LOCUS       Exported                6894 bp ds-DNA     circular SYN 26-AUG-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6894)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Sep 16, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6894
                     /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"
     enhancer        217..520
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        521..724
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    768..777
                     /label=Kozak sequence
     CDS             780..800
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             810..3977
                     /codon_start=1
                     /label=Hsp1Cas9
                     /translation="SDKSKILGFDIGITSIGWAYVEGEELIDCGVRIFTKAENPKTKES
                     LALPRREARGVRRRLARRKGRLNAIKQLICKEFGLNYQDYIAQDGELPKAYAGKLINPY
                     ELRSKALREKLEKEELARVILHIAKHRGYGNKHAKETDDAESGKIKKAISNNEKILNEK
                     GYQSAGEYLYQEFFQKERLQEEKKGNNTTKEFKNVRNKAGDYEHCIAQTQMQNELRLIF
                     EKQREFGLNFSEEFKKKVLETAFFQRPLKDFSDKIGDCTFIEGAKRAPKDSLSAIEFVA
                     LGRIINTLKNLQNTSGEVYSRDKINEILQIVLDKGEISYKKLREILDIESSVRFKDSKL
                     DYTKELKEAEKTKLFESKNLQKFKKALGSFEGISREILDSIAKEIMITKDKEKLAKELK
                     KFPLDETQIQNLSNLSFSQTINLSLEALSQILPFMRGEREDKCLRYDEAVEKAGLKEVA
                     KNKSKDDFLPPLENYEPYLANPVVARALSEYRKVLNALLRKYGKMHKIHIEFTREVGKS
                     KEERNKIEKEQKENFAKNQEAQKQCEILGLPINGANILKMKLWREQQESCVYSGKEIAI
                     DDLKNPAILQVDHIYPYSRSFDDSYNNKVLVFSKENQNKLNKTPFEAFGENKDKWEKIL
                     SLASKLPKPKRQRIYNMQFKDKEQGFLTRNINDTGYIARLVSQWTKNCLSFEKLSEDEV
                     SIAGEKGSKVHVEVISGSLTAMLRHYWGLGDKNRDNHLHHAVDAIIIAYSNEKVIKAFS
                     DYKKHREQNKAKFYAKEIAESEYKKQKVFFEPTNHFRQKVLEKIDSIFVSKPPRKRARG
                     ALHEETFYSFERLCNDKEYAKKEKMDYGGKAGIQKAIALGKIRQVGTKIVSNGTMVRVD
                     IFQHKSNGKFYGVPIYTMDFALGVLPNKAVVSGKDKQGVIKDWLEMDSNYEFCFSLFKD
                     DLILVQKKEMEKAELCYYTDFNSNTAGIKFEKHNNLYVNLNENERKLFTSKEVVKGGKK
                     NVWKDKTPKEIEQLEKNRKIIGGFVTIEDLKTFEKYQVSPLGEVQKADFAPRQAITLKS
                     SPKHRINK"
     primer_bind     3929..3949
                     /label=TS6-correct-gibson-F
     CDS             3981..4001
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4049..4130
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   4157..4297
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4372..4827
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5109..5213
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5214..6074
                     /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      6245..6833
                     /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 cgcgtctagt tattaatagt aatcaattac
      181 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      241 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      301 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      361 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      421 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      481 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      541 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      601 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      661 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      721 agctggttta gtgaaccgtc agatccgcta gagatccgcg gccgcgggcc accatggccc
      781 cgaagaaaaa gcgcaaggtc gaagcgtcca gcgacaaatc caagatcctg ggattcgaca
      841 tcggcatcac cagcatcggc tgggcctacg tggaaggcga agagctgatc gactgcggcg
      901 tccggatctt taccaaggcc gagaatccca agaccaaaga atccctggcc ctgcctagac
      961 gggaagctag aggcgtgcgg cggagactgg cccgaagaaa aggcagactg aatgccatca
     1021 agcaactgat ctgcaaggaa ttcggcctga actaccaaga ctacatcgct caggacggcg
     1081 agctgcccaa agcctacgcc gggaagctga ttaaccctta tgagctgaga tctaaggccc
     1141 ttagagagaa gctggaaaag gaagagctgg cccgagtgat tctgcacatc gccaagcacc
     1201 ggggctacgg aaacaagcat gccaaggaaa ccgacgatgc cgagagcggc aagatcaaga
     1261 aagcaattag caacaatgag aagatcctga atgagaaggg atatcagagc gccggagagt
     1321 acctgtacca ggaatttttc cagaaagaaa gactgcagga ggaaaagaag ggcaacaaca
     1381 cgaccaaaga attcaaaaat gtgaggaaca aagctggcga ctacgagcac tgtatcgctc
     1441 agacccagat gcagaacgag ttgaggctta ttttcgagaa gcagcgagag tttggcctga
     1501 acttcagcga ggaattcaag aagaaagtgc tggagacggc cttttttcag agacctctga
     1561 aagacttcag cgataagatc ggcgattgca ccttcatcga gggcgccaag cgggccccta
     1621 aggacagcct gagcgccatc gagttcgtgg ccctgggaag aatcatcaac accctgaaaa
     1681 atctgcagaa cacaagcggc gaggtgtaca gcagagataa gatcaacgag atcctgcaaa
     1741 tcgtgctgga taagggcgag atcagctaca agaaactgag agaaatcctg gacatcgaga
     1801 gcagcgtgcg tttcaaggac agcaaactgg attacaccaa agaactgaag gaagccgaga
     1861 aaaccaagct gttcgagagc aagaacctcc agaaattcaa gaaagctctg ggcagcttcg
     1921 aaggcatctc cagagagatc ttggacagta tcgccaagga gatcatgatc acaaaggata
     1981 aagagaagct ggccaaggag ctgaagaagt tccctctgga tgaaacccag atccagaacc
     2041 tttctaactt gtccttcagc caaacaatca acctgtcact ggaagccctg tctcagatcc
     2101 tgcccttcat gagaggcgag agagaggaca aatgcctgag atacgacgag gctgtggaaa
     2161 aggccggcct gaaggaggtg gccaaaaaca agtctaagga tgacttcctg cctcctctgg
     2221 aaaactacga gccttacctg gccaatcctg tggtggcccg ggccctctct gagtatagaa
     2281 aggtgctgaa cgccctgctg agaaagtacg gcaagatgca caagattcac atcgagttta
     2341 ccagagaagt gggcaagagc aaagaggagc ggaacaaaat cgaaaaagaa cagaaagaga
     2401 atttcgccaa aaaccaggag gcccagaagc agtgcgagat cctgggcctg cctatcaacg
     2461 gcgctaatat cctgaagatg aagctgtggc gggagcagca ggagtcctgc gtgtacagcg
     2521 gcaaggaaat cgccatcgac gacctgaaga accccgccat cctgcaggtg gatcacatct
     2581 atccttactc tagaagcttt gatgattctt ataacaacaa ggtgctggtg ttctccaaag
     2641 aaaaccagaa caaactcaac aagacacctt tcgaggcttt cggcgaaaac aaggacaaat
     2701 gggagaaaat cctgtccctg gcttctaagc ttcctaagcc taagcggcag aggatctaca
     2761 acatgcagtt caaggacaaa gaacaaggct ttctgacccg gaatatcaac gacaccggct
     2821 acatcgctag actggtcagc cagtggacaa agaactgcct gagcttcgag aagctgagcg
     2881 aggatgaggt ttccatcgcc ggcgagaaag gaagcaaagt gcacgtggaa gtgatcagcg
     2941 gctccctgac agccatgctg cggcactact ggggcctggg cgacaagaac cgggacaacc
     3001 acctgcacca cgcggtggac gccatcatca tcgcttatag caacgaaaag gtgatcaagg
     3061 ccttctctga ttacaaaaaa catagagagc agaacaaggc caagttctac gccaaggaga
     3121 tcgccgagag cgagtacaag aagcagaagg ttttcttcga acctaccaac cacttcagac
     3181 aaaaagtgct ggaaaagatt gacagcatct tcgtgagcaa acctccaagg aagagagccc
     3241 ggggagccct gcacgaggaa acattctaca gcttcgagag actgtgcaac gacaaggagt
     3301 acgccaagaa agagaagatg gactacggcg gaaaggctgg catccagaag gctatcgccc
     3361 tgggtaaaat tagacaggtt ggaacaaaga tcgtgtctaa cggcaccatg gtgcgggtgg
     3421 acatcttcca gcacaagagc aacggcaagt tttacggcgt ccccatctac acaatggatt
     3481 ttgccctcgg cgtgctgcct aacaaggctg tggtgtccgg aaaagataag cagggagtga
     3541 tcaaggactg gctggaaatg gacagcaact acgagttttg cttcagtctc ttcaaggacg
     3601 acctgatcct ggtgcagaaa aaggagatgg aaaaagcaga actgtgctac tacaccgatt
     3661 ttaacagcaa cacggccggc atcaaattcg agaagcacaa caacctgtac gtgaacctga
     3721 atgagaatga aagaaagctg ttcaccagca aggaagttgt taagggcggc aagaagaacg
     3781 tgtggaagga caaaacccct aaagaaatcg agcagctgga gaagaacaga aagatcatcg
     3841 gaggcttcgt gaccatcgag gatctgaaaa ccttcgagaa ataccaggtg tctcctctgg
     3901 gagaggtgca gaaggccgac ttcgccccta gacaggccat caccctgaaa agcagcccta
     3961 agcacagaat caacaagagc cccaagaaga agagaaaggt gtagctcgag ctcgagctcg
     4021 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     4081 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacccacgtg
     4141 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     4201 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     4261 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     4321 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     4381 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4441 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4501 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4561 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     4621 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4681 tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta taagggattt
     4741 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4801 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     4861 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4921 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4981 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     5041 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     5101 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5161 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5221 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5281 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5341 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5401 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5461 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5521 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5581 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5641 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5701 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5761 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5821 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5881 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5941 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     6001 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6061 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6121 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6181 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6241 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6301 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6361 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     6421 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6481 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6541 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6601 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6661 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6721 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6781 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6841 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
