LOCUS Exported 8176 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBMN YFP-DHFR(DD) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8176) AUTHORS Iwamoto M, Bjoerklund T, Lundberg C, Kirik D, Wandless TJ TITLE A general chemical method to regulate protein stability in the mammalian central nervous system. JOURNAL Chem Biol. 2010 Sep 24. 17(9):981-8. PUBMED 20851347 REFERENCE 2 (bases 1 to 8176) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8176 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 1..594 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 657..1014 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 1079..1495 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1425..1447 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1463..1479 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" misc_feature 1505..1878 /label=pol region /note="Moloney murine leukemia virus (MMLV) pol region containing the splice acceptor site" primer_bind 1816..1832 /label=pBMN 5' /note="MMLV sequence, for inserts in pBMN retroviral vector" CDS 1896..1922 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1923..2639 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1968..1989) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2229..2248) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2576..2597 /label=EGFP-C /note="EGFP, forward primer" CDS 2646..3122 /codon_start=1 /gene="folA" /product="E. coli dihydrofolate reductase" /label=ecDHFR /translation="ISLIAALAVDHVIGMETVMPWNLPADLAWFKRNTLNKPVIMGRHT WESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLP KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR" misc_feature 3146..3731 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(3313..3330) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 3540..3559 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 3732..4415 /codon_start=1 /product="far-red fluorescent variant of Heteractis crispa chromoprotein" /label=HcRed1 /note="mammalian codon-optimized" /translation="MVSGLLKESMRIKMYMEGTVNGHYFKCEGEGDGNPFAGTQSMRIH VTEGAPLPFAFDILAPCCEYGSRTFVHHTAEIPDFFKQSFPEGFTWERTTTYEDGGILT AHQDTSLEGNCLIYKVKVHGTNFPADGPVMKNKSGGWEPSTEVVYPENGVLCGRNVMAL KVGDRHLICHHYTSYRSKKAVRALTMPGFHFTDIRLQMLRKKKDEYFELYEASVARYSD LPEKAN" CDS 4428..5114 /codon_start=1 /product="far-red fluorescent variant of Heteractis crispa chromoprotein" /label=HcRed1 /note="mammalian codon-optimized" /translation="MVSGLLKESMRIKMYMEGTVNGHYFKCEGEGDGNPFAGTQSMRIH VTEGAPLPFAFDILAPCCEYGSRTFVHHTAEIPDFFKQSFPEGFTWERTTTYEDGGILT AHQDTSLEGNCLIYKVKVHGTNFPADGPVMKNKSGGWEPSTEVVYPENGVLCGRNVMAL KVGDRHLICHHYTSYRSKKAVRALTMPGFHFTDIRLQMLRKKKDEYFELYEASVARYSD LPEKAN" LTR 5163..5756 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(5887..5904) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6058..6646) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6138..6157) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6820..7680) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 7443..7462 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" primer_bind 7748..7766 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc 61 atggaaaaat acataactga gaatagaaaa gttcagatca aggtcaggaa cagatggaac 121 agctgaatat gggccaaagc ggatatctgt ggtaagcagt tcctgccccg gctcagggcc 181 aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg 241 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct 301 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt 361 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca 421 ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg 481 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc 541 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg 601 ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc 661 tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgactga ttttatgcgc 721 ctgcgtcggt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac 781 gagttcggaa cacccggccg caaccctggg agacgtccca gggacttcgg gggccgtttt 841 tgtggcccga cctgagtcca aaaatcccga tcgttttgga ctctttggtg cacccccctt 901 agaggaggga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc 961 tgaatttttg ctttcggttt gggaccgaag ccgcgccgcg cgtcttgtct gctgcagcat 1021 cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaata tgggcccggg 1081 ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat 1141 cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga 1201 atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac 1261 ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta 1321 catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca 1381 ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg 1441 ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag gcgcccccat 1501 atggccatat gagatcttat atggggcacc cccgcccctt gtaaacttcc ctgaccctga 1561 catgacaaga gttactaaca gcccctctct ccaagctcac ttacaggctc tctacttagt 1621 ccagcacgaa gtctggagac ctctggcggc acgtaccaag aacaactgga ccgaccggtg 1681 gtacctcacc cttaccgagt cggcgacaca gtgtgggtcc gccgacacca gactaagaac 1741 ctagaacctc gctggaaagg accttacaca gtcctgctga ccacccccac cgccctcaaa 1801 gtagacggca tcgcagcttg gatacacgcc gcccacgtga aggctgccga ccccgggggt 1861 ggaccatcct ctagactgcc ggatccgcca ccatgtatcc gtacgacgta ccggattatg 1921 cgatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg 1981 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct 2041 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca 2101 ccctcgtgac caccttcggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga 2161 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct 2221 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc 2281 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc 2341 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga 2401 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg 2461 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc 2521 actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg 2581 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagc 2641 tcgagatcag tctgattgcg gcgttagcgg tagatcacgt tatcggcatg gaaaccgtca 2701 tgccgtggaa cctgcctgcc gatctcgcct ggtttaaacg caacacctta aataaacccg 2761 tgattatggg ccgccatacc tgggaatcaa tcggtcgtcc gttgccagga cgcaaaaata 2821 ttatcctcag cagtcaaccg agtacggacg atcgcgtaac gtgggtgaag tcggtggatg 2881 aagccatcgc ggcgtgtggt gacgtaccag aaatcatggt tattggcggc ggtcgcgttt 2941 atgaacagtt cttgccaaaa gcgcaaaaac tgtatctgac gcatatcgac gcagaagtgg 3001 aaggcgacac ccatttcccg gattacgagc cggatgactg ggaatcggta ttcagcgaat 3061 tccacgatgc tgatgcgcag aactctcaca gctattgctt tgagattctg gagcggcgat 3121 gaccgcggcc gctacgtaaa ttccgcccct ctccctcccc cccccctaac gttactggcc 3181 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc 3241 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta 3301 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag 3361 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga 3421 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg 3481 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat 3541 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta 3601 tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa 3661 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata 3721 tggccacaac catggtgagc ggcctgctga aggagagtat gcgcatcaag atgtacatgg 3781 agggcaccgt gaacggccac tacttcaagt gcgagggcga gggcgacggc aaccccttcg 3841 ccggcaccca gagcatgaga atccacgtga ccgagggcgc ccccctgccc ttcgccttcg 3901 acatcctggc cccctgctgc gagtacggca gcaggacctt cgtgcaccac accgccgaga 3961 tccccgactt cttcaagcag agcttccccg agggcttcac ctgggagaga accaccacct 4021 acgaggacgg cggcatcctg accgcccacc aggacaccag cctggagggc aactgcctga 4081 tctacaaggt gaaggtgcac ggcaccaact tccccgccga cggccccgtg atgaagaaca 4141 agagcggcgg ctgggagccc agcaccgagg tggtgtaccc cgagaacggc gtgctgtgcg 4201 gccggaacgt gatggccctg aaggtgggcg accggcacct gatctgccac cactacacca 4261 gctaccggag caagaaggcc gtgcgcgccc tgaccatgcc cggcttccac ttcaccgaca 4321 tccggctcca gatgctgcgg aagaagaagg acgagtactt cgagctgtac gaggccagcg 4381 tggcccggta cagcgacctg cccgagaagg ccaacagatc tcccgggatg gtgagcggcc 4441 tgctgaagga gagtatgcgc atcaagatgt acatggaggg caccgtgaac ggccactact 4501 tcaagtgcga gggcgagggc gacggcaacc ccttcgccgg cacccagagc atgagaatcc 4561 acgtgaccga gggcgccccc ctgcccttcg ccttcgacat cctggccccc tgctgcgagt 4621 acggcagcag gaccttcgtg caccacaccg ccgagatccc cgacttcttc aagcagagct 4681 tccccgaggg cttcacctgg gagagaacca ccacctacga ggacggcggc atcctgaccg 4741 cccaccagga caccagcctg gagggcaact gcctgatcta caaggtgaag gtgcacggca 4801 ccaacttccc cgccgacggc cccgtgatga agaacaagag cggcggctgg gagcccagca 4861 ccgaggtggt gtaccccgag aacggcgtgc tgtgcggccg gaacgtgatg gccctgaagg 4921 tgggcgaccg gcacctgatc tgccaccact acaccagcta ccggagcaag aaggccgtgc 4981 gcgccctgac catgcccggc ttccacttca ccgacatccg gctccagatg ctgcggaaga 5041 agaaggacga gtacttcgag ctgtacgagg ccagcgtggc ccggtacagc gacctgcccg 5101 agaaggccaa ctgagtcgac gataaaataa aagattttat ttagtctcca gaaaaagggg 5161 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag 5221 gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg aacagatgga 5281 acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg 5341 ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc 5401 tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc tcagcagttt 5461 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta 5521 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct 5581 caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc 5641 cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt 5701 tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcattcaca 5761 tgcagcatgt atcaaaatta atttggtttt ttttcttaag tatttacatt aaatggccat 5821 agttgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt ggcgctcttc 5881 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc 5941 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat 6001 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 6061 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 6121 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 6181 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt 6241 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 6301 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta 6361 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa 6421 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa 6481 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt 6541 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt 6601 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 6661 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 6721 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gtttgcgcaa 6781 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 6841 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 6901 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 6961 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 7021 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 7081 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 7141 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 7201 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 7261 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 7321 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 7381 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg 7441 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 7501 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 7561 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 7621 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 7681 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgacattaac 7741 ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaacagcttt gctcttagga 7801 gtttcctaat acatcccaaa ctcaaatata taaagcattt gacttgttct atgccctagg 7861 gggcgggggg aagctaagcc agcttttttt aacatttaaa atgttaattc cattttaaat 7921 gcacagatgt ttttatttca taagggtttc aatgtgcatg aatgctgcaa tattcctgtt 7981 accaaagcta gtataaataa aaatagataa acgtggaaat tacttagagt ttctgtcatt 8041 aacgtttcct tcctcagttg acaacataaa tgcgctgctg agaagccagt ttgcatctgt 8101 caggatcaat ttcccattat gccagtcata ttaattacta gtcaattagt tgatttttat 8161 ttttgacata tacatg //