LOCUS Exported 7151 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS GSK3 alpha pMT2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7151) TITLE GSK3 plasmids REFERENCE 2 (bases 1 to 7151) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7151 /organism="synthetic DNA construct" /mol_type="other DNA" promoter complement(7..336) /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 21..156 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(75..94) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 321..341 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" intron 805..1034 /label=chimeric intron /note="chimera between introns from adenovirus and immunoglobulin heavy chain genes" primer_bind 1017..1036 /label=pMT2-F /note="Synthetic intron, forward primer" CDS 3351..3914 /codon_start=1 /gene="Mus musculus Dhfr" /product="mouse dihydrofolate reductase" /label=DHFR /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY PGVLSEVQEEKGIKYKFEVYEKKD" polyA_signal 4102..4236 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4139..4158) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4193..4212 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4763..4780) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4934..5522) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5014..5033) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5693..6553) /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 6316..6335 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6554..6658) /gene="bla" /label=AmpR promoter primer_bind 6726..6744 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(6782..6804) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 6904..6923 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7117..7139 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 7131..7148 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 7132..7148 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 aagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc 61 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg 121 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat 181 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga 241 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg 301 ggagcctggg gactttccac accctaactg acacacattc cacaggatcc ggtcgcgcga 361 atttcgagcg gtgttccgcg gtcctcctcg tatagaaact cggaccactc tgagacgaag 421 gctcgcgtcc aggccagcac gaaggaggct aagtgggagg ggtagcggtc gttgtccact 481 agggggtcca ctcgctccag ggtgtgaaga cacatgtcgc cctcttcggc atcaaggaag 541 gtgattggtt tataggtgta ggccacgtga ccgggtgttc ctgaaggggg gctataaaag 601 ggggtggggg cgcgttcgtc ctcactctct tccgcatcgc tgtctgcgag ggccagctgt 661 tgggctcgcg gttgaggaca aactcttcgc ggtctttcca gtactcttgg atcggaaacc 721 cgtcggcctc cgaacggtac tccgccaccg agggacctga gcgagtccgc atcgaccgga 781 tcggaaaacc tctcgactgt tggggtgagt actccctctc aaaagcgggc atgacttctg 841 cgctaagatt gtcagtttcc aaaaacgagg aggatttgat attcacctgg cccgcggtga 901 tgcctttgag ggtggccgcg tccatctggt cagaaaagac aatctttttg ttgtcaagct 961 tgaggtgtgg caggcttgag atctggccat acacttgagt gacaatgaca tccactttgc 1021 ctttctctcc acaggtgtcc actcccaggt ccaactgcag gcgagcctga attcgccaga 1081 gcggcgcggc ctggaagagg ccagggcccg ggggaggcga cggcagcggc ggcggctggg 1141 gcagcccggg cagcccgagc cccgcagcct gggcctgtgc tcggcgccat gagcggcggc 1201 gggccttcgg gaggcggccc tgggggctcg ggcagggcgc ggactagctc gttcgcggag 1261 cccggcggcg gaggcggagg aggcggcggc ggccccggag gctcggcctc cggcccaggc 1321 ggcaccggcg gcggaaaggc atctgtcggg gccatgggtg ggggcgtcgg ggcctcgagc 1381 tccgggggtg gacccggcgg cagcggcgga ggaggcagcg gaggccccgg cgcaggcact 1441 agcttcccgc cgcccggggt gaagctgggc cgtgacagcg ggaaggtgac cacagtcgta 1501 gccactctag gccaaggccc agagcgctcc caagaagtgg cttacacgga catcaaagtg 1561 attggcaatg gctcatttgg ggtcgtgtac caggcacggc tggcagagac cagggaacta 1621 gtcgccatca agaaggttct ccaggacaag aggttcaaga accgagagct gcagatcatg 1681 cgtaagctgg accactgcaa tattgtgagg ctgagatact ttttctactc cagtggcgag 1741 aagaaagacg agctttacct aaatctggtg ctggaatatg tgcccgagac agtgtaccgg 1801 gtggcccgcc acttcaccaa ggccaagttg accatcccta tcctctatgt caaggtgtac 1861 atgtaccagc tcttccgcag cttggcctac atccactccc agggcgtgtg tcaccgcgac 1921 atcaagcccc agaacctgct ggtggaccct gacactgctg tcctcaagct ctgcgatttt 1981 ggcagtgcaa agcagttggt ccgaggggag cccaatgtct cctacatctg ttctcgctac 2041 taccgggccc cagagctcat ctttggagcc actgattaca cctcatccat cgatgtttgg 2101 tcagctggct gtgtactggc agagctcctc ttgggccagc ccatcttccc tggggacagt 2161 ggggtggacc agctggtgga gatcatcaag gtgctgggaa caccaacccg ggaacaaatc 2221 cgagagatga accccaacta cacggagttc aagttccctc agattaaagc tcacccctgg 2281 acaaaggtgt tcaaatctcg aacgccgcca gaggccatcg cgctctgctc tagcctgctg 2341 gagtacaccc catcctcaag gctctcccca ctagaggcct gtgcgcacag cttctttgat 2401 gaactgcgat gtctgggaac ccagctgcct aacaaccgcc cacttccccc tctcttcaac 2461 ttcagtgctg gtgaactctc catccaaccg tctctcaacg ccattctcat ccctcctcac 2521 ttgaggtccc ccagcggcac taccaccctc accccgtcct cacaagcttt aactgagact 2581 ccgaccagct cagactggca gtcgaccgat gccacaccta ccctcactaa ctcctcctga 2641 gggccccacc aagcaccctt ccacttccat ctgggagccc caagagggcg tgggaagggg 2701 ggccatagcc catcaagctc ctgccctggc tgggccccta gactagaggg cagaggtaaa 2761 tgagtccctg tccccacctc cagtccctcc ctcaccagcc tcacccctgt ggtgggcttt 2821 ttaagaggat tttaactggt tgtggggagg gaagagaagg acagggtgtt ggggggatga 2881 ggacctccta cccccttggc cccctcccct cccccagacc tccacctcct ccagaccccc 2941 tcccctcctg tgtcccttgt aaatagaacc agcccagccc gtctcctctt cccttccctg 3001 gcccccgggt gtaaatagat tgttataatt tttttcttaa agaaaacgtc gattcgcacc 3061 gtccaacctg ccccgcccct cctacagctg taactcccct cctgtcctct gcccccaagg 3121 tctactccct cctcacccca ccctggaggg ccaggggagt ggagagagct cctgatgtct 3181 tagtttccac agtaaggttt gcctgtgtac agacctccgt tcaataaatt attggcatga 3241 aaagaattcg gggggggggg ggggggggac agctcagggc tgcgatttcg cgccaaactt 3301 gacggcaatc ctagcgtgaa ggctggtagg attttatccc cgctgccatc atggttcgac 3361 cattgaactg catcgtcgcc gtgtcccaaa atatggggat tggcaagaac ggagacctac 3421 cctggcctcc gctcaggaac gagttcaagt acttccaaag aatgaccaca acctcttcag 3481 tggaaggtaa acagaatctg gtgattatgg gtaggaaaac ctggttctcc attcctgaga 3541 agaatcgacc tttaaaggac agaattaata tagttctcag tagagaactc aaagaaccac 3601 cacgaggagc tcattttctt gccaaaagtt tggatgatgc cttaagactt attgaacaac 3661 cggaattggc aagtaaagta gacatggttt ggatagtcgg aggcagttct gtttaccagg 3721 aagccatgaa tcaaccaggc cacctcagac tctttgtgac aaggatcatg caggaatttg 3781 aaagtgacac gtttttccca gaaattgatt tggggaaata taaacttctc ccagaatacc 3841 caggcgtcct ctctgaggtc caggaggaaa aaggcatcaa gtataagttt gaagtctacg 3901 agaagaaaga ctaacaggaa gatgctttca agttctctgc tcccctccta aagctatgca 3961 ttttttataa gaccatggga cttttgctgg ctttagatca taatcagcca taccacattt 4021 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa 4081 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc 4141 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 4201 tccaaactca tcaatgtatc ttatcatgtc tggatccccg gccaacggtc tggtgacccg 4261 gctgcgagag ctcggtgtac ctgagacgcg agtaagccct tgagtcaaag acgtagtcgt 4321 tgcaagtccg caccaggtac tgatatccca ccaaaaagtg cggcggcggc tggcggtaga 4381 ggggccagcg tagggtggcc ggggctccgg gggcgaggtc ttccaacata aggcgatgat 4441 catcgatgct agaccgtgca aaaggagagc ctgtaagcgg gcactcttcc gtggtctggt 4501 ggataaattc gcaagggtat catggcggac gaccggggtt cgaaccccgg atccggccgt 4561 ccgccgtgat ccatccggtt accgcccgcg tgtcgaaccc aggtgtgcga cgtcagacaa 4621 cgggggagcg ctccttttgg cttccttcca ggcgcggcgg ctgctgcgct agcttttttg 4681 gcgagctcga attaattctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 4741 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 4801 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 4861 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 4921 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 4981 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 5041 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 5101 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt 5161 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 5221 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 5281 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 5341 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 5401 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 5461 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 5521 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 5581 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 5641 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 5701 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 5761 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 5821 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 5881 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 5941 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 6001 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 6061 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 6121 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 6181 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 6241 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 6301 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 6361 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 6421 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 6481 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 6541 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 6601 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 6661 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct 6721 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa 6781 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga 6841 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact 6901 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca 6961 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt 7021 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg 7081 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga 7141 cggccagtgc c //