LOCUS Exported 6694 bp ds-DNA circular SYN 25-MAR-2022 DEFINITION This plasmid is in the pT3-EF1a vector without loxP sites flanking the inverted repeats of SB (sleeping beauty) sequence. Therefore this plasmid can be used with Cre for in vivo studies.. ACCESSION . VERSION . KEYWORDS pT3EF1aH-myr-Akt SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6694) AUTHORS Ho C, Wang C, Mattu S, Destefanis G, Ladu S, Delogu S, Armbruster J, Fan L, Lee SA, Jiang L, Dombrowski F, Evert M, Chen X, Calvisi DF TITLE AKT (v-akt murine thymoma viral oncogene homolog 1) and N-Ras (neuroblastoma ras viral oncogene homolog) coactivation in the mouse liver promotes rapid carcinogenesis by way of mTOR (mammalian target of rapamycin complex 1), FOXM1 (forkhead box M1)/SKP2, and c-Myc pathways. JOURNAL Hepatology. 2012 Mar;55(3):833-45. doi: 10.1002/hep.24736. Epub 2011 Dec 19. PUBMED 21993994 REFERENCE 2 (bases 1 to 6694) AUTHORS . TITLE Direct Submission JOURNAL Exported Mar 25, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6694 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(35..52) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(206..794) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(286..305) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(965..1825) /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 1588..1607 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1826..1930) /gene="bla" /label=AmpR promoter primer_bind 1998..2016 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(2054..2076) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 2176..2195 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2389..2411 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2403..2420 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2404..2420 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2802..2822) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2944..4122 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 3175..4113 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 4070..4090 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" protein_bind 4153..4177 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 4212..4232 /codon_start=1 /product="N-myristoylation signal from Src kinase (Pellman et al., 1985; Kaplan et al., 1988)" /label=myr /translation="MGSSKSK" CDS 5685..5711 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" protein_bind complement(5744..5768) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind 5796..5815 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5850..5867) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5856..6080 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(6498..6514) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6498..6514) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6511..6533) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6522..6538 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(6546..6576) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6591..6612 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 61 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 121 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 721 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 841 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 901 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 961 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 1021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 1081 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 1141 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 1201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 1261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 1321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 1381 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 1441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 1501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 1561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 1621 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 1681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 1741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 1801 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 1861 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 1921 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca 1981 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg 2041 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag 2101 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg 2161 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg 2221 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc 2281 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga 2341 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac 2401 gttgtaaaac gacggccagt gaattcgagc tcggtaccct acagttgaag tcggaagttt 2461 acatacactt aagttggagt cattaaaact cgtttttcaa ctactccaca aatttcttgt 2521 taacaaacaa tagttttggc aagtcagtta ggacatctac tttgtgcatg acacaagtca 2581 tttttccaac aattgtttac agacagatta tttcacttat aattcactgt atcacaattc 2641 cagtgggtca gaagtttaca tacactaagt tgactgtgcc tttaaacagc ttggaaaatt 2701 ccagaaaatg atgtcatggc tttagaagct tctgatagac taattgacat catttgagtc 2761 aattggaggt gtacctgtgg atgtatttca aggaattctg tggaatgtgt gtcagttagg 2821 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atatcgatac 2881 tagtttaatt aagatcctct ttgcagctaa tggaccttct aggtcttgaa aggagtggga 2941 attggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg 3001 gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa 3061 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt 3121 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt 3181 gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga 3241 attacttcca cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg 3301 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc 3361 tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct 3421 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc 3481 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg 3541 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg 3601 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg 3661 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca 3721 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg 3781 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt 3841 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc 3901 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg 3961 atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg 4021 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag 4081 acagtggttc aaagtttttt tcttccattt caggtgtcgt gaggaattag cttggtacca 4141 atacggatat caacaagttt gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgac 4201 tggatcccac catggggagc agcaagagca agcccaagtc tagaatgaac gacgtagcca 4261 ttgtgaagga gggctggctg cacaaacgag gggaatatat taaaacctgg cggccacgct 4321 acttcctcct caagaacgat ggcaccttta ttggctacaa ggaacggcct caggatgtgg 4381 atcagcgaga gtccccactc aacaacttct cagtggcaca atgccagctg atgaagacag 4441 agcggccaag gcccaacacc tttatcatcc gctgcctgca gtggaccaca gtcattgagc 4501 gcaccttcca tgtggaaacg cctgaggagc gggaagaatg ggccaccgcc attcagactg 4561 tggccgatgg actcaagagg caggaagaag agacgatgga cttccgatca ggctcaccca 4621 gtgacaactc aggggctgaa gagatggagg tgtccctggc caagcccaag caccgtgtga 4681 ccatgaacga gtttgagtac ctgaaactac tgggcaaggg cacctttggg aaagtgattc 4741 tggtgaaaga gaaggccaca ggccgctact atgccatgaa gatcctcaag aaggaggtca 4801 tcgtcgccaa ggatgaggtt gcccacacgc ttactgagaa ccgtgtcctg cagaactcta 4861 ggcatccctt ccttacggcc ctcaagtact cattccagac ccacgaccgc ctctgctttg 4921 tcatggagta tgccaacggg ggcgagctct tcttccacct gtctcgagag cgcgtgttct 4981 ccgaggaccg ggcccgcttc tatggtgcgg agattgtgtc tgccctggac tacttgcact 5041 ccgagaagaa cgtggtgtac cgggacctga agctggagaa cctcatgctg gacaaggacg 5101 ggcacatcaa gataacggac ttcgggctgt gcaaggaggg gatcaaggat ggtgccacta 5161 tgaagacatt ctgcggaacg ccggagtacc tggcccctga ggtgctggag gacaacgact 5221 acggccgtgc agtggactgg tgggggctgg gcgtggtcat gtatgagatg atgtgtggcc 5281 gcctgccctt ctacaaccag gaccacgaga agctgttcga gctgatcctc atggaggaga 5341 tccgcttccc gcgcacactc ggccctgagg ccaagtccct gctctccggg ctgctcaaga 5401 aggaccctac acagaggctc ggtgggggct ctgaggatgc caaggagatc atgcagcacc 5461 ggttctttgc caacatcgtg tggcaggatg tgtatgagaa gaagctgagc ccacctttca 5521 agccccaggt cacctctgag actgacacca ggtatttcga tgaggagttc acagctcaga 5581 tgatcaccat cacgccgcct gatcaagatg acagcatgga gtgtgtggac agtgagcgga 5641 ggccgcactt cccccagttc tcctactcag ccagtggcac agcctaccca tacgacgtcc 5701 cagactacgc ttgagaattc gcggccgcac tcgagatatc tagacccagc tttcttgtac 5761 aaagtggttg atatcgcggc cgctgcattc tagttgtggt ttgtccaaac tcatcaatgt 5821 atcttatcat gtctggatcc gctgatcagc ctcgactgtg ccttctagtt gccagccatc 5881 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct 5941 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg 6001 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg 6061 ggatgcggtg ggctctatgg cttctgaggc gggatcccct tgaaatacat ccacaggtac 6121 acctccaatt gactcaaatg atgtcaatta gtctatcaga agcttctaaa gccatgacat 6181 cattttctgg aattttccaa gctgtttaaa ggcacagtca acttagtgta tgtaaacttc 6241 tgacccactg gaattgtgat acagtgaatt ataagtgaaa taatctgtct gtaaacaatt 6301 gttggaaaaa tgacttgtgt catgcacaaa gtagatgtcc taactgactt gccaaaacta 6361 ttgtttgtta acaagaaatt tgtggagtag ttgaaaaacg agttttaatg actccaactt 6421 aagtgtatgt aaacttccga cttcaactgt ataggtctag agtcgacctg caggcatgca 6481 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 6541 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 6601 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 6661 cagctgcatt aatgaatcgg ccaacgcgcg ggga //