LOCUS Exported 7169 bp ds-DNA circular SYN 15-MAY-2020 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7169) AUTHORS HXN TITLE Direct Submission JOURNAL Exported Monday, May 18, 2020 from SnapGene Viewer 5.1.2 https://www.snapgene.com FEATURES Location/Qualifiers source 1..7169 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 1..130 /label=ITR misc_feature 143..823 /label=GFAP CDS 839..859 /codon_start=1 /product="nuclear localization signal of SV40 large T antigen" /label=NLS /translation="PKKKRKV" CDS 869..3766 /codon_start=1 /label=CasRx /translation="IEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDSI RSVNEGEAFSAEMADKNAGYKIGNAKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLEK RYFGESADGNDNICIQVIHNILDIEKILAEYITNAAYAVNNISGLDKDIIGFGKFSTVY TYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFSKEGRNYIIN YGNECYDILALLSGLRHWVVHNNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITNE LTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRKD MSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFVI NLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPAG RDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAFF KDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDEN GNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQK KQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGREN AEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLEE KGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKKA EEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFRNKAVHLEVARYVHA YINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCVP FGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNS" CDS 3776..3796 /codon_start=1 /product="nuclear localization signal of SV40 large T antigen" /label=NLS /translation="PKKKRKV" misc_feature 3806..3829 /label=FLAG polyA_signal 3842..3976 /label=SV40 polyA /note="SV40 polyadenylation signal" promoter 3983..4223 /label=U6 Promoter /note="RNA polymerase III promoter for human U6 snRNA" misc_feature 4233..4268 /label=DR misc_feature 4299..4334 /label=DR misc_feature 4365..4400 /label=DR repeat_region 4432..4572 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" rep_origin 4647..5102 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 5384..5488 /gene="bla" /label=AmpR promoter CDS 5489..6349 /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" rep_origin 6520..7108 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct gcggcctcta gaaacatatc ctggtgtgga gtaggggacg ctgctctgac 181 agaggctcgg gggcctgagc tggctctgtg agctggggag gaggcagaca gccaggcctt 241 gtctgcaagc agacctggca gcattgggct ggccgccccc cagggcctcc tcttcatgcc 301 cagtgaatga ctcaccttgg cacagacaca atgttcgggg tgggcacagt gcctgcttcc 361 cgccgcaccc cagcccccct caaatgcctt ccgagaagcc cattgagcag ggggcttgca 421 ttgcacccca gcctgacagc ctggcatctt gggataaaag cagcacagcc ccctaggggc 481 tgcccttgct gtgtggcgcc accggcggtg gagaacaagg ctctattcag cctgtgccca 541 ggaaagggga tcaggggatg cccaggcatg gacagtgggt ggcagggggg gagaggaggg 601 ctgtctgctt cccagaagtc caaggacaca aatgggtgag gggagagctc tccccatagc 661 tgggctgcgg cccaacccca ccccctcagg ctatgccagg gggtgttgcc aggggcaccc 721 gggcatcgcc agtctagccc actccttcat aaagccctcg catcccagga gcgagcagag 781 ccagagcagg ttggagagga gacgcatcac ctccgctgct cgcaccggtg ccaccatgcc 841 taaaaagaaa agaaaggtgg gttctggtat cgagaagaag aagagcttcg ccaagggcat 901 gggagtgaag agcaccctgg tgtccggctc taaggtgtac atgaccacat ttgctgaggg 961 aagcgacgcc aggctggaga agatcgtgga gggcgatagc atcagatccg tgaacgaggg 1021 agaggctttc agcgccgaga tggctgacaa gaacgctggc tacaagatcg gaaacgccaa 1081 gttttcccac ccaaagggct acgccgtggt ggctaacaac ccactgtaca ccggaccagt 1141 gcagcaggac atgctgggac tgaaggagac actggagaag aggtacttcg gcgagtccgc 1201 cgacggaaac gataacatct gcatccaggt catccacaac atcctggata tcgagaagat 1261 cctggctgag tacatcacaa acgccgctta cgccgtgaac aacatctccg gcctggacaa 1321 ggatatcatc ggcttcggaa agttttctac cgtgtacaca tacgacgagt tcaaggatcc 1381 agagcaccac cgggccgctt ttaacaacaa cgacaagctg atcaacgcca tcaaggctca 1441 gtacgacgag ttcgataact ttctggataa ccccaggctg ggctacttcg gacaggcttt 1501 cttttctaag gagggcagaa actacatcat caactacgga aacgagtgtt acgacatcct 1561 ggccctgctg agcggactga ggcactgggt ggtgcacaac aacgaggagg agtctcggat 1621 cagccgcacc tggctgtaca acctggacaa gaacctggat aacgagtaca tctccacact 1681 gaactacctg tacgacagga tcaccaacga gctgacaaac agcttctcca agaactctgc 1741 cgctaacgtg aactacatcg ctgagaccct gggcatcaac ccagctgagt tcgctgagca 1801 gtacttcaga ttttccatca tgaaggagca gaagaacctg ggcttcaaca tcacaaagct 1861 gagagaagtg atgctggaca gaaaggatat gtccgagatc aggaagaacc acaaggtgtt 1921 cgattctatc agaaccaagg tgtacacaat gatggacttt gtgatctaca ggtactacat 1981 cgaggaggat gccaaggtgg ccgctgccaa caagagcctg cccgacaacg agaagtctct 2041 gagcgagaag gatatcttcg tgatcaacct gagaggctcc tttaacgacg atcagaagga 2101 cgctctgtac tacgatgagg ccaacaggat ctggagaaag ctggagaaca tcatgcacaa 2161 catcaaggag ttccggggaa acaagacccg cgagtacaag aagaaggacg ctccaaggct 2221 gcctaggatc ctgcctgctg gaagggacgt gagcgccttc agcaagctga tgtacgccct 2281 gacaatgttt ctggacggaa aggagatcaa cgatctgctg accacactga tcaacaagtt 2341 cgacaacatc cagtcttttc tgaaagtgat gcctctgatc ggcgtgaacg ctaagttcgt 2401 ggaggagtac gccttcttta aggacagcgc caagatcgct gatgagctgc ggctgatcaa 2461 gtcctttgcc aggatgggag agccaatcgc tgacgctagg agagctatgt acatcgatgc 2521 catccggatc ctgggaacca acctgtctta cgacgagctg aaggctctgg ccgacacctt 2581 cagcctggat gagaacggca acaagctgaa gaagggcaag cacggaatgc gcaacttcat 2641 catcaacaac gtgatcagca acaagcggtt tcactacctg atcagatacg gcgacccagc 2701 tcacctgcac gagatcgcta agaacgaggc cgtggtgaag ttcgtgctgg gacggatcgc 2761 cgatatccag aagaagcagg gccagaacgg aaagaaccag atcgaccgct actacgagac 2821 ctgcatcggc aaggataagg gaaagtccgt gtctgagaag gtggacgctc tgaccaagat 2881 catcacaggc atgaactacg accagttcga taagaagaga tctgtgatcg aggacaccgg 2941 aagggagaac gccgagagag agaagtttaa gaagatcatc agcctgtacc tgacagtgat 3001 ctaccacatc ctgaagaaca tcgtgaacat caacgctaga tacgtgatcg gcttccactg 3061 cgtggagcgc gatgcccagc tgtacaagga gaagggatac gacatcaacc tgaagaagct 3121 ggaggagaag ggctttagct ccgtgaccaa gctgtgcgct ggaatcgacg agacagcccc 3181 cgacaagagg aaggatgtgg agaaggagat ggccgagaga gctaaggaga gcatcgactc 3241 cctggagtct gctaacccta agctgtacgc caactacatc aagtactccg atgagaagaa 3301 ggccgaggag ttcaccaggc agatcaacag agagaaggcc aagaccgctc tgaacgccta 3361 cctgaggaac acaaagtgga acgtgatcat ccgggaggac ctgctgcgca tcgataacaa 3421 gacctgtaca ctgttccgga acaaggctgt gcacctggag gtggctcgct acgtgcacgc 3481 ctacatcaac gacatcgccg aggtgaactc ctactttcag ctgtaccact acatcatgca 3541 gaggatcatc atgaacgaga gatacgagaa gtctagcggc aaggtgtctg agtacttcga 3601 cgccgtgaac gatgagaaga agtacaacga tagactgctg aagctgctgt gcgtgccttt 3661 cggatactgt atcccacggt ttaagaacct gagcatcgag gccctgttcg accgcaacga 3721 ggctgccaag tttgataagg agaagaagaa ggtgagcggc aactccggtt ctggtcccaa 3781 gaagaagcgc aaggtggcgg ccgctgatta caaagacgat gacgataagt aaagatccta 3841 aaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 3901 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 3961 ttatcatgtc tggatcgcgg ccgagggcct atttcccatg attccttcat atttgcatat 4021 acgatacaag gctgttagag agataattgg aattaatttg actgtaaaca caaagatatt 4081 agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt 4141 atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc 4201 tttatatatc ttgtggaaag gacgaaacac cgcaagtaaa cccctaccaa ctggtcgggg 4261 tttgaaactg tagatgggct gtccacgaag cactggcgca agtaaacccc taccaactgg 4321 tcggggtttg aaacgcttgg agaagtcgat gcgcagcgtg cagccaagta aacccctacc 4381 aactggtcgg ggtttgaaac tttttttgaa ttctgatgcg gtggcggccg caggaacccc 4441 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac 4501 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca 4561 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac 4621 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg 4681 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt 4741 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 4801 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 4861 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 4921 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc 4981 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa 5041 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat 5101 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca 5161 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag 5221 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa 5281 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat 5341 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg 5401 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 5461 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 5521 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 5581 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 5641 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 5701 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg 5761 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 5821 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 5881 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 5941 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 6001 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 6061 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 6121 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 6181 taaatctgga gccggtgagc gtggaagccg cggtatcatt gcagcactgg ggccagatgg 6241 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 6301 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 6361 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 6421 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 6481 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 6541 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 6601 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 6661 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 6721 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 6781 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 6841 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 6901 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 6961 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 7021 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 7081 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 7141 ggccttttgc tggccttttg ctcacatgt //