcgctgcca 1981 ctcaggcccg gcccccgcca cacgctagtg gaccccgaag gcgtctggga tgcgaacggg 2041 cctggaacca tagcgtcagg gaggccgggg tccccctggg cctgccagcc ccgggtgcga 2101 ggaggcgcgg gggcagtgcc agccgaagtc tgccgttgcc caagaggccc aggcgtggcg 2161 ctgcccctga gccggagcgg acgcccgttg ggcaggggtc ctgggcccac ccgggcagga 2221 cgcgtggacc gagtgaccgt ggtttctgtg tggtgtcacc tgccagaccc gccgaagaag 2281 ccacctcttt ggagggtgcg ctctctggca cgcgccactc ccacccatcc gtgggccgcc 2341 agcaccacgc gggcccccca tccacatcgc ggccaccacg tccctgggac acgccttgtc 2401 ccccggtgta cgccgagacc aagcacttcc tctactcctc aggcgacaag gagcagctgc 2461 ggccctcctt cctactcagc tctctgaggc ccagcctgac tggcgctcgg aggctcgtgg 2521 agaccatctt tctgggttcc aggccctgga tgccagggac tccccgcagg ttgccccgcc 2581 tgccccagcg ctactggcaa atgcggcccc tgtttctgga gctgcttggg aaccacgcgc 2641 agtgccccta cggggtgctc ctcaagacgc actgcccgct gcgagctgcg gtcaccccag 2701 cagccggtgt ctgtgcccgg gagaagcccc agggctctgt ggcggccccc gaggaggagg 2761 acacagaccc ccgtcgcctg gtgcagctgc tccgccagca cagcagcccc tggcaggtgt 2821 acggcttcgt gcgggcctgc ctgcgccggc tggtgccccc aggcctctgg ggctccaggc 2881 acaacgaacg ccgcttcctc aggaacacca agaagttcat ctccctgggg aagcatgcca 2941 agctctcgct gcaggagctg acgtggaaga tgagcgtgcg gggctgcgct tggctgcgca 3001 ggagcccagg ggttggctgt gttccggccg cagagcaccg tctgcgtgag gagatcctgg 3061 ccaagttcct gcactggctg atgagtgtgt acgtcgtcga gctgctcagg tctttctttt 3121 atgtcacgga gaccacgttt caaaagaaca ggctcttttt ctaccggaag agtgtctgga 3181 gcaagttgca aagcattgga atcagacagc acttgaagag ggtgcagctg cgggagctgt 3241 cggaagcaga ggtcaggcag catcgggaag ccaggcccgc cctgctgacg tccagactcc 3301 gcttcatccc caagcctgac gggctgcggc cgattgtgaa catggactac gtcgtgggag 3361 ccagaacgtt ccgcagagaa aagagggccg agcgtctcac ctcgagggtg aaggcactgt 3421 tcagcgtgct caactacgag cgggcgcggc gccccggcct cctgggcgcc tctgtgctgg 3481 gcctggacga tatccacagg gcctggcgca ccttcgtgct gcgtgtgcgg gcccaggacc 3541 cgccgcctga gctgtacttt gtcaaggtgg atgtgacggg cgcgtacgac accatccccc 3601 aggacaggct cacggaggtc atcgccagca tcatcaaacc ccagaacacg tactgcgtgc 3661 gtcggtatgc cgtggtccag aaggccgccc atgggcacgt ccgcaaggcc ttcaagagcc 3721 acgtctctac cttgacagac ctccagccgt acatgcgaca gttcgtggct cacctgcagg 3781 agaccagccc gctgagggat gccgtcgtca tcgagcagag ctcctccctg aatgaggcca 3841 gcagtggcct cttcgacgtc ttcctacgct tcatgtgcca ccacgccgtg cgcatcaggg 3901 gcaagtccta cgtccagtgc caggggatcc cgcagggctc catcctctcc acgctgctct 3961 gcagcctgtg ctacggcgac atggagaaca agctgtttgc ggggattcgg cgggacgggc 4021 tgctcctgcg tttggtggat gatttcttgt tggtgacacc tcacctcacc cacgcgaaaa 4081 ccttcctcag gaccctggtc cgaggtgtcc ctgagtatgg ctgcgtggtg aacttgcgga 4141 agacagtggt gaacttccct gtagaagacg aggccctggg tggcacggct tttgttcaga 4201 tgccggccca cggcctattc ccctggtgcg gcctgctgct ggatacccgg accctggagg 4261 tgcagagcga ctactccagc tatgcccgga cctccatcag agccagtctc accttcaacc 4321 gcggcttcaa ggctgggagg aacatgcgtc gcaaactctt tggggtcttg cggctgaagt 4381 gtcacagcct gtttctggat ttgcaggtga acagcctcca gacggtgtgc accaacatct 4441 acaagatcct cctgctgcag gcgtacaggt ttcacgcatg tgtgctgcag ctcccatttc 4501 atcagcaagt ttggaagaac cccacatttt tcctgcgcgt catctctgac acggcctccc 4561 tctgctactc catcctgaaa gccaagaacg cagggatgtc gctgggggcc aagggcgccg 4621 ccggccctct gccctccgag gccgtgcagt ggctgtgcca ccaagcattc ctgctcaagc 4681 tgactcgaca ccgtgtcacc tacgtgccac tcctggggtc actcaggaca gcccagacgc 4741 agctgagtcg gaagctcccg gggacgacgc tgactgccct ggaggccgca gccaacccgg 4801 cactgccctc agacttcaag accatcctgg actgagtcga ccctgtggaa tgtgtgtcag 4861 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 4921 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 4981 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc 5041 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat 5101 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt 5161 ggaggcctag gcttttgcaa aaagcttgcc agctggggcg ccctctggta aggttgggaa 5221 gccctgcaga tctgatcccg gggggcaatg agatatgaaa aagcctgaac tcaccgcgac 5281 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc 5341 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg 5401 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc 5461 ggccgcgctc ccgattccgg aagtgcttga cattggggaa tttagcgaga gcctgaccta 5521 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc 5581 cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca 5641 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga 5701 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac 5761 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc 5821 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg 5881 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt 5941 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt 6001 cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat 6061 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc 6121 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat 6181 cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg 6241 aaaccgacgc cccagcactc gtccgagggc aaaggaatag agtagatgcc gaccgggatc 6301 tatcgatgcg gcgggtgtgg tggttacgcg taatctgctg cttgcaaaca aaaaaaccac 6361 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 6421 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc 6481 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 6541 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 6601 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 6661 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc 6721 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 6781 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 6841 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 6901 ccagcaacgc atcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga 6961 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga 7021 aaaatacata actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagctg 7081 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa 7141 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg 7201 gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga 7261 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac 7321 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa 7381 agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac 7441 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg 7501 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcacat gcagcatgta 7561 tcaaaattaa tttggttttt tttcttaagt atttacatta aatggccata gtgcggccgc 7621 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 7681 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 7741 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 7801 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 7861 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 7921 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 7981 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 8041 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 8101 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 8161 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 8221 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 8281 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 8341 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 8401 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 8461 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 8521 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 8581 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 8641 aaagtgccac c //