LOCUS Exported 7602 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pSico p53 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7602) AUTHORS Ventura A, Meissner A, Dillon CP, McManus M, Sharp PA, Van Parijs L, Jaenisch R, Jacks T TITLE Cre-lox-regulated conditional RNA interference from transgenes. JOURNAL Proc Natl Acad Sci U S A. 2004 Jul 13. 101(28):10380-5. PUBMED 15240889 REFERENCE 2 (bases 1 to 7602) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7602 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(47..66) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 238..617 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 619..817 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" LTR 835..1015 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1062..1187 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1684..1917 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2102..2146 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 2444..2561 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" primer_bind 2851..2871 /label=mU6-F /note="Mouse U6 promoter, forward primer" protein_bind 2889..2922 /label=loxP511 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GTATACAT). loxP511 sites are compatible with each other, but incompatible with wild-type loxP sites." enhancer 3022..3325 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3326..3529 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" regulatory 3568..3577 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3574..4290 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3619..3640) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3880..3899) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4227..4248 /label=EGFP-C /note="EGFP, forward primer" protein_bind 4301..4334 /label=loxP511 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GTATACAT). loxP511 sites are compatible with each other, but incompatible with wild-type loxP sites." CDS complement(4353..4364) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" CDS 4374..4385 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind 4401..4417 /label=KS primer /note="common sequencing primer, one of multiple similar variants" misc_feature 4492..5080 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4545..4565) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4963..4974) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(5083..5099) /label=KS primer /note="common sequencing primer, one of multiple similar variants" LTR 5608..5788 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(5850..6438) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5930..5949) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6609..7469) /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 7232..7251 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7470..7574) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg 61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac 241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt 661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg 781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct 841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc 1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc 1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa 1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg 1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata 1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc 1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga 1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc 1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca 1501 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga 1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt 1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg 1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc 1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa 1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa 1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg 1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg 1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga 2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga 2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa 2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg 2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc 2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga 2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact 2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg 2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa 2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca 2581 gcagagatcc agtttggtta gtaccgggcc cgctctagag atccgacgcg ccatctctag 2641 gcccgcgccg gccccctcgc acagacttgt gggagaagct cggctactcc cctgccccgg 2701 ttaatttgca tataatattt cctagtaact atagaggctt aatgtgcgat aaaagacaga 2761 taatctgttc tttttaatac tagctacatt ttacatgata ggcttggatt tctataagag 2821 atacaaatac taaattatta ttttaaaaaa cagcacaaaa ggaaactcac cctaactgta 2881 aagtaattat aacttcgtat agtataaatt atacgaagtt ataagccttg gttttttgaa 2941 ttccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca 3001 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc 3061 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat 3121 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt 3181 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc 3241 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta 3301 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg 3361 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt 3421 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac 3481 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa 3541 ccgtcagatc cgctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc 3601 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc 3661 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc 3721 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg 3781 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg 3841 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc 3901 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc 3961 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac 4021 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc 4081 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc 4141 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc 4201 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg 4261 atcactctcg gcatggacga gctgtacaag tagcggccgc ataacttcgt atagtataaa 4321 ttatacgaag ttataagcct tgtttgtact ctcctcccct caatttcaag agaattgagg 4381 ggaggagagt acttttttct cgaggtcgac ggtatcgata agctcgcttc acgagatcat 4441 gtttaagggt tccggttcca ctaggtacaa ttcgatatca agcttatcga taatcaacct 4501 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 4561 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 4621 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 4681 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 4741 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 4801 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 4861 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 4921 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 4981 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 5041 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga 5101 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga 5161 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt 5221 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa 5281 ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat 5341 ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc cagggatcag 5401 atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga 5461 agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga 5521 tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat 5581 ggcccgagag ctgcatccgg actgtatggg tctctctggt tagaccagat ctgagcctgg 5641 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg 5701 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc 5761 ttttagtcag tgtggaaaat ctctagcagc atgtgagcaa aaggccagca aaaggccagg 5821 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 5881 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 5941 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 6001 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 6061 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 6121 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 6181 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 6241 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 6301 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 6361 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 6421 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 6481 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 6541 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 6601 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 6661 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 6721 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 6781 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 6841 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 6901 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 6961 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 7021 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 7081 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 7141 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 7201 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 7261 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 7321 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 7381 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 7441 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 7501 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 7561 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ac //