LOCUS       62056_2540        5998 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 5998)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5998
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        12..315
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        316..519
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     protein_bind    566..590
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             636..656
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             675..1703
                     /codon_start=1
                     /label=Cre
                     /note="site-specific recombinase"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     CDS             1722..1769
                     /codon_start=1
                     /label=nucleoplasmin NLS
                     /note="bipartite nuclear localization signal from
                     nucleoplasmin"
                     /translation="KRPAATKKAGQAKKKK"
     protein_bind    complement(1798..1822)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1838..2426
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    complement(2469..2590)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3118..3706)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3880..4737)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4738..4842)
                     /label=AmpR promoter
     rep_origin      complement(4868..5323)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     protein_bind    5575..5595
                     /label=attB4
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     promoter        5627..5867
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        5877..5912
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
ORIGIN
        1 tcgacggtac ccgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
       61 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      121 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      181 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      241 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      301 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      361 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      421 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      481 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      541 cccgggtaac tgatcataat tcgacccaag tttgtacaaa aaagcaggct gattaccgga
      601 gaattccaat tggcggccgc accggtgcca ccatgccaaa gaagaagcgg aaggtcggtg
      661 gcggctcacc cgggatgtcc aatttactga ccgtacacca aaatttgcct gcattaccgg
      721 tcgatgcaac gagtgatgag gttcgcaaga acctgatgga catgttcagg gatcgccagg
      781 cgttttctga gcatacctgg aaaatgcttc tgtccgtttg ccggtcgtgg gcggcatggt
      841 gcaagttgaa taaccggaaa tggtttcccg cagaacctga agatgttcgc gattatcttc
      901 tatatcttca ggcgcgcggt ctggcagtaa aaactatcca gcaacatttg ggccagctaa
      961 acatgcttca tcgtcggtcc gggctgccac gaccaagtga cagcaatgct gtttcactgg
     1021 ttatgcggcg gatccgaaaa gaaaacgttg atgccggtga acgtgcaaaa caggctctag
     1081 cgttcgaacg cactgatttc gaccaggttc gttcactcat ggaaaatagc gatcgctgcc
     1141 aggatatacg taatctggca tttctgggga ttgcttataa caccctgtta cgtatagccg
     1201 aaattgccag gatcagggtt aaagatatct cacgtactga cggtgggaga atgttaatcc
     1261 atattggcag aacgaaaacg ctggttagca ccgcaggtgt agagaaggca cttagcctgg
     1321 gggtaactaa actggtcgag cgatggattt ccgtctctgg tgtagctgat gatccgaata
     1381 actacctgtt ttgccgggtc agaaaaaatg gtgttgccgc gccatctgcc accagccagc
     1441 tatcaactcg cgccctggaa gggatttttg aagcaactca tcgattgatt tacggcgcta
     1501 aggatgactc tggtcagaga tacctggcct ggtctggaca cagtgcccgt gtcggagccg
     1561 cgcgagatat ggcccgcgct ggagtttcaa taccggagat catgcaagct ggtggctgga
     1621 ccaatgtaaa tattgtcatg aactatatcc gtaacctgga tagtgaaaca ggggcaatgg
     1681 tgcgcctgct ggaagatggc gatactagtg gtggcggctc aaagcgtcct gctgctacta
     1741 agaaagctgg tcaagctaag aaaaagaaag gatccaccgg atctagataa ctgatctacc
     1801 cagctttctt gtacaaagtg gctcgagctc tatcgataat caacctctgg attacaaaat
     1861 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     1921 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     1981 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     2041 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     2101 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     2161 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     2221 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     2281 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     2341 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     2401 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cccgggcggc cgcttcgagc
     2461 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2521 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2581 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     2641 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatcttcc
     2701 tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac aaggaacccc
     2761 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     2821 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     2881 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2941 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3001 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3061 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3121 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3181 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3241 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3301 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3361 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3421 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3481 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3541 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3601 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3661 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3721 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3781 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3841 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3901 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3961 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgtga
     4021 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4081 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4141 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4201 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4261 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4321 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4381 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4441 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4501 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4561 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4621 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4681 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4741 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4801 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4861 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     4921 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     4981 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     5041 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     5101 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     5161 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     5221 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     5281 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg
     5341 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctgcgc
     5401 gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc
     5461 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt
     5521 tccttgtagt taatgattaa cccgccatgc cggttcacca attgtatccg atatcaactt
     5581 tgtatagaaa agttggctcc gaatttctcg aggaattcgg caggaagagg gcctatttcc
     5641 catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa
     5701 tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct
     5761 tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac
     5821 ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccgcaag
     5881 taaaccccta ccaactggtc ggggtttgaa acggagaccg atggtctctc aagtaaaccc
     5941 ctaccaactg gtcggggttt gaaacttttt tttcgagctc aagcttcgaa ttctgcag
//
