LOCUS       62056_2265        4249 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 4249)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4249
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(49..73)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    89..677
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    complement(720..841)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1369..1957)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2131..2988)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2989..3093)
                     /label=AmpR promoter
     rep_origin      complement(3119..3574)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     protein_bind    3826..3846
                     /label=attB4
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     promoter        3878..4118
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        4128..4163
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     misc_feature    4223..4249
                     /label=MCS
                     /note="multiple cloning site"
ORIGIN
        1 tcgacggtac cgcgggcccg ggatccaccg gatctagata actgatctac ccagctttct
       61 tgtacaaagt ggctcgagct ctatcgataa tcaacctctg gattacaaaa tttgtgaaag
      121 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
      181 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
      241 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
      301 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
      361 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
      421 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
      481 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
      541 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
      601 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
      661 ttgggccgcc tccccgcatc gataccgtcg acccgggcgg ccgcttcgag cagacatgat
      721 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
      781 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
      841 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
      901 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatcttc ctagagcatg
      961 gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg
     1021 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     1081 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcatta
     1141 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     1201 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1261 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1321 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1381 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1441 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1501 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1561 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1621 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1681 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1741 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1801 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1861 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1921 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1981 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2041 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2101 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2161 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     2221 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgtg acccacgctc
     2281 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     2341 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     2401 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     2461 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     2521 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     2581 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     2641 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     2701 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     2761 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     2821 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     2881 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     2941 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3001 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3061 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     3121 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     3181 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     3241 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     3301 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     3361 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3421 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     3481 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3541 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac
     3601 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctgcg cgctcgctcg
     3661 ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca
     3721 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttccttgtag
     3781 ttaatgatta acccgccatg ccggttcacc aattgtatcc gatatcaact ttgtatagaa
     3841 aagttggctc cgaatttctc gaggaattcg gcaggaagag ggcctatttc ccatgattcc
     3901 ttcatatttg catatacgat acaaggctgt tagagagata attagaatta atttgactgt
     3961 aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt
     4021 tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta
     4081 tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccgcaa gtaaacccct
     4141 accaactggt cggggtttga aacggagacc gatggtctct caagtaaacc cctaccaact
     4201 ggtcggggtt tgaaactttt ttttcgagct caagcttcga attctgcag
//
