LOCUS       py100-sgRNA_hVEGFA        6503 bp ds-DNA     circular     15-FEB-2023
DEFINITION  .
ACCESSION   urn.local...34u-ex9rk9w
VERSION     urn.local...34u-ex9rk9w
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     CDS             join(6188..6503,1..479)
                     /label="Neo(R)"
     polyA_signal    653..783
                     /label="SV40 pA"
     rep_origin      complement(1166..1836)
                     /label="pUC origin"
     CDS             complement(1981..2841)
                     /label="Amp(R)"
     promoter        complement(2842..2940)
                     /label="bla promoter"
     promoter        3147..3395
                     /label="Human U6 Promoter"
     misc_feature    3396..3415
                     /label="VEGFa"
     misc_feature    3416..3491
                     /label="Cas9 scaffold"
     misc_feature    3492..3497
                     /label="U6 terminator"
     promoter        3569..4156
                     /label="CMV promoter"
     misc_feature    4200..4219
                     /label="T7 primer"
     promoter        4200..4216
                     /label="T7 promoter"
     gene            4269..4973
                     /label="mCherry"
     CDS             4974..5000
                     /label="HA"
     polyA_site      5080..5304
                     /label="BGH pA"
     rep_origin      5350..5778
                     /label="f1 origin"
     promoter        5783..6152
                     /label="SV40 early promoter"
ORIGIN      
        1 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
       61 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
      121 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
      181 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
      241 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
      301 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
      361 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
      421 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
      481 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
      541 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
      601 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
      661 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
      721 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
      781 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
      841 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
      901 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
      961 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     1021 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     1081 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     1141 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1201 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1261 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1321 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1381 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1441 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1501 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1561 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1621 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     1681 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1741 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1801 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     1861 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     1921 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1981 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2041 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     2101 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     2161 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     2221 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     2281 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     2341 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     2401 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     2461 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     2521 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     2581 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     2641 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     2701 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     2761 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     2821 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     2881 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     2941 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     3001 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     3061 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     3121 aggcaaggct tgaccgactt aattaagagg gcctatttcc catgattcct tcatatttgc
     3181 atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga
     3241 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
     3301 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct
     3361 tggctttata tatcttgtgg aaaggacgaa acaccggtga gtgagtgtgt gcgtggtttt
     3421 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac
     3481 cgagtcggtg ctttttttaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     3541 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag
     3601 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     3661 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     3721 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     3781 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     3841 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     3901 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     3961 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     4021 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     4081 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     4141 tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt
     4201 aatacgactc actataggga gacccaagct ggctagcgtt taaacttaag cttgccggtg
     4261 ccaccatggt gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct
     4321 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
     4381 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
     4441 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga
     4501 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
     4561 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
     4621 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
     4681 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg
     4741 acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
     4801 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     4861 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     4921 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtacccat
     4981 acgatgttcc agattacgct taagaattct gcagatatcc agcacagtgg cggccgctcg
     5041 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     5101 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     5161 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     5221 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     5281 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     5341 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     5401 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     5461 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5521 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     5581 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5641 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     5701 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     5761 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     5821 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     5881 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     5941 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     6001 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     6061 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     6121 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     6181 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     6241 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     6301 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     6361 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     6421 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     6481 ggggcaggat ctcctgtcat ctc
//
