LOCUS       sequence-357764-        6694 bp    DNA     circular SYN 15-DEC-2025
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    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1002/hep.24736";
            journalName: "Hepatology"; date: "2012-03"; volume: "55"; issue: 
            "3"; pages: "833-45"
COMMENT     Sequence Label: pT3EF1aH-myr-Akt
FEATURES             Location/Qualifiers
     source          1..6694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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(join(965..1756,1757..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(join(6546..6552,6553..6570,6571..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
//
