LOCUS       pBT276_CDA1-BE4max      8901 bp ds-DNA     circular     22-JAN-2019
DEFINITION  .
FEATURES             Location/Qualifiers
     CDS              702..1322
                      /label="CDA1 Genscript codons"
                      /ApEinfo_revcolor=#b7e6d7
                      /ApEinfo_fwdcolor=#b7e6d7
     cloning junction 6187..6190
                      /label="AGCC for BsaI Golden Gate cloning"
                      /ApEinfo_revcolor=#d59687
                      /ApEinfo_fwdcolor=#d59687
     cloning junction 699..701
                      /label="AGT for SapI Golden Gate cloning"
                      /ApEinfo_revcolor=#c7b0e3
                      /ApEinfo_fwdcolor=#c7b0e3
     CDS              642..6143
                      /label="CDA1-BE4max"
                      /ApEinfo_revcolor=#f58a5e
                      /ApEinfo_fwdcolor=#f58a5e
     cloning junction 6174..6182
                      /label="USER junction 27"
                      /ApEinfo_revcolor=#ff9ccd
                      /ApEinfo_fwdcolor=#ff9ccd
     promoter         1..589
                      /label="CMV promoter, full"
                      /ApEinfo_revcolor=#b1ff67
                      /ApEinfo_fwdcolor=#b1ff67
     Promoter         508..589
                      /label="CMV promoter"
                      /ApEinfo_revcolor=#b4abac
                      /ApEinfo_fwdcolor=#b4abac
     misc_feature     627..627
                      /label="TSS"
                      /ApEinfo_revcolor=#d6b295
                      /ApEinfo_fwdcolor=#d6b295
     misc_feature     630..632
                      /label="modest consensus on RRR"
                      /ApEinfo_revcolor=#85dae9
                      /ApEinfo_fwdcolor=#85dae9
     promoter         610..632
                      /label="T7 promoter"
                      /ApEinfo_revcolor=#f58a5e
                      /ApEinfo_fwdcolor=#f58a5e
     NLS              642..698
                      /label="Bipartite SV40 NLS"
                      /ApEinfo_revcolor=#b1ff67
                      /ApEinfo_fwdcolor=#b1ff67
     CDS              1323..1418
                      /label="BE4 linker Genscript codons"
                      /ApEinfo_revcolor=#d6b295
                      /ApEinfo_fwdcolor=#d6b295
     cloning junction 1334..1337
                      /label="TAGC for BsaI Golden Gate cloning"
                      /ApEinfo_revcolor=#9eafd2
                      /ApEinfo_fwdcolor=#9eafd2
     cloning junction 1335..1344
                      /label="USER junction 33"
                      /ApEinfo_revcolor=#b7e6d7
                      /ApEinfo_fwdcolor=#b7e6d7
     cloning junction 1326..1328
                      /label="GGC for SapI Golden Gate cloning"
                      /ApEinfo_revcolor=#75c6a9
                      /ApEinfo_fwdcolor=#75c6a9
     misc_feature     1443..1445
                      /label="D10A"
                      /ApEinfo_revcolor=#d6b295
                      /ApEinfo_fwdcolor=#d6b295
     CDS              1419..5519
                      /label="Cas9, Genscript codons"
                      /ApEinfo_revcolor=#d59687
                      /ApEinfo_fwdcolor=#d59687
     source           5520..6089
                      /label="2xUGI Genscript codons"
                      /ApEinfo_revcolor=#b7e6d7
                      /ApEinfo_fwdcolor=#b7e6d7
     NLS              6090..6119
                      /label="SV40 NLS"
                      /ApEinfo_revcolor=#b1ff67
                      /ApEinfo_fwdcolor=#b1ff67
     misc_feature     6120..6140
                      /label="SV40 NLS"
                      /ApEinfo_revcolor=#d59687
                      /ApEinfo_fwdcolor=#d59687
     Terminator       6192..6419
                      /label="bGH PA terminator"
                      /ApEinfo_revcolor=#c7b0e3
                      /ApEinfo_fwdcolor=#c7b0e3
     misc_feature     6708..6714
                      /label="removed SapI site"
                      /ApEinfo_revcolor=#faac61
                      /ApEinfo_fwdcolor=#faac61
     misc_feature     7544..7554
                      /label="USER jct 25"
                      /ApEinfo_revcolor=#f8d3a9
                      /ApEinfo_fwdcolor=#f8d3a9
     misc_feature     6710..6718
                      /label="USER jct 26"
                      /ApEinfo_revcolor=#faac61
                      /ApEinfo_fwdcolor=#faac61
     Marker           complement(7651..8511)
                      /label="bla beta lactamase (ampicillin/carbenicillin resistance)"
                      /ApEinfo_revcolor=#ffef86
                      /ApEinfo_fwdcolor=#ffef86
     Promoter         complement(6539..6568)
                      /label="lac promoter"
                      /ApEinfo_revcolor=#d6b295
                      /ApEinfo_fwdcolor=#d6b295
     misc_feature     7791..7796
                      /label="removed BsaI site"
                      /ApEinfo_revcolor=#b7e6d7
                      /ApEinfo_fwdcolor=#b7e6d7
     Promoter         complement(8553..8581)
                      /label="amp promoter"
                      /ApEinfo_revcolor=#faac61
                      /ApEinfo_fwdcolor=#faac61
     misc_feature     7514..7514
                      /label="G>A"
                      /ApEinfo_revcolor=#d59687
                      /ApEinfo_fwdcolor=#d59687
     origin of replication 6874..7554
                      /label="pUC origin high copy"
                      /ApEinfo_revcolor=#9eafd2
                      /ApEinfo_fwdcolor=#9eafd2
     misc_feature     7794..7802
                      /label="USER jct 31"
                      /ApEinfo_revcolor=#9eafd2
                      /ApEinfo_fwdcolor=#9eafd2
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagagatc
      601 cgcggccgct aatacgactc actataggga gagccgccac catgaaacgg acagccgacg
      661 gaagcgagtt cgagtcacca aagaagaagc ggaaagtcag taccgacgcc gagtacgtgc
      721 ggatccacga gaagctggat atctacacat tcaagaagca gttctttaac aacaagaagt
      781 ccgtgtctca cagatgctac gtgctgttcg agctgaagcg gagaggagag aggcgcgcct
      841 gtttttgggg ctatgccgtg aacaagccac agtctggaac cgagagggga atccacgcag
      901 agatcttcag catcaggaag gtggaggagt acctgcgcga caaccccggc cagtttacaa
      961 tcaattggta tagctcctgg agcccttgcg ccgattgtgc cgagaagatc ctggagtggt
     1021 acaaccagga gctgaggggc aatggccacA ccctgaagaT ctgggcctgc aagctgtact
     1081 atgagaagaa cgccaggaat cagatcggcc tgtggaacct gcgcgacaat ggcgtgggcc
     1141 tgaacgtgat ggtgtccgag cactatcagt gctgtcgcaa gatctttatc cagtctagcc
     1201 acaatcagct gaacgagaat cggtggctgg agaagacact gaagagagcc gagaagcgga
     1261 gaAgcgagcT gtccatcatg atccaggtga agatcctgca caccacaaag tctcccgccg
     1321 tgtctggcgg atctAgcgga ggaTcctctg gcagcgagac accaggaaca agcgagtcag
     1381 caacaccaga gagcagtggc ggcagcagcg gcggcagcga caagaagtac agcatcggcc
     1441 tggCcatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca
     1501 gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg
     1561 gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca
     1621 gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg
     1681 agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag
     1741 aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc
     1801 acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg
     1861 ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc
     1921 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
     1981 tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca
     2041 aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc
     2101 tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga
     2161 cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg
     2221 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
     2281 tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga
     2341 acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc
     2401 accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag
     2461 agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc
     2521 aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac
     2581 tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca
     2641 gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt
     2701 tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc
     2761 cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga
     2821 gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc
     2881 agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc
     2941 ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat
     3001 acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg
     3061 tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact
     3121 tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg
     3181 cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca
     3241 atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca
     3301 gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga
     3361 agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg
     3421 gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg
     3481 ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc
     3541 agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg
     3601 gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga
     3661 aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga
     3721 ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca
     3781 aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg
     3841 agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg
     3901 acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctgaagg
     3961 acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca
     4021 acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg
     4081 ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga
     4141 gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa
     4201 agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc
     4261 tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg
     4321 atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc
     4381 tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg
     4441 tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa
     4501 tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg
     4561 agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa
     4621 ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca
     4681 tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt
     4741 ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta
     4801 agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag
     4861 tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca
     4921 tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag
     4981 aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg
     5041 gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc
     5101 cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc
     5161 ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga
     5221 tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca
     5281 aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata
     5341 tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca
     5401 ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc
     5461 accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggtgaca
     5521 gcggcgggag cggcgggagc ggggggagca ctaatctgag cgacatcatt gagaaggaga
     5581 ctgggaaaca gctggtcatt caggagtcca tcctgatgct gcctgaggag gtggaggaag
     5641 tgatcggcaa caagccagag tctgacatcc tggtgcacac cgcctacgac gagtccacag
     5701 atgagaatgt gatgctgctg acctctgacg cccccgagta taagccttgg gccctggtca
     5761 tccaggattc taacggcgag aataagatca agatgctgag cggaggatcc ggaggatctg
     5821 gaggcagcac caacctgtct gacatcatcg agaaggagac aggcaagcag ctggtcatcc
     5881 aggagagcat cctgatgctg cccgaagaag tcgaagaagt gatcggaaac aagcctgaga
     5941 gcgatatcct ggtccatacc gcctacgacg agagtaccga cgaaaatgtg atgctgctga
     6001 catccgacgc cccagagtat aagccctggg ctctggtcat ccaggattcc aacggagaga
     6061 acaaaatcaa aatgctgtct ggcggctcaa aaagaaccgc cgacggcagc gaattcgagc
     6121 ccaagaagaa gaggaaagtc taaccggtca tcatcaccat caccattgag tttAaacccg
     6181 cTgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     6241 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     6301 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     6361 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     6421 ttctgaggcg gaaagaacca gctggggctc gataccgtcg acctctagct agagcttggc
     6481 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6541 catacgagcc ggaagcataa agtgtaaagc ctagggtgcc taatgagtga gctaactcac
     6601 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6661 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcA cttccgcTtc
     6721 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6781 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6841 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6901 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6961 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7021 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7081 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7141 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7201 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7261 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7321 ctacactaga agAacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7381 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7441 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7501 tacggggtct gacActcagt ggaacgaaaa ctcacgttaa gggAttttgg tcaTgagatt
     7561 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7621 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7681 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7741 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gGgAcccacg
     7801 cTcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7861 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7921 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7981 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8041 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8101 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8161 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8221 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8281 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8341 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8401 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8461 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8521 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8581 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8641 tgacgtcgac ggatcgggag atcgatctcc cgatccccta gggtcgactc tcagtacaat
     8701 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     8761 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     8821 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
     8881 gcgttgacat tgattattga c
//