LOCUS       Exported                7598 bp ds-DNA     circular SYN 29-JUN-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7598)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jan 2, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7598
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          438..934
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     source          4495..4570
                     /organism="Cloning vector M13KO7"
                     /mol_type="other DNA"
     source          4495..4570
                     /organism="Cloning vector M13KO7"
                     /mol_type="other DNA"
     source          4495..4570
                     /organism="Cloning vector M13KO7"
                     /mol_type="other DNA"
     source          4495..4570
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    156..368
                     /label=EFS
     promoter        156..367
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             384..437
                     /codon_start=1
                     /label=BPSV40 NLS
                     /translation="KRTADGSEFESPKKKRKV"
     CDS             438..935
                     /codon_start=1
                     /standard_name="max10"
                     /label=envTadA
                     /label=max10
                     /translation="SEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWN
                     RAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFG
                     VRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQS
                     SIN"
     misc_feature    936..1031
                     /label=linker
     CDS             1032..4190
                     /codon_start=1
                     /label=SlugCas9-NNG(D10A)
                     /translation="NQKFILGLAIGITSVGYGLIDYETKNIIDAGVRLFPEANVENNEG
                     RRSKRGSRRLKRRRIHRLERVKKLLEDYNLLDQSQIPQSTNPYAIRVKGLSEALSKDEL
                     VIALLHIAKRRGIHKIDVIDSNDDVGNELSTKEQLNKNSKLLKDKFVCQIQLERMNEGQ
                     VRGEKNRFKTADIIKEIIQLLNVQKNFHQLDENFINKYIELVEMRREYFEGPGKGSPYG
                     WEGDPKAWYETLMGHCTYFPDELRSVKYAYSADLFNALNDLNNLVIQRDGLSKLEYHEK
                     YHIIENVFKQKKKPTLKQIANEINVNPEDIKGYRITKSGKPQFTEFKLYHDLKSVLFDQ
                     SILENEDVLDQIAEILTIYQDKDSIKSKLTELDILLNEEDKENIAQLTGYTGTHRLSLK
                     CIRLVLEEQWYSSRNQMEIFTHLNIKPKKINLTAANKIPKAMIDEFILSPVVKRTFGQA
                     INLINKIIEKYGVPEDIIIELARENNSKDKQKFINEMQKKNENTRKRINEIIGKYGNQN
                     AKRLVEKIRLHDEQEGKCLYSLESIPLEDLLNNPNHYEVDHIIPRSVSFDNSYHNKVLV
                     KQSENSKKSNLTPYQYFNSGKSKLSYNQFKQHILNLSKSQDRISKKKKEYLLEERDINK
                     FEVQKEFINRNLVDTRYATRELTNYLKAYFSANNMNVKVKTINGSFTDYLRKVWKFKKE
                     RNHGYKHHAEDALIIANADFLFKENKKLKAVNSVLEKPEIESKQLDIQVDSEDNYSEMF
                     IIPKQVQDIKDFRNFKYSHRVDKKPNRRLINDTLYSTRKKDNSTYIVQTIKDIYAKDNT
                     TLKKQFDKSPEKFLMYQHDPRTFEKLEVIMKQYANEKNPLAKYHEETGEYLTKYSKKNN
                     GPIVKSLKYIGNKLGRHLDVTHQFKSSTKKLVKRSIKPYRFDVYLTDKGYKFITISYLD
                     VLKKDNYYYIPEQKYDKLKLGKAIDKNAKFIASFYKNDLIKLDGEIYKIIGVSSDTRNM
                     IELDLPDIRYKEYCELNNIKGKPRIIKTIGKKVNSIEKLTTDVLGNVFTNTQYTKPQLL
                     FKRGN"
     misc_feature    3372..3374
                     /label=Q782R
     misc_feature    3690..3691
                     /label=S888R
     misc_feature    3744..3746
                     /label=L906R
     misc_feature    3978..3980
                     /label=N984S
     misc_feature    4063..4064
                     /label=E1012K
     misc_feature    4074..4076
                     /label=K1016I
     CDS             4233..4253
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4263..4487
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    4495..4570
                     /label=Feature 48
     misc_feature    complement(4495..4570)
                     /label=Sa-trac
     promoter        complement(4590..4838)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    4839..4844
                     /label=Feature 11
     repeat_region   4861..5001
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      5076..5531
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5813..5917
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5918..6778
                     /label=amp
     CDS             5918..6778
                     /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      6949..7537
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgggca gagcgcacat cgcccacagt
      181 ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg
      241 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
      301 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
      361 aacacaggac cggtgccacc atgaaacgga cagccgacgg aagcgagttc gagtcaccaa
      421 agaagaagcg gaaagtctct gaggtggagt tttcccacga gtactggatg agacatgccc
      481 tgaccctggc caagagggca cgggatgaga gggaggtgcc tgtgggagcc gtgctggtgc
      541 tgaacaatag agtgatcggc gagggctgga acagagccat cggcctgcac gacccaacag
      601 cccatgccga aattatggcc ctgagacagg gcggcctggt catgcagaac tacagactga
      661 ttgacgccac cctgtacgtg acattcgagc cttgcgtgat gtgcgccggc gccatgatcc
      721 actctaggat cggccgcgtg gtgtttggcg tgaggaactc aaaaagaggc gccgcaggct
      781 ccctgatgaa cgtgctgaac taccccggca tgaatcaccg cgtcgaaatt accgagggaa
      841 tcctggcaga tgaatgtgcc gccctgctgt gcgatttcta tcggatgcct agacaggtgt
      901 tcaatgctca gaagaaggcc cagagctcca tcaactccgg aggatctagc ggaggctcct
      961 ctggctctga gacacctggc acaagcgaga gcgcaacacc tgaaagcagc gggggcagca
     1021 gcggggggtc aaaccaaaaa ttcatactgg gactggccat cggaatcacc agcgtgggct
     1081 acggcctgat cgactacgag acaaagaata tcatcgatgc cggcgttaga ctgttccccg
     1141 aggccaacgt ggaaaacaac gagggaagaa ggtccaaacg tggaagcaga agactgaagc
     1201 gacgccgcat tcacagactt gaacgggtga agaagctgct cgaggattat aatctgctgg
     1261 atcagtccca gattcctcag tctacaaacc cctacgccat ccgcgtgaag ggcctgtctg
     1321 aagccctgag caaggacgaa ctcgtgattg ccctgctcca tatcgccaag agaagaggca
     1381 tccacaagat cgacgtgatc gacagcaacg acgacgtggg gaacgagctc agcaccaagg
     1441 aacagctgaa taagaacagc aagctgctga aagacaaatt tgtgtgccag atccagctgg
     1501 aaagaatgaa tgagggccag gtgcggggag agaaaaaccg gttcaagacc gctgatatca
     1561 tcaaggaaat catccagctg ctgaatgtgc agaagaactt ccaccagctg gacgagaact
     1621 tcatcaacaa gtacatcgaa ctggttgaga tgagacggga atacttcgag ggccccggca
     1681 agggcagtcc atatggctgg gaaggcgacc ctaaggcttg gtacgagaca ctgatgggcc
     1741 actgcaccta cttcccagat gagctgagaa gcgtgaaata cgcctacagt gccgacctgt
     1801 tcaacgctct gaacgacctg aacaacctgg tcatccaaag agatggactg tctaagctcg
     1861 agtatcatga gaagtatcac atcatcgaga acgtgttcaa gcagaagaag aaacctacac
     1921 tgaagcagat cgccaatgag atcaatgtca accctgaaga tatcaagggc tacagaatca
     1981 caaagtctgg caagccccag tttaccgagt ttaagctcta ccacgacctg aaaagcgtgc
     2041 tgtttgacca gagcatcctg gagaacgaag acgtgctgga ccagatcgct gagatcctga
     2101 ccatctacca ggacaaggat agcatcaaat ctaagctgac ggaactggac atcctgctga
     2161 acgaggaaga taaggaaaac atcgcccagc tgactggcta caccgggacc caccggctca
     2221 gcctgaaatg catccggctg gtcctggaag agcagtggta ttctagccgg aaccagatgg
     2281 aaatcttcac ccacctgaac attaagccta agaagatcaa cctgacagcc gccaacaaga
     2341 tcccgaaggc tatgatcgac gagttcatcc tgagccctgt ggtgaagagg accttcggcc
     2401 aggccattaa ccttattaac aagatcatag aaaagtacgg cgtgcctgaa gatatcatca
     2461 tcgagctggc cagagaaaat aatagcaagg acaagcagaa gttcatcaat gagatgcaga
     2521 aaaagaacga gaacaccaga aagagaatta acgaaatcat cggcaagtat ggcaaccaga
     2581 acgccaagag actggtcgag aagattagac tgcacgacga gcaggagggc aagtgcctgt
     2641 actcactgga aagcatccct ctggaggacc tgctgaacaa ccccaaccac tacgaggtgg
     2701 accacatcat tccaagatct gtgtccttcg acaactctta ccacaacaaa gtgctcgtga
     2761 agcagagcga gaactccaaa aaatccaacc tgacccctta ccagtacttt aacagcggca
     2821 agtccaagct ctcttacaac cagtttaaac aacacatcct gaacctgagc aagtcccagg
     2881 atagaatcag caaaaaaaag aaagagtatc tgctggaaga acgggacatc aacaagttcg
     2941 aggtgcaaaa agagttcatc aatagaaacc tggtggatac ccggtacgcc acaagagagc
     3001 tgacaaacta cctgaaggcc tacttcagcg ccaacaatat gaacgtgaag gtgaaaacga
     3061 tcaacggcag cttcaccgat tacctgcgga aagtgtggaa gtttaagaag gaacggaacc
     3121 acggctacaa gcaccacgcc gaggacgccc tgattatcgc taatgccgat ttcctgttca
     3181 aagagaacaa gaagctgaaa gccgtgaact ctgtgctgga aaaacctgag atcgagagca
     3241 agcagctgga tatccaggtg gatagcgagg ataactacag cgaaatgttc atcatcccta
     3301 agcaggtcca ggacatcaag gacttcagaa acttcaagta cagccacaga gtggacaaga
     3361 agcctaacag acggctgatc aacgatacac tgtacagcac ccggaagaag gacaactcca
     3421 cctacatcgt gcagaccatc aaagatatct atgccaaaga taataccacc ctgaagaagc
     3481 agtttgacaa gtcacccgag aagttcctca tgtaccaaca cgatccgcgg accttcgaga
     3541 agttggaagt gatcatgaag cagtacgcta atgagaagaa tcctctggcc aagtaccacg
     3601 aggaaacagg cgagtacctg accaaataca gcaaaaaaaa caacggccct atcgtgaaaa
     3661 gcctgaagta cattggaaac aagctgggca gacacctaga tgtgacccac cagttcaaga
     3721 gcagcaccaa gaagttggtg aagcggagca tcaagcctta tagattcgac gtctacctga
     3781 ccgacaaggg atataagttc atcaccatca gctacctgga cgtgctgaag aaagacaatt
     3841 actactacat acccgaacag aagtacgaca agctcaaact gggcaaggcc atcgacaaaa
     3901 acgccaagtt tatcgctagc ttctacaaga atgatctgat caagctggac ggcgagatct
     3961 acaagatcat cggcgtgagt agcgacacca gaaacatgat cgaactggat ctgcctgaca
     4021 tcagatacaa agaatactgc gagctgaaca atatcaaggg caaacctaga atcataaaga
     4081 ccatcggcaa aaaggtgaat agcatcgaaa aactgacaac cgacgtgctg ggcaacgtgt
     4141 tcaccaacac ccagtacaca aaacctcagc tgctgttcaa gcgaggaaat tctggcggct
     4201 caaaaagaac cgccgacggc agcgaattcg agcccaagaa gaagaggaaa gtctaatagc
     4261 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     4321 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     4381 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     4441 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggaaa aaaatctcgc
     4501 caacaagttg acgagataaa cacggcattt tgccttgttt tagtagattc tgtttccaga
     4561 gtactaaaac cgaagagcat cgctcttctg gtgtttcgtc ctttccacaa gatatataaa
     4621 gccaagaaat cgaaatactt tcaagttacg gtaagcatat gatagtccat tttaaaacat
     4681 aattttaaaa ctgcaaacta cccaagaaat tattactttc tacgtcacgt attttgtact
     4741 aatatctttg tgtttacagt caaattaatt ccaattatct ctctaacagc cttgtatcgt
     4801 atatgcaaat atgaaggaat catgggaaat aggccctcca cgtgcggacc gagcggccgc
     4861 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     4921 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     4981 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     5041 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     5101 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5161 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5221 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5281 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     5341 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     5401 actcaaccct atctcgggct attcttttga tttataaggg attttgccga tttcggccta
     5461 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     5521 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     5581 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     5641 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     5701 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     5761 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     5821 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5881 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5941 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6001 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     6061 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     6121 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     6181 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     6241 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6301 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6361 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6421 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6481 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6541 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6601 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6661 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6721 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6781 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6841 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6901 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6961 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7021 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7081 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     7141 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7201 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7261 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7321 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7381 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7441 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7501 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7561 acggttcctg gccttttgct ggccttttgc tcacatgt
//
