LOCUS       62056_17135        7559 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7559)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     enhancer        221..524
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        525..727
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     intron          876..1254
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and
                     human beta-globin intron 2"
     CDS             1340..1360
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             1370..4267
                     /codon_start=1
                     /label=RfxCas13d
                     /note="RNA-targeting Type VI CRISPR protein from
                     Ruminococcus flavefaciens strain XPD3002 (Konermann et al.,
                     2018)"
                     /translation="IEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDSI
                     RSVNEGEAFSAEMADKNAGYKIGNAKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLEK
                     RYFGESADGNDNICIQVIHNILDIEKILAEYITNAAYAVNNISGLDKDIIGFGKFSTVY
                     TYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFSKEGRNYIIN
                     YGNECYDILALLSGLRHWVVHNNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITNE
                     LTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRKD
                     MSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFVI
                     NLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPAG
                     RDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAFF
                     KDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDEN
                     GNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQK
                     KQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGREN
                     AEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLEE
                     KGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKKA
                     EEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFRNKAVHLEVARYVHA
                     YINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCVP
                     FGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNS"
     polyA_signal    4306..4782
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   4822..4962
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      5037..5492
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5774..5878
                     /label=AmpR promoter
     CDS             5879..6736
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6910..7498
                     /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 cgcgtggagc tagttattaa tagtaatcaa
      181 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      241 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      301 ttcccatagt aacgtcaata gggactttcc attgacgtca atgggtggag tatttacggt
      361 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      421 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      481 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      541 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      601 ttgacgtcaa tgggagtttg ttttgcacca aaatcaacgg gactttccaa aatgtcgtaa
      661 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      721 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
      781 ccatagaaga caccgggacc gatccagcct ccgcggattc gaatcccggc cgggaacggt
      841 gcattggaac gcggattccc cgtgccaaga gtgacgtaag taccgcctat agagtctata
      901 ggcccacaaa aaatgctttc ttcttttaat atactttttt gtttatctta tttctaatac
      961 tttccctaat ctctttcttt cagggcaata atgatacaat gtatcatgcc tctttgcacc
     1021 attctaaaga ataacagtga taatttctgg gttaaggcaa tagcaatatt tctgcatata
     1081 aatatttctg catataaatt gtaactgatg taagaggttt catattgcta atagcagcta
     1141 caatccagct accattctgc ttttatttta tggttgggat aaggctggat tattctgagt
     1201 ccaagctagg cccttttgct aatcatgttc atacctctta tcttcctccc acagctcctg
     1261 ggcaacgtgc tggtctgtgt gctggcccat cactttggca aagaattggg attcgaacat
     1321 cgattgaatt gccaccatgc ccaagaagaa gagaaaggtg gaggccagca tcgaaaaaaa
     1381 aaagtccttc gccaagggca tgggcgtgaa gtccacactc gtgtccggct ccaaagtgta
     1441 catgacaacc ttcgccgaag gcagcgacgc caggctggaa aagatcgtgg agggcgacag
     1501 catcaggagc gtgaatgagg gcgaggcctt cagcgctgaa atggccgata aaaacgccgg
     1561 ctataagatc ggcaacgcca aattcagcca tcctaagggc tacgccgtgg tggctaacaa
     1621 ccctctgtat acaggacccg tccagcagga tatgctcggc ctgaaggaaa ctctggaaaa
     1681 gaggtacttc ggcgagagcg ctgatggcaa tgacaatatt tgtatccagg tgatccataa
     1741 catcctggac attgaaaaaa tcctcgccga atacattacc aacgccgcct acgccgtcaa
     1801 caatatctcc ggcctggata aggacattat tggattcggc aagttctcca cagtgtatac
     1861 ctacgacgaa ttcaaagacc ccgagcacca tagggccgct ttcaacaata acgataagct
     1921 catcaacgcc atcaaggccc agtatgacga gttcgacaac ttcctcgata accccagact
     1981 cggctatttc ggccaggcct ttttcagcaa ggagggcaga aattacatca tcaattacgg
     2041 caacgaatgc tatgacattc tggccctcct gagcggactg aggcactggg tggtccataa
     2101 caacgaagaa gagtccagga tctccaggac ctggctctac aacctcgata agaacctcga
     2161 caacgaatac atctccaccc tcaactacct ctacgacagg atcaccaatg agctgaccaa
     2221 ctccttctcc aagaactccg ccgccaacgt gaactatatt gccgaaactc tgggaatcaa
     2281 ccctgccgaa ttcgccgaac aatatttcag attcagcatt atgaaagagc agaaaaacct
     2341 cggattcaat atcaccaagc tcagggaagt gatgctggac aggaaggata tgtccgagat
     2401 caggaaaaat cataaggtgt tcgactccat caggaccaag gtctacacca tgatggactt
     2461 tgtgatttat aggtattaca tcgaagagga tgccaaggtg gctgccgcca ataagtccct
     2521 ccccgataat gagaagtccc tgagcgagaa ggatatcttt gtgattaacc tgaggggctc
     2581 cttcaacgac gaccagaagg atgccctcta ctacgatgaa gctaatagaa tttggagaaa
     2641 gctcgaaaat atcatgcaca acatcaagga atttagggga aacaagacaa gagagtataa
     2701 gaagaaggac gcccctagac tgcccagaat cctgcccgct ggccgtgatg tttccgcctt
     2761 cagcaaactc atgtatgccc tgaccatgtt cctggatggc aaggagatca acgacctcct
     2821 gaccaccctg attaataaat tcgataacat ccagagcttc ctgaaggtga tgcctctcat
     2881 cggagtcaac gctaagttcg tggaggaata cgcctttttc aaagactccg ccaagatcgc
     2941 cgatgagctg aggctgatca agtccttcgc tagaatggga gaacctattg ccgatgccag
     3001 gagggccatg tatatcgacg ccatccgtat tttaggaacc aacctgtcct atgatgagct
     3061 caaggccctc gccgacacct tttccctgga cgagaacgga aacaagctca agaaaggcaa
     3121 gcacggcatg agaaatttca ttattaataa cgtgatcagc aataaaaggt tccactacct
     3181 gatcagatac ggtgatcctg cccacctcca tgagatcgcc aaaaacgagg ccgtggtgaa
     3241 gttcgtgctc ggcaggatcg ctgacatcca gaaaaaacag ggccagaacg gcaagaacca
     3301 gatcgacagg tactacgaaa cttgtatcgg aaaggataag ggcaagagcg tgagcgaaaa
     3361 ggtggacgct ctcacaaaga tcatcaccgg aatgaactac gaccaattcg acaagaaaag
     3421 gagcgtcatt gaggacaccg gcagggaaaa cgccgagagg gagaagttta aaaagatcat
     3481 cagcctgtac ctcaccgtga tctaccacat cctcaagaat attgtcaata tcaacgccag
     3541 gtacgtcatc ggattccatt gcgtcgagcg tgatgctcaa ctgtacaagg agaaaggcta
     3601 cgacatcaat ctcaagaaac tggaagagaa gggattcagc tccgtcacca agctctgcgc
     3661 tggcattgat gaaactgccc ccgataagag aaaggacgtg gaaaaggaga tggctgaaag
     3721 agccaaggag agcattgaca gcctcgagag cgccaacccc aagctgtatg ccaattacat
     3781 caaatacagc gacgagaaga aagccgagga gttcaccagg cagattaaca gggagaaggc
     3841 caaaaccgcc ctgaacgcct acctgaggaa caccaagtgg aatgtgatca tcagggagga
     3901 cctcctgaga attgacaaca agacatgtac cctgttcaga aacaaggccg tccacctgga
     3961 agtggccagg tatgtccacg cctatatcaa cgacattgcc gaggtcaatt cctacttcca
     4021 actgtaccat tacatcatgc agagaattat catgaatgag aggtacgaga aaagcagcgg
     4081 aaaggtgtcc gagtacttcg acgctgtgaa tgacgagaag aagtacaacg ataggctcct
     4141 gaaactgctg tgtgtgcctt tcggctactg tatccccagg tttaagaacc tgagcatcga
     4201 ggccctgttc gataggaacg aggccgccaa gttcgacaag gagaaaaaga aggtgtccgg
     4261 caattcctga agcttgcctc gagcagcgct gctcgagaga tctacgggtg gcatccctgt
     4321 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt
     4381 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg
     4441 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc
     4501 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc
     4561 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca
     4621 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
     4681 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
     4741 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc
     4801 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     4861 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     4921 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     4981 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     5041 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5101 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5161 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5221 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     5281 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     5341 cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg
     5401 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5461 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     5521 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     5581 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     5641 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     5701 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     5761 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5821 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     5881 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     5941 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     6001 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6061 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6121 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6181 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     6241 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     6301 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6361 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6421 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6481 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6541 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6601 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     6661 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     6721 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     6781 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6841 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6901 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6961 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7021 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     7081 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7141 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7201 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7261 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     7321 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7381 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7441 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7501 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
