LOCUS Exported 6494 bp ds-DNA circular SYN 14-FEB-2022 DEFINITION Expresses c-Myc in mammalian cells. ACCESSION . VERSION . KEYWORDS c-myc-PT3EF1a SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6494) AUTHORS Liu P, Ge M, Hu J, Li X, Che L, Sun K, Cheng L, Huang Y, Pilo MG, Cigliano A, Pes GM, Pascale RM, Brozzetti S, Vidili G, Porcu A, Cossu A, Palmieri G, Sini MC, Ribback S, Dombrowski F, Tao J, Calvisi DF, Chen L, Chen X TITLE A functional mTORC1 signaling is indispensable for c-Myc driven hepatocarcinogenesis. JOURNAL Hepatology. 2017 Mar 30. doi: 10.1002/hep.29183. PUBMED 28370287 REFERENCE 2 (bases 1 to 6494) AUTHORS . TITLE Direct Submission JOURNAL Exported Feb 14, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6494 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 133..152 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 386..403 /label=L4440 /note="L4440 vector, forward primer" protein_bind 520..541 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 556..586 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 594..610 /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)." primer_bind 599..621 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 618..634 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 618..634 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" polyA_signal 1052..1276 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind 1265..1282 /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" primer_bind complement(1317..1336) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" protein_bind 1364..1388 /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS complement(1405..2721) /codon_start=1 /product="human c-Myc proto-oncogene" /label=c-Myc /translation="MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAP SEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTE LLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPN PARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDS LLSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGS PSAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNR KCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKA TAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSCA" protein_bind complement(2755..2779) /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" promoter complement(2810..3988) /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2819..3757 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind complement(2842..2862) /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" primer_bind 4110..4130 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" primer_bind complement(4512..4529) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4512..4528) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4521..4543) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(4737..4756) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4856..4878 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4916..4934) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5002..5106 /gene="bla" /label=AmpR promoter CDS 5107..5967 /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 complement(5325..5344) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6138..232 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga 61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag 121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg 181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag 241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc 301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc 361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc 421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag 481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca 541 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag 601 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgcatgcct 661 gcaggtcgac tctagaccta tacagttgaa gtcggaagtt tacatacact taagttggag 721 tcattaaaac tcgtttttca actactccac aaatttcttg ttaacaaaca atagttttgg 781 caagtcagtt aggacatcta ctttgtgcat gacacaagtc atttttccaa caattgttta 841 cagacagatt atttcactta taattcactg tatcacaatt ccagtgggtc agaagtttac 901 atacactaag ttgactgtgc ctttaaacag cttggaaaat tccagaaaat gatgtcatgg 961 ctttagaagc ttctgataga ctaattgaca tcatttgagt caattggagg tgtacctgtg 1021 gatgtatttc aaggggatcc cgcctcagaa gccatagagc ccaccgcatc cccagcatgc 1081 ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc ccccagaata 1141 gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga aaggacagtg 1201 ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac agatggctgg 1261 caactagaag gcacagtcga ggctgatcag cggatccaga catgataaga tacattgatg 1321 agtttggaca aaccacaact agaatgcagc ggccgcgata tcaaccactt tgtacaagaa 1381 agctgggtct agatatctcg agtccgcaca agagttccgt agctgttcaa gtttgtgttt 1441 caactgttct cgtcgtttcc gcaacaagtc ctcttcagaa atgagctttt gctcctctgc 1501 ttggacggac aggatgtatg ctgtggcttt tttaaggata actaccttgg gggccttttc 1561 attgttttcc aactccggga tctggtcacg cagggcaaaa aagctccgtt ttagctcgtt 1621 cctcctctgg cgctccaaga cgttgtgtgt tcgcctcttg acattctcct cggtgtccga 1681 ggacctgggg ctggtgcatt ttcggttgtt gctgatctgt ctcaggactc tgacactgtc 1741 caacttgacc ctcttggcag caggatagtc cttccgagtg gagggaggcg ctgcgtagtt 1801 gtgctgatgt gtggagacgt ggcacctctt gaggaccagt gggctgtgag gaggtttgct 1861 gtggcctcca gcagaaggtg atccagactc tgaccttttg ccaggagcct gcctcttttc 1921 cacagaaaca acatcgattt cttcctcatc ttcttgttcc tcctcagagt cgctgctggt 1981 ggtgggcggt gtctcctcat ggagcaccag gggctcgggg ctgccctgcg gggaggactc 2041 cgtcgaggag agcagagaat ccgaggacgg agagaaggcg ctggagtctt gcgaggcgca 2101 ggacttgggc gagctgctgt cgttgagagg gtaggggaag accaccgagg ggtcgatgca 2161 ctctgaggcg gcggcgctca gatcctgcag gtacaagctg gaggtggagc agacgctgtg 2221 gccgcgggcg gggttcgggc tgccgctgtc tttgcgcgca gcctggtagg aggccagctt 2281 ctctgagacg agcttggcgg cggccgagaa gccgctccac atacagtcct ggatgatgat 2341 gtttttgatg aaggtctcgt cgtccgggtc gcagatgaaa ctctggttca ccatgtctcc 2401 tcccagcagc tcggtcacca tctccagctg gtcggccgtg gagaagctcc cgccaccgcc 2461 gtcgttgtct ccccgaaggg agaagggtgt gaccgcaacg taggagggcg agcagagccc 2521 ggagcggcgg ctaggggaca ggggcggggt gggcagcagc tcgaatttct tccagatatc 2581 ctcgctgggc gccgggggct gcagctcgct ctgctgctgc tgctggtaga agttctcctc 2641 ctcgtcgcag tagaaatacg gctgcaccga gtcgtagtcg aggtcatagt tcctgttggt 2701 gaagctaacg ttgaggggca tggtggatcc agtcgactga attggttcct ttaaagcctg 2761 cttttttgta caaacttgtt gatatccgta ttggtaccaa gctaattcct cacgacacct 2821 gaaatggaag aaaaaaactt tgaaccactg tctgaggctt gagaatgaac caagatccaa 2881 actcaaaaag ggcaaattcc aaggagaatt acatcaagtg ccaagctggc ctaacttcag 2941 tctccaccca ctcagtgtgg ggaaactcca tcgcataaaa cccctccccc caacctaaag 3001 acgacgtact ccaaaagctc gagaactaat cgaggtgcct ggacggcgcc cggtactccg 3061 tggagtcaca tgaagcgacg gctgaggacg gaaaggccct tttcctttgt gtgggtgact 3121 cacccgcccg ctctcccgag cgccgcgtcc tccattttga gctccctgca gcagggccgg 3181 gaagcggcca tctttccgct cacgcaactg gtgccgaccg ggccagcctt gccgcccagg 3241 gcggggcgat acacggcggc gcgaggccag gcaccagagc aggccggcca gcttgagact 3301 acccccgtcc gattctcggt ggccgcgctc gcaggccccg cctcgccgaa catgtgcgct 3361 gggacgcacg ggccccgtcg ccgcccgcgg ccccaaaaac cgaaatacca gtgtgcagat 3421 cttggcccgc atttacaaga ctatcttgcc agaaaaaaag cgtcgcagca ggtcatcaaa 3481 aattttaaat ggctagagac ttatcgaaag cagcgagaca ggcgcgaagg tgccaccaga 3541 ttcgcacgcg gcggccccag cgcccaggcc aggcctcaac tcaagcacga ggcgaagggg 3601 ctccttaagc gcaaggcctc gaactctccc acccacttcc aacccgaagc tcgggatcaa 3661 gaatcacgta ctgcagccag gtggaagtaa ttcaaggcac gcaagggcca taacccgtaa 3721 agaggccagg cccgcgggaa ccacacacgg cacttacctg tgttctggcg gcaaacccgt 3781 tgcgaaaaag aacgttcacg gcgactactg cacttatata cggttctccc ccaccctcgg 3841 gaaaaaggcg gagccagtac acgacatcac tttcccagtt taccccgcgc caccttctct 3901 aggcaccggt tcaattgccg acccctcccc ccaacttctc ggggactgtg ggcgatgtgc 3961 gctctgccca ctgacgggca ccggagccaa ttcccactcc tttcaagacc tagaaggtcc 4021 attagctgca aagaggatct taattaaact agtatcgata tgcatgcttt gcatacttct 4081 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagaattcc 4141 ttgaaataca tccacaggta cacctccaat tgactcaaat gatgtcaatt agtctatcag 4201 aagcttctaa agccatgaca tcattttctg gaattttcca agctgtttaa aggcacagtc 4261 aacttagtgt atgtaaactt ctgacccact ggaattgtga tacagtgaat tataagtgaa 4321 ataatctgtc tgtaaacaat tgttggaaaa atgacttgtg tcatgcacaa agtagatgtc 4381 ctaactgact tgccaaaact attgtttgtt aacaagaaat ttgtggagta gttgaaaaac 4441 gagttttaat gactccaact taagtgtatg taaacttccg acttcaactg tagggtaccg 4501 agctcgaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc 4561 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc 4621 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg 4681 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca 4741 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg 4801 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg 4861 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc 4921 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt 4981 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca 5041 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg 5101 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc 5161 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg 5221 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt 5281 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta 5341 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat 5401 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga 5461 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca 5521 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact 5581 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc 5641 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact 5701 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt 5761 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt 5821 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt 5881 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata 5941 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag 6001 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat 6061 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa 6121 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 6181 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 6241 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg 6301 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 6361 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 6421 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 6481 agcttggagc gaac 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