LOCUS       Exported                6691 bp ds-DNA     circular SYN 04-FEB-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6691)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Feb 4, 2021 from SnapGene Viewer 5.2.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6691
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..130
                     /label=ITR-L
     promoter        145..385
                     /label=U6 promoter
     primer_bind     145..165
                     /label=hU6-F
     misc_feature    395..430
                     /label=CasRx DR (full)
     misc_feature    431..460
                     /label=30nt VEGFA 1 pre-sgRNA
     misc_feature    461..496
                     /label=CasRx DR (full)
     misc_feature    526..781
                     /label=EFS promoter
     misc_feature    788..793
                     /label=Koza
     misc_feature    794..3736
                     /label=CasRx
     CDS             800..820
                     /codon_start=1
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3737..3757
                     /codon_start=1
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3767..3793
                     /codon_start=1
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    3803..3937
                     /label=SV40 polyA
     misc_feature    3954..4094
                     /label=ITR-R
     rep_origin      4169..4624
                     /label=M13 origin
     CDS             5209..5868
                     /codon_start=1
                     /label=AmpR
                     /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     rep_origin      5966..6648
                     /label=ColE1 origin
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgagggc ctatttccca tgattccttc atatttgcat
      181 atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata
      241 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
      301 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
      361 gctttatata tcttgtggaa aggacgaaac accgcaagta aacccctacc aactggtcgg
      421 ggtttgaaac tatgtgctgg ccttggtgag gtttgatccg caagtaaacc cctaccaact
      481 ggtcggggtt tgaaactttt tttgaattct gatgcggtga agctttaggt cttgaaagga
      541 gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
      601 aagttggggg gaggggtcgg caattgatcc ggtgcctaga gaaggtggcg cggggtaaac
      661 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
      721 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
      781 gtctagagcc accatgagcc ccaagaagaa gagaaaggtg gaggccagca tcgaaaaaaa
      841 aaagtccttc gccaagggca tgggcgtgaa gtccacactc gtgtccggct ccaaagtgta
      901 catgacaacc ttcgccgaag gcagcgacgc caggctggaa aagatcgtgg agggcgacag
      961 catcaggagc gtgaatgagg gcgaggcctt cagcgctgaa atggccgata aaaacgccgg
     1021 ctataagatc ggcaacgcca aattcagcca tcctaagggc tacgccgtgg tggctaacaa
     1081 ccctctgtat acaggacccg tccagcagga tatgctcggc ctgaaggaaa ctctggaaaa
     1141 gaggtacttc ggcgagagcg ctgatggcaa tgacaatatt tgtatccagg tgatccataa
     1201 catcctggac attgaaaaaa tcctcgccga atacattacc aacgccgcct acgccgtcaa
     1261 caatatctcc ggcctggata aggacattat tggattcggc aagttctcca cagtgtatac
     1321 ctacgacgaa ttcaaagacc ccgagcacca tagggccgct ttcaacaata acgataagct
     1381 catcaacgcc atcaaggccc agtatgacga gttcgacaac ttcctcgata accccagact
     1441 cggctatttc ggccaggcct ttttcagcaa ggagggcaga aattacatca tcaattacgg
     1501 caacgaatgc tatgacattc tggccctcct gagcggactg aggcactggg tggtccataa
     1561 caacgaagaa gagtccagga tctccaggac ctggctctac aacctcgata agaacctcga
     1621 caacgaatac atctccaccc tcaactacct ctacgacagg atcaccaatg agctgaccaa
     1681 ctccttctcc aagaactccg ccgccaacgt gaactatatt gccgaaactc tgggaatcaa
     1741 ccctgccgaa ttcgccgaac aatatttcag attcagcatt atgaaagagc agaaaaacct
     1801 cggattcaat atcaccaagc tcagggaagt gatgctggac aggaaggata tgtccgagat
     1861 caggaaaaat cataaggtgt tcgactccat caggaccaag gtctacacca tgatggactt
     1921 tgtgatttat aggtattaca tcgaagagga tgccaaggtg gctgccgcca ataagtccct
     1981 ccccgataat gagaagtccc tgagcgagaa ggatatcttt gtgattaacc tgaggggctc
     2041 cttcaacgac gaccagaagg atgccctcta ctacgatgaa gctaatagaa tttggagaaa
     2101 gctcgaaaat atcatgcaca acatcaagga atttagggga aacaagacaa gagagtataa
     2161 gaagaaggac gcccctagac tgcccagaat cctgcccgct ggccgtgatg tttccgcctt
     2221 cagcaaactc atgtatgccc tgaccatgtt cctggatggc aaggagatca acgacctcct
     2281 gaccaccctg attaataaat tcgataacat ccagagcttc ctgaaggtga tgcctctcat
     2341 cggagtcaac gctaagttcg tggaggaata cgcctttttc aaagactccg ccaagatcgc
     2401 cgatgagctg aggctgatca agtccttcgc tagaatggga gaacctattg ccgatgccag
     2461 gagggccatg tatatcgacg ccatccgtat tttaggaacc aacctgtcct atgatgagct
     2521 caaggccctc gccgacacct tttccctgga cgagaacgga aacaagctca agaaaggcaa
     2581 gcacggcatg agaaatttca ttattaataa cgtgatcagc aataaaaggt tccactacct
     2641 gatcagatac ggtgatcctg cccacctcca tgagatcgcc aaaaacgagg ccgtggtgaa
     2701 gttcgtgctc ggcaggatcg ctgacatcca gaaaaaacag ggccagaacg gcaagaacca
     2761 gatcgacagg tactacgaaa cttgtatcgg aaaggataag ggcaagagcg tgagcgaaaa
     2821 ggtggacgct ctcacaaaga tcatcaccgg aatgaactac gaccaattcg acaagaaaag
     2881 gagcgtcatt gaggacaccg gcagggaaaa cgccgagagg gagaagttta aaaagatcat
     2941 cagcctgtac ctcaccgtga tctaccacat cctcaagaat attgtcaata tcaacgccag
     3001 gtacgtcatc ggattccatt gcgtcgagcg tgatgctcaa ctgtacaagg agaaaggcta
     3061 cgacatcaat ctcaagaaac tggaagagaa gggattcagc tccgtcacca agctctgcgc
     3121 tggcattgat gaaactgccc ccgataagag aaaggacgtg gaaaaggaga tggctgaaag
     3181 agccaaggag agcattgaca gcctcgagag cgccaacccc aagctgtatg ccaattacat
     3241 caaatacagc gacgagaaga aagccgagga gttcaccagg cagattaaca gggagaaggc
     3301 caaaaccgcc ctgaacgcct acctgaggaa caccaagtgg aatgtgatca tcagggagga
     3361 cctcctgaga attgacaaca agacatgtac cctgttcaga aacaaggccg tccacctgga
     3421 agtggccagg tatgtccacg cctatatcaa cgacattgcc gaggtcaatt cctacttcca
     3481 actgtaccat tacatcatgc agagaattat catgaatgag aggtacgaga aaagcagcgg
     3541 aaaggtgtcc gagtacttcg acgctgtgaa tgacgagaag aagtacaacg ataggctcct
     3601 gaaactgctg tgtgtgcctt tcggctactg tatccccagg tttaagaacc tgagcatcga
     3661 ggccctgttc gataggaacg aggccgccaa gttcgacaag gagaaaaaga aggtgtccgg
     3721 caattccgga tccggaccta agaaaaagag gaaggtggcg gccgcttacc catacgatgt
     3781 tccagattac gcttaagtcg acaacttgtt tattgcagct tataatggtt acaaataaag
     3841 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3901 gtccaaactc atcaatgtat cttatcatgt ctggatccgg accgagcggc cgcaggaacc
     3961 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     4021 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     4081 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4141 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     4201 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4261 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4321 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4381 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4441 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4501 cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4561 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     4621 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     4681 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4741 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4801 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4861 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4921 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4981 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5041 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5101 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5161 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5221 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5281 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5341 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5401 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5461 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5521 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5581 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5641 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5701 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5761 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5821 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5881 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5941 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6001 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6061 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6121 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6181 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6241 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6301 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6361 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6421 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6481 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6541 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6601 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6661 ctggcctttt gctggccttt tgctcacatg t
//
