LOCUS       pU6-sgTREX1.g1-Cas9-T2A 9282 bp ds-DNA     circular     08-DEC-2020
DEFINITION  .
FEATURES             Location/Qualifiers
     rep_origin      6760..7215
                     /label="f1 ori"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    256..272
                     /label="TREX1.g1 (JM273+274)"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     CDS             7602..8462
                     /label="AmpR"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     CDS             5528..5581
                     /label="T2A"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     promoter        7..247
                     /label="U6 promoter"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     CDS             1376..5476
                     /label="Cas9"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     promoter        7497..7601
                     /label="AmpR promoter"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     polyA_signal    6329..6536
                     /label="bGH poly(A) signal"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_RNA        273..348
                     /label="gRNA scaffold"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     CDS             1259..1324
                     /label="3xFLAG"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     CDS             5588..6298
                     /label="mCherry"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     rep_origin      8633..9221
                     /label="ori"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     enhancer        445..730
                     /label="CMV enhancer"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     CDS             1331..1351
                     /label="SV40 NLS"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     CDS             5477..5524
                     /label="nucleoplasmin NLS"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
ORIGIN
        1 ccccacgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
       61 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
      121 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
      181 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
      241 aaaggacgaa aCACCGAGCC CCCCCACCTC TCgttttaga gctagaaata gcaagttaaa
      301 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt ttttgtttta
      361 gagctagaaa tagcaagtta aaataaggct agtccgtttt tagcgcgtgc gccaattctg
      421 cagacaaatg gctctagagg tacccgttac ataacttacg gtaaatggcc cgcctggctg
      481 accgcccaac gacccccgcc cattgacgtc aatagtaacg ccaataggga ctttccattg
      541 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      601 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattgtgc
      661 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      721 tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc
      781 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg
      841 gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag
      901 gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc
      961 gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc
     1021 gacgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct
     1081 gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta
     1141 attagctgag caagaggtaa gggtttaagg gatggttggt tggtggggta ttaatgttta
     1201 attacctgga gcacctgcct gaaatcactt tttttcaggt tggaccggtg ccaccatgga
     1261 ctataaggac cacgacggag actacaagga tcatgatatt gattacaaag acgatgacga
     1321 taagatggcc ccaaagaaga agcggaaggt cggtatccac ggagtcccag cagccgacaa
     1381 gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg tgatcaccga
     1441 cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat
     1501 caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg
     1561 gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca
     1621 agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga
     1681 gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt
     1741 ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt
     1801 ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa
     1861 gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa
     1921 gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc
     1981 cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga
     2041 aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc
     2101 cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa
     2161 actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg
     2221 cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag
     2281 cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa
     2341 gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct
     2401 gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat
     2461 tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat
     2521 ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg
     2581 gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct
     2641 gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat
     2701 cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc
     2761 ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga
     2821 caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc
     2881 caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga
     2941 gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga
     3001 gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca
     3061 gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt
     3121 ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga
     3181 caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct
     3241 gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt
     3301 cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag
     3361 ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct
     3421 gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac
     3481 ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca
     3541 cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt
     3601 ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat
     3661 ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg
     3721 gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa
     3781 cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta
     3841 cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc
     3901 tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa
     3961 ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg
     4021 gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc
     4081 cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga
     4141 aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta
     4201 cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt
     4261 gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca
     4321 cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa
     4381 gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc
     4441 caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat
     4501 gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat
     4561 cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt
     4621 gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg
     4681 cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa
     4741 gaaggactgg gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt
     4801 gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct
     4861 gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga
     4921 agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct
     4981 gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg
     5041 aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga
     5101 gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa
     5161 gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc
     5221 cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag
     5281 agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc
     5341 cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct
     5401 ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc
     5461 tcagctggga ggcgacaaaa ggccggcggc cacgaaaaag gccggccagg caaaaaagaa
     5521 aaagcttgag ggcagaggaa gtctgctaac atgcggtgac gtggaggaga atcccggccc
     5581 tgctagcatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
     5641 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     5701 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
     5761 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     5821 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     5881 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     5941 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctcagacgg
     6001 ccccgtaatg cagaagaaaa ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     6061 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     6121 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     6181 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     6241 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaaga
     6301 attcctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     6361 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     6421 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     6481 gtggggcagg acagcaaggg ggaggattgg gaagagaata gcaggcatgc tggggagcgg
     6541 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     6601 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     6661 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     6721 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     6781 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     6841 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     6901 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     6961 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     7021 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     7081 caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     7141 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     7201 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     7261 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     7321 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     7381 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     7441 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     7501 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     7561 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     7621 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     7681 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     7741 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     7801 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     7861 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     7921 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     7981 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     8041 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     8101 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     8161 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     8221 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     8281 ggtttattgc tgataaatct ggagccggtg agcgtggaag ccgcggtatc attgcagcac
     8341 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     8401 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     8461 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     8521 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     8581 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     8641 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     8701 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     8761 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     8821 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     8881 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     8941 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     9001 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     9061 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     9121 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     9181 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     9241 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//