LOCUS Exported 8462 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBMN ER50-YFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8462) AUTHORS Miyazaki Y, Imoto H, Chen LC, Wandless TJ TITLE Destabilizing domains derived from the human estrogen receptor. JOURNAL J Am Chem Soc. 2012 Mar 7;134(9):3942-5. Epub 2012 Feb 22. PUBMED 22332638 REFERENCE 2 (bases 1 to 8462) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8462 /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 2637..3350 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(2679..2700) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2940..2959) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3287..3308 /label=EGFP-C /note="EGFP, forward primer" CDS 3357..3383 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" misc_feature 3432..4017 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(3599..3616) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 3826..3845 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 4018..4701 /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 4714..5400 /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 5449..6042 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(6173..6190) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6344..6932) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6424..6443) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7106..7966) /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 7729..7748 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" primer_bind 8034..8052 /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 ccatgagcct tgccctgtca cttacagccg 1921 accagatggt ttccgcgctt ctcgacgctg aacctccaat tctctattcc gaatacgacc 1981 caaccaggcc gttctccgag gcatctatga tgggtctgct gacaaatctg gcagacaggg 2041 aactggtgca catgatcaat tgggcgaagc gcgtacccgg attcgtcgat cttgcactcc 2101 atgatcaggt gcacttgctg gagtgcgctt ggatggagat cctcatgatc gggctggtgt 2161 ggcggagtat ggaacacccc ggcaagttgc tgtttgcgcc taacctcctg ttggacagga 2221 accaggggaa atgtgtggag ggcggtgtgg aaatctttga catgctcctc gctacctcaa 2281 gccggtttag gatgatgaat ctgcagggcg aagagttcgt gtgtctcaaa tctatcatac 2341 tgttgaacag cggagtctac accttcctct ccagtactct gaaatctctg gaggagaaag 2401 atcatatcca tcgcgtgctg gacaagataa ccgacacgtt gattcacttg atggccaaag 2461 ctgggctcac tctgcaacaa caacatcagc gactggcaca gctgttgctg attttgagcc 2521 acattcggca catgtccagc aagagaatgg agcacctcta tagtatgaag tgcaagaacg 2581 tcgtacccct gtcagatctg cttcttgaaa tgcttgatgc ccaccggctg gaattcgtga 2641 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg 2701 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc 2761 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga 2821 ccaccttcgg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg 2881 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg 2941 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc 3001 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg 3061 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca 3121 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact 3181 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga 3241 gctaccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg 3301 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag gtcgactatc 3361 cgtacgacgt accagactac gcactcgact aactcgaggg ccggcgcgcc gcggccgcta 3421 cgtaaattcc gcccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa 3481 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat 3541 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct 3601 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct 3661 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc 3721 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa 3781 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc 3841 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg 3901 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaacgt ctaggccccc 3961 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg 4021 gtgagcggcc tgctgaagga gagtatgcgc atcaagatgt acatggaggg caccgtgaac 4081 ggccactact tcaagtgcga gggcgagggc gacggcaacc ccttcgccgg cacccagagc 4141 atgagaatcc acgtgaccga gggcgccccc ctgcccttcg ccttcgacat cctggccccc 4201 tgctgcgagt acggcagcag gaccttcgtg caccacaccg ccgagatccc cgacttcttc 4261 aagcagagct tccccgaggg cttcacctgg gagagaacca ccacctacga ggacggcggc 4321 atcctgaccg cccaccagga caccagcctg gagggcaact gcctgatcta caaggtgaag 4381 gtgcacggca ccaacttccc cgccgacggc cccgtgatga agaacaagag cggcggctgg 4441 gagcccagca ccgaggtggt gtaccccgag aacggcgtgc tgtgcggccg gaacgtgatg 4501 gccctgaagg tgggcgaccg gcacctgatc tgccaccact acaccagcta ccggagcaag 4561 aaggccgtgc gcgccctgac catgcccggc ttccacttca ccgacatccg gctccagatg 4621 ctgcggaaga agaaggacga gtacttcgag ctgtacgagg ccagcgtggc ccggtacagc 4681 gacctgcccg agaaggccaa cagatctccc gggatggtga gcggcctgct gaaggagagt 4741 atgcgcatca agatgtacat ggagggcacc gtgaacggcc actacttcaa gtgcgagggc 4801 gagggcgacg gcaacccctt cgccggcacc cagagcatga gaatccacgt gaccgagggc 4861 gcccccctgc ccttcgcctt cgacatcctg gccccctgct gcgagtacgg cagcaggacc 4921 ttcgtgcacc acaccgccga gatccccgac ttcttcaagc agagcttccc cgagggcttc 4981 acctgggaga gaaccaccac ctacgaggac ggcggcatcc tgaccgccca ccaggacacc 5041 agcctggagg gcaactgcct gatctacaag gtgaaggtgc acggcaccaa cttccccgcc 5101 gacggccccg tgatgaagaa caagagcggc ggctgggagc ccagcaccga ggtggtgtac 5161 cccgagaacg gcgtgctgtg cggccggaac gtgatggccc tgaaggtggg cgaccggcac 5221 ctgatctgcc accactacac cagctaccgg agcaagaagg ccgtgcgcgc cctgaccatg 5281 cccggcttcc acttcaccga catccggctc cagatgctgc ggaagaagaa ggacgagtac 5341 ttcgagctgt acgaggccag cgtggcccgg tacagcgacc tgcccgagaa ggccaactga 5401 gtcgacgata aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc 5461 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaaatac 5521 ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag ctgaatatgg 5581 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg 5641 aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg 5701 gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag agaaccatca 5761 gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa 5821 tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc 5881 acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta 5941 tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc 6001 tcctctgagt gattgactac ccgtcagcgg gggtctttca ttcacatgca gcatgtatca 6061 aaattaattt ggtttttttt cttaagtatt tacattaaat ggccatagtt gcattaatga 6121 atcggccaac gcgcggggag aggcggtttg cgtattggcg ctcttccgct tcctcgctca 6181 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 6241 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 6301 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 6361 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 6421 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 6481 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 6541 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 6601 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 6661 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 6721 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 6781 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 6841 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 6901 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 6961 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 7021 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt gcgcaaatca atctaaagta 7081 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 7141 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 7201 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 7261 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 7321 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 7381 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 7441 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 7501 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 7561 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 7621 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 7681 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc 7741 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 7801 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 7861 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 7921 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 7981 aatattattg aagcatttat cagggttatt gtctcatgac attaacctat aaaaataggc 8041 gtatcacgag gccctttcgt cttcaagaac agctttgctc ttaggagttt cctaatacat 8101 cccaaactca aatatataaa gcatttgact tgttctatgc cctagggggc ggggggaagc 8161 taagccagct ttttttaaca tttaaaatgt taattccatt ttaaatgcac agatgttttt 8221 atttcataag ggtttcaatg tgcatgaatg ctgcaatatt cctgttacca aagctagtat 8281 aaataaaaat agataaacgt ggaaattact tagagtttct gtcattaacg tttccttcct 8341 cagttgacaa cataaatgcg ctgctgagaa gccagtttgc atctgtcagg atcaatttcc 8401 cattatgcca gtcatattaa ttactagtca attagttgat ttttattttt gacatataca 8461 tg //