LOCUS       Exported                6819 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 6819)
  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..6819
                     /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..3902
                     /codon_start=1
                     /label=CcuCas9
                     /translation="KILGFDIGISSIGWAFVENEELRDCGVRIFTKAENPENGESLALP
                     RREARGVRRRLARRKARLNGVKKLLCQEFGLKLNDYFSNDGELPKAYQMSKNTKSVYEL
                     RTLALSCKLEFKDLARVFLHIAKHRGYGNKHAKISQDSKDIGQIKKALKHNEEELKNYR
                     SVGEYLYKNFYQKVRNFEEIKTKNKNNQSTFEFKNVRNKSGNYKQCVSQEWLKNELKLI
                     FEKQRSFGVKLSKDFENELLDKIFFQRELKDFSDKIGFCIYFEGKKRASKESLSAKEFI
                     ALTRMINTFKNLEQQNRKNQSGEIYSQEIIKEILNIILDKGMMKYKELRTLLKLDKSIQ
                     FKDSKLSYEKGIEKAEETKFIDFKELRNFKKALGGSFEGLERKHIDEIAHKITLIKSKE
                     KLKIALENYIKEHKLEQRLNECLGNLSELDFSGHIDLSLEALEQILPFMREGFRYDEAV
                     KKAGLREIRKDYLKKDFLPPLKDFENLANPVVVRALAEYRKVLNALLKKYGKMHKIHIE
                     LTREIGLNHKERQNYIEEQKNNFNFNEEARKKCEEIGLDAEKYLLKMKLFLEQDEFCIY
                     SGEKIKIEHLRDAKMLEIDHIYPYSRSFDDSYLNKVLVFSKENQNKGNKTPFEAFGNDK
                     EKWDKIKGLTQGLKKLPSKKKKRILNLYFNDKEAGFISRNLNDTAYIARFSANYTKYFL
                     DFLPLSENEVSISGEKGSKVHVEAISGRLTAMLRHYWGLGQKDRNNHLHHAVDAVIVAY
                     TNAKIIKNFADFKKFREQNKAKIYAKEFAQTTDKKIFKPCENFRNKILMKIDEIFVSKP
                     PRKRARGALHKESFMSFDDKKLLKSYGGREGVKKALSLGKIRKIGTKIVENKNMVRVDI
                     FKDKKGKFYGVPIYTMDFALGILPNKAVVSGEDKKTREIKDWKEMDEKYEFCFSLFKDD
                     LIKIQKKDMKEPKFCYYKNFDVSNASICIAKHDNKFENRDEDEKILYSKATYKKVEARG
                     IGIQNLKILEKWQVSPLGEKREFEFEPRKNISLKTTNRKNVF"
     CDS             3906..3926
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3974..4055
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   4082..4222
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4297..4752
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5034..5138
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5139..5999
                     /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      6170..6758
                     /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 agatcttggg ctttgatatc ggaatctcca
      841 gcatcggatg ggccttcgtg gaaaatgagg aactgcggga ctgcggcgtc cggatcttca
      901 ccaaggccga gaaccccgag aacggcgaga gccttgccct gcctagacgg gaagctagag
      961 gcgtgagaag aaggctggct agaagaaagg cccggctgaa cggcgtgaag aaactgctgt
     1021 gccaggagtt cggcctcaag ctgaacgact acttcagcaa cgacggcgag ctgcctaagg
     1081 cttatcagat gagcaagaat accaagagcg tgtacgagct gagaaccctg gccctgagct
     1141 gcaagctgga gttcaaggac ctggctagag tgtttctgca catcgccaaa catagaggct
     1201 acgggaacaa gcacgccaag atcagccaag acagcaagga tatcggccaa atcaagaagg
     1261 ccctgaaaca taatgaagag gagctgaaga attaccggag tgtgggcgag tacctgtaca
     1321 agaacttcta ccagaaggtg cggaatttcg aagaaatcaa gacaaagaac aagaacaacc
     1381 agagcacctt cgagttcaag aacgtgcgaa acaagtctgg caactacaag caatgtgtga
     1441 gccaggagtg gctgaaaaac gagctgaagc tgatcttcga gaagcagaga tctttcggcg
     1501 tgaaactgtc caaggatttc gagaacgagc tgctggacaa gatctttttc cagcgggagc
     1561 taaaggattt tagcgacaaa atcggctttt gcatatactt tgaaggcaag aaaagagcca
     1621 gcaaggaaag cctcagcgcc aaggaattca tcgcgttgac aagaatgatc aacacattca
     1681 agaaccttga acagcagaac cggaagaacc agtctgggga gatctactcc caggagatca
     1741 tcaaggagat tttgaacatc atcctggaca agggcatgat gaagtacaaa gaactgagga
     1801 ccctgctgaa gcttgataaa tccatccagt tcaaggatag caagcttagt tacgagaagg
     1861 ggatcgagaa ggccgaggaa accaaattca ttgatttcaa ggaactgcgc aacttcaaga
     1921 aggctcttgg cggatctttc gaaggcctgg agagaaagca tattgacgaa attgcccaca
     1981 agataacact gatcaagtcc aaggagaagc tgaaaattgc tctggagaac tacatcaagg
     2041 aacacaagct ggaacagaga cttaacgagt gcctgggaaa cctgtctgag ctggatttca
     2101 gcggccacat cgacctgtct ctggaggccc tggagcagat cctgcctttt atgagagagg
     2161 gcttcagata cgacgaggcc gtgaagaagg ccggcctgag agagatccgg aaagattacc
     2221 tgaagaagga cttcctgcca cctctgaagg actttgagaa cctggctaat cctgtggtgg
     2281 tgagagccct cgctgaatac agaaaagtgc tgaacgccct gctgaagaag tacggaaaaa
     2341 tgcacaagat tcacattgag ctgacaagag agatcgggct gaaccacaaa gaaagacaga
     2401 actatatcga ggaacaaaag aacaacttta actttaacga ggaagccaga aagaagtgcg
     2461 aggagatcgg cttggatgcc gagaagtacc tgctgaagat gaaactgttc ctggagcagg
     2521 acgagttctg tatctacagc ggagagaaga tcaagatcga gcacctgaga gatgccaaga
     2581 tgctggaaat cgaccacatc tacccctaca gcagaagctt cgatgacagc tacctgaaca
     2641 aggtgctggt gttctccaag gaaaaccaga acaaaggaaa caagacccct ttcgaggctt
     2701 ttggcaatga caaagagaag tgggacaaga tcaaaggcct gacccagggc ctgaagaagc
     2761 tgcccagcaa gaaaaagaaa agaatcctga acctctactt caatgacaag gaagctggct
     2821 tcatcagcag aaacctgaac gacaccgcct acatcgccag attctccgcc aactacacaa
     2881 aatacttcct ggacttcctg cccctgagcg agaacgaagt gtccatatcc ggagagaaag
     2941 gctctaaggt gcacgtggaa gccatcagcg gcagactgac agccatgctg agacactact
     3001 ggggcctggg ccagaaggac agaaacaacc acctgcacca cgccgtggac gcagtgatcg
     3061 tggcctacac caacgccaag atcatcaaga acttcgccga cttcaagaag ttccgggagc
     3121 aaaacaaggc caagatctac gccaaggaat tcgcccagac caccgacaag aaaatcttta
     3181 aaccctgtga aaatttcaga aacaagatcc tcatgaagat tgacgagatc tttgtgtcca
     3241 aacctccaag aaagcgggcc cggggcgccc tgcacaagga gagcttcatg agcttcgacg
     3301 acaagaagct gctcaaaagc tacggaggaa gagagggcgt caagaaggcc ctgtccctgg
     3361 gcaagatcag aaagatcggt acaaagatag ttgagaacaa gaatatggtg cgggtcgaca
     3421 tcttcaagga caagaagggc aagttctacg gcgtccccat ctatacaatg gatttcgctc
     3481 tgggaatcct gcctaacaaa gccgtggtga gtggagaaga taagaaaacc cgggaaatca
     3541 aggactggaa ggaaatggac gagaagtacg aattctgctt cagcctgttc aaagacgatc
     3601 tgattaaaat tcagaaaaag gatatgaagg aacctaagtt ctgctactac aagaatttcg
     3661 acgtgtccaa cgcttctatc tgcatcgcaa aacacgacaa caagttcgag aatagagacg
     3721 aggacgaaaa aatcctgtac agcaaagcca cctacaagaa ggtggaagcc agaggcatcg
     3781 gcatccagaa tctgaagatc ctggagaagt ggcaggtgag tcctctgggc gaaaagcggg
     3841 aattcgagtt cgagcctcgg aagaatatct cactgaaaac caccaacaga aagaatgtgt
     3901 tcagccccaa gaagaagaga aaggtgtagc tcgagctcga gctcgatgag tttggacaaa
     3961 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     4021 tatttgtaac cattataagc tgcaataaac aagttaaccc acgtgcggac cgagcggccg
     4081 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     4141 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     4201 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     4261 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     4321 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4381 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4441 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4501 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4561 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4621 cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct
     4681 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     4741 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4801 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4861 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4921 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     4981 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5041 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5101 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5161 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5221 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5281 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5341 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5401 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5461 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5521 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5581 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5641 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5701 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5761 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5821 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5881 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5941 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6001 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6061 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6121 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6181 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6241 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6301 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6361 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6421 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6481 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6541 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6601 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6661 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6721 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6781 tacggttcct ggccttttgc tggccttttg ctcacatgt
//
