LOCUS Exported 5466 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses the C-terminal split-intein fragment of S. aureus BE3.9max from the CMV promoter. ACCESSION . VERSION . KEYWORDS CMV_Npu-saBE3.9max C-terminal SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5466) AUTHORS Levy JM, Yeh WH, Pendse N, Davis JR, Hennessey E, Butcher R, Koblan LW, Comander J, Liu Q, Liu DR TITLE Cytosine and adenine base editing of the brain, liver, retina, heart and skeletal muscle of mice via adeno-associated viruses. JOURNAL Nat Biomed Eng. 2020 Jan;4(1):97-110. doi: 10.1038/s41551-019-0501-5. Epub 2020 Jan 14. PUBMED 31937940 REFERENCE 2 (bases 1 to 5466) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5466 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 132..335 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 286..306 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 377..396 /label=T7 /note="T7 promoter, forward primer" promoter 377..395 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 445..465 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2161..2409 /codon_start=1 /product="uracil-DNA glycosylase inhibitor from a Bacillus subtilis bacteriophage (Mol et al., 1995)" /label=UGI /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML" CDS 2452..2472 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2481..2498 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(2521..2538) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2527..2751 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(2822..2838) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2822..2838) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2835..2857) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2846..2862 /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)." promoter complement(2870..2900) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2915..2936 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3053..3070) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3224..3812) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3304..3323) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3983..4843) /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 4606..4625 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4844..4948) /gene="bla" /label=AmpR promoter primer_bind complement(5027..5046) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 5218..131 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" ORIGIN 1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg 61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg 121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact 181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa 241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta 301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct 361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat gaaacggaca 421 gccgacggaa gcgagttcga gtcaccaaag aagaagcgga aagtcatcaa gattgctaca 481 cggaaatacc tgggaaagca gaacgtgtac gacatcggcg tggagcggga tcacaacttc 541 gccctgaaga atggctttat cgccagcaat tgtctgtatt ctctggaggc catccctctg 601 gaggacctgc tgaacaatcc attcaactac gaggtggatc acatcatccc ccggagcgtg 661 agcttcgaca attcttttaa caataaggtg ctggtgaagc aggaggagaa cagcaagaag 721 ggcaatagga cccctttcca gtacctgtct agctccgatt ctaagatcag ctacgagaca 781 ttcaagaagc acatcctgaa tctggccaag ggcaagggcc gcatcagcaa gaccaagaag 841 gagtacctgc tggaggagcg ggacatcaac agattctccg tgcagaagga cttcatcaac 901 cggaatctgg tggacaccag atacgccaca cgcggcctga tgaatctgct gcggtcttat 961 ttcagagtga acaatctgga tgtgaaggtg aagagcatca acggcggctt cacctccttt 1021 ctgcggagaa agtggaagtt taagaaggag cgcaacaagg gctataagca ccacgccgag 1081 gatgccctga tcatcgccaa tgccgacttc atctttaagg agtggaagaa gctggacaag 1141 gccaagaaag tgatggagaa ccagatgttc gaggagaagc aggccgagag catgcccgag 1201 atcgagacag agcaggagta caaggagatt ttcatcacac ctcaccagat caagcacatc 1261 aaggacttca aggactacaa gtattctcac agggtggata agaagcccaa ccgcgagctg 1321 atcaatgaca ccctgtatag cacacggaag gacgataagg gcaataccct gatcgtgaac 1381 aatctgaacg gcctgtacga caaggataat gacaagctga agaagctgat caacaagtct 1441 cccgagaagc tgctgatgta ccaccacgat cctcagacat atcagaagct gaagctgatc 1501 atggagcagt acggcgacga gaagaaccca ctgtataagt actatgagga gacaggcaac 1561 tacctgacaa agtatagcaa gaaggataat ggccccgtga tcaagaagat caagtactat 1621 ggcaacaagc tgaatgccca cctggacatc accgacgatt accctaactc tcgcaataag 1681 gtggtgaagc tgagcctgaa gccataccgg ttcgacgtgt acctggacaa cggcgtgtat 1741 aagtttgtga cagtgaagaa tctggatgtg atcaagaagg agaactacta tgaggtgaac 1801 agcaagtgct acgaggaggc caagaagctg aagaagatca gcaaccaggc cgagttcatc 1861 gcctcttttt acaacaatga cctgatcaag atcaatggcg agctgtatag agtgatcggc 1921 gtgaacaatg atctgctgaa cagaatcgaa gtgaatatga tcgacatcac ctacagggag 1981 tatctggaga acatgaatga taagaggccc cctcgcatca tcaagaccat cgcctctaag 2041 acacagagca tcaagaagta cagcacagac atcctgggga acctgtatga agtcaagagc 2101 aagaaacatc ctcagattat caagaaaggc agcggcggga gcggcgggag cggggggagc 2161 actaatctga gcgacatcat tgagaaggag actgggaaac agctggtcat tcaggagtcc 2221 atcctgatgc tgcctgagga ggtggaggaa gtgatcggca acaagccaga gtctgacatc 2281 ctggtgcaca ccgcctacga cgagtccaca gatgagaatg tgatgctgct gacctctgac 2341 gcccccgagt ataagccttg ggccctggtc atccaggatt ctaacggcga gaataagatc 2401 aagatgctga gcggaggatc caaaagaacc gccgacggca gcgaattcga gcccaagaag 2461 aagaggaaag tctaaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag 2521 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct 2581 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc 2641 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg 2701 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg 2761 cggaaagaac cagctggggc tcgataccgt cgacctctag ctagagcttg gcgtaatcat 2821 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 2881 ccggaagcat aaagtgtaaa gcctagggtg cctaatgagt gagctaactc acattaattg 2941 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 3001 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 3061 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 3121 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 3181 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 3241 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 3301 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 3361 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 3421 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 3481 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 3541 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 3601 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 3661 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 3721 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 3781 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 3841 ctgacactca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 3901 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 3961 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 4021 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 4081 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 4141 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 4201 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 4261 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 4321 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 4381 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 4441 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 4501 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 4561 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 4621 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 4681 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 4741 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 4801 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 4861 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 4921 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg 4981 acggatcggg agatcgatct cccgatcccc tagggtcgac tctcagtaca atctgctctg 5041 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 5101 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 5161 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac 5221 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 5281 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 5341 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 5401 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 5461 tgtatc //