LOCUS       62056_2330        7206 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 7206)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7206
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        170..582
                     /label=cTnT
                     /note="Chicken cardiac troponin T promoter"
     intron          709..841
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             863..883
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             893..3790
                     /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"
     CDS             3800..3826
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     CDS             3827..3847
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     polyA_signal    3863..3997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4004..4244
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        4254..4283
                     /label=RfxCas13d DR30
                     /note="5' processed 30-base direct repeat for an RfxCas13d
                     gRNA (Konermann et al., 2018)"
     misc_RNA        4314..4349
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     repeat_region   4469..4609
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4684..5139
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5421..5525
                     /label=AmpR promoter
     CDS             5526..6383
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6557..7145
                     /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 gcggccgcac gcgtaagctt cgggcccccc ctcgaggtcg ggataaaagc
      181 agtctgggct ttcacatgac agcatctggg gctgcggcag agggtcgggt ccgaagcgct
      241 gccttatcag cgtccccagc cctgggaggt gacagctggc tggcttgtgt cagcccctcg
      301 ggcactcacg tatctccgtc cgacgggttt aaaatagcaa aactctgagg ccacacaata
      361 gcttgggctt atatgggctc ctgtggggga agggggagca cggagggggc cggggccgct
      421 gctgccaaaa tagcagctca caagtgttgc attcctctct gggcgccggg cacattcctg
      481 ctggctctgc ccgccccggg gtgggcgccg gggggacctt aaagcctctg ccccccaagg
      541 agcccttccc agacagccgc cggcacccac cgctccgtgg gacgatcccc gaagctctag
      601 agctttattg cggtagttta tcacagttaa attgctaacg cagtcagtgc ttctgacaca
      661 acagtctcga acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag
      721 gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc
      781 ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca
      841 gggtacgacc ggtgccacca tgcctaaaaa gaaaagaaag gtgggttctg gtatcgaaaa
      901 aaaaaagtcc ttcgccaagg gcatgggcgt gaagtccaca ctcgtgtccg gctccaaagt
      961 gtacatgaca accttcgccg aaggcagcga cgccaggctg gaaaagatcg tggagggcga
     1021 cagcatcagg agcgtgaatg agggcgaggc cttcagcgct gaaatggccg ataaaaacgc
     1081 cggctataag atcggcaacg ccaaattcag ccatcctaag ggctacgccg tggtggctaa
     1141 caaccctctg tatacaggac ccgtccagca ggatatgctc ggcctgaagg aaactctgga
     1201 aaagaggtac ttcggcgaga gcgctgatgg caatgacaat atttgtatcc aggtgatcca
     1261 taacatcctg gacattgaaa aaatcctcgc cgaatacatt accaacgccg cctacgccgt
     1321 caacaatatc tccggcctgg ataaggacat tattggattc ggcaagttct ccacagtgta
     1381 tacctacgac gaattcaaag accccgagca ccatagggcc gctttcaaca ataacgataa
     1441 gctcatcaac gccatcaagg cccagtatga cgagttcgac aacttcctcg ataaccccag
     1501 actcggctat ttcggccagg cctttttcag caaggagggc agaaattaca tcatcaatta
     1561 cggcaacgaa tgctatgaca ttctggccct cctgagcgga ctgaggcact gggtggtcca
     1621 taacaacgaa gaagagtcca ggatctccag gacctggctc tacaacctcg ataagaacct
     1681 cgacaacgaa tacatctcca ccctcaacta cctctacgac aggatcacca atgagctgac
     1741 caactccttc tccaagaact ccgccgccaa cgtgaactat attgccgaaa ctctgggaat
     1801 caaccctgcc gaattcgccg aacaatattt cagattcagc attatgaaag agcagaaaaa
     1861 cctcggattc aatatcacca agctcaggga agtgatgctg gacaggaagg atatgtccga
     1921 gatcaggaaa aatcataagg tgttcgactc catcaggacc aaggtctaca ccatgatgga
     1981 ctttgtgatt tataggtatt acatcgaaga ggatgccaag gtggctgccg ccaataagtc
     2041 cctccccgat aatgagaagt ccctgagcga gaaggatatc tttgtgatta acctgagggg
     2101 ctccttcaac gacgaccaga aggatgccct ctactacgat gaagctaata gaatttggag
     2161 aaagctcgaa aatatcatgc acaacatcaa ggaatttagg ggaaacaaga caagagagta
     2221 taagaagaag gacgccccta gactgcccag aatcctgccc gctggccgtg atgtttccgc
     2281 cttcagcaaa ctcatgtatg ccctgaccat gttcctggat ggcaaggaga tcaacgacct
     2341 cctgaccacc ctgattaata aattcgataa catccagagc ttcctgaagg tgatgcctct
     2401 catcggagtc aacgctaagt tcgtggagga atacgccttt ttcaaagact ccgccaagat
     2461 cgccgatgag ctgaggctga tcaagtcctt cgctagaatg ggagaaccta ttgccgatgc
     2521 caggagggcc atgtatatcg acgccatccg tattttagga accaacctgt cctatgatga
     2581 gctcaaggcc ctcgccgaca ccttttccct ggacgagaac ggaaacaagc tcaagaaagg
     2641 caagcacggc atgagaaatt tcattattaa taacgtgatc agcaataaaa ggttccacta
     2701 cctgatcaga tacggtgatc ctgcccacct ccatgagatc gccaaaaacg aggccgtggt
     2761 gaagttcgtg ctcggcagga tcgctgacat ccagaaaaaa cagggccaga acggcaagaa
     2821 ccagatcgac aggtactacg aaacttgtat cggaaaggat aagggcaaga gcgtgagcga
     2881 aaaggtggac gctctcacaa agatcatcac cggaatgaac tacgaccaat tcgacaagaa
     2941 aaggagcgtc attgaggaca ccggcaggga aaacgccgag agggagaagt ttaaaaagat
     3001 catcagcctg tacctcaccg tgatctacca catcctcaag aatattgtca atatcaacgc
     3061 caggtacgtc atcggattcc attgcgtcga gcgtgatgct caactgtaca aggagaaagg
     3121 ctacgacatc aatctcaaga aactggaaga gaagggattc agctccgtca ccaagctctg
     3181 cgctggcatt gatgaaactg cccccgataa gagaaaggac gtggaaaagg agatggctga
     3241 aagagccaag gagagcattg acagcctcga gagcgccaac cccaagctgt atgccaatta
     3301 catcaaatac agcgacgaga agaaagccga ggagttcacc aggcagatta acagggagaa
     3361 ggccaaaacc gccctgaacg cctacctgag gaacaccaag tggaatgtga tcatcaggga
     3421 ggacctcctg agaattgaca acaagacatg taccctgttc agaaacaagg ccgtccacct
     3481 ggaagtggcc aggtatgtcc acgcctatat caacgacatt gccgaggtca attcctactt
     3541 ccaactgtac cattacatca tgcagagaat tatcatgaat gagaggtacg agaaaagcag
     3601 cggaaaggtg tccgagtact tcgacgctgt gaatgacgag aagaagtaca acgataggct
     3661 cctgaaactg ctgtgtgtgc ctttcggcta ctgtatcccc aggtttaaga acctgagcat
     3721 cgaggccctg ttcgatagga acgaggccgc caagttcgac aaggagaaaa agaaggtgtc
     3781 cggcaattcc ggttctggtt acccatacga cgtaccagat tacgctccca agaagaagcg
     3841 caaggtggga taaagatcct aaaacttgtt tattgcagct tataatggtt acaaataaag
     3901 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3961 gtccaaactc atcaatgtat cttatcatgt ctggatcgcg gccgagggcc tatttcccat
     4021 gattccttca tatttgcata tacgatacaa ggctgttaga gagataattg gaattaattt
     4081 gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg
     4141 gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg
     4201 aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccgaacccct
     4261 accaactggt cggggtttga aacggggaag ctacactgtt gtccaagcca tcacaagtaa
     4321 acccctacca actggtcggg gtttgaaaca caggagctgg agatgccccc tgaggcaccc
     4381 aagtaaaccc ctaccaactg gtcggggttt gaaacttttt ttgaattctg atgcggtggc
     4441 ggccgccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     4501 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     4561 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     4621 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     4681 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4741 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4801 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4861 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     4921 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4981 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     5041 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     5101 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     5161 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     5221 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     5281 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     5341 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5401 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5461 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5521 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5581 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5641 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5701 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5761 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5821 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5881 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5941 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6001 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6061 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6121 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6181 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6241 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6301 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6361 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6421 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6481 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6541 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6601 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6661 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6721 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6781 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6841 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6901 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6961 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7021 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7081 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7141 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7201 acatgt
//
