LOCUS       pFRT_TO_FLAGHA_S        9245 bp ds-DNA     circular     29-JAN-2019
DEFINITION  .
ACCESSION   
VERSION     
SOURCE      .
  ORGANISM  .
COMMENT     Insert from pDONR223 SCAF8 isoform C Ex1+8 CRISPR resistant 654 to
            4637
COMMENT     Backbone from pFRT_TO_FLAGHA_DEST 2749 to 1066
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     misc_feature    join(1704..1705,1707..1708,1710..1711,1713..1714,
                     1716..1717,1719..1720,1722..1723,1725..1726)
                     /locus_tag="SCAf8_Ex8_gRNA"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_recomb     5711..5758
                     /vntifkey="86"
                     /locus_tag="FRT"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             990..1058
                     /vntifkey="4"
                     /locus_tag="FLAG/HA"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    join(1740..1740,1737..1737)
                     /locus_tag="SCAf8_Ex8_gRNA(1)"
                     /ApEinfo_label="SCAf8_Ex8_gRNA"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             5766..6786
                     /vntifkey="4"
                     /locus_tag="Hygromycin"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1059..1066
                     /vntifkey="21"
                     /locus_tag="attR1"
                     /ApEinfo_fwdcolor="#80ff00"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2113..2113
                     /locus_tag="Mut correction from ATT(I) to AAT (N)"
                     /ApEinfo_fwdcolor="#ff0080"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    complement(8249..9109)
                     /vntifkey="21"
                     /locus_tag="Ampicillin"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    5051..5065
                     /vntifkey="21"
                     /locus_tag="attR2"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2755..2760
                     /locus_tag="Destroyed XmaI site"
                     /ApEinfo_fwdcolor="#cccccc"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     rep_origin      complement(7431..8104)
                     /vntifkey="33"
                     /locus_tag="pUC\ori"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    complement(1067..1067)
                     /SerialCloner_Color=&hDC143C
                     /SerialCloner_Show=False
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=label="attL1" - complement(569..652) \ 
                     /locus_tag="attL1"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     polyA_signal    5203..5427
                     /vntifkey="25"
                     /locus_tag="BGH"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    join(1089..1114,1286..1668)
                     /locus_tag="SCAF8 ORF"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        232..958
                     /dnas_title="CMV"
                     /vntifkey="29"
                     /locus_tag="CMV"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2803..5039
                     /locus_tag="SCAF8 ORF(1)"
                     /ApEinfo_label="SCAF8 ORF"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     polyA_signal    6918..7048
                     /vntifkey="25"
                     /locus_tag="SV40\early"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    990..1058
                     /locus_tag="FLAG/HA tag"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggaacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtacca tggactacaa ggacgacgat gacaagtacc
     1021 cttatgacgt gcccgattac gctttgaacc agatatcaac aagtttGTAC AAAAAAGCAG
     1081 GCTTCACCat gccgcagccg ctgctgccag cgctTCCTCC TTTAAGCAGC CCGTCAGGGG
     1141 CTGGAAGCTG TGGACCATCA GGAGGGGAGT GCAGTGGCCG CCGCCTCTTC CGCCGCCGGG
     1201 CTCGGGGCCT CCGCAGCGAC AACATGGAGG CCGTGAAGAC CTTCAATAGC GAGTTGTATT
     1261 CCCTGAATGA CTATAAACCA CCCATttcga aagcgaaaat gacccaaatt actaaggcag
     1321 ccatcaaagc tattaagttc tataaacatg tggtacagag tgttgagaag tttattcaga
     1381 aatgtaaacc agaatacaaa gtacctggac tttatgttat tgactccatt gtgcgacaat
     1441 cccgacatca gtttggtcaa gaaaaggatg tgtttgcacc cagatttagt aataacatca
     1501 ttagcacttt ccagaattta tatcgttgcc ctggggatga caagagtaaa atagtgagag
     1561 tactaaactt atggcagaag aataatgtat ttaagagtga gattattcag ccccttttgg
     1621 atatggcagc cgggattccg cctccagttg tcacacctgt tttggccagc actaccactg
     1681 ctatgagcaa tactccagga actccagtca cgccggtcac gcctgccaat gtggttcagg
     1741 gattgccaga cccatgggta tctcagataa caaatacaga tacacttgcg gctgtagctc
     1801 agatcttgca aagtcctcaa ggccagcagc ttcaacaatt aatacaaacc ttacagatac
     1861 aacaacagaa gccccagcct tccattctgc aggccctaga tgctggtctt gttgttcagt
     1921 tgcaagctct tacggcacaa cttacagctg cagctgcagc tgccaacact cttactccct
     1981 tagaacaggg agtctccttt aacaagaagt tgatggatag gtttgatttt ggggaagact
     2041 ctgagcatag tgaagaaccc aaaaaggaaa ttccagcttc acaactttct cacgtttcag
     2101 aatctgtgaa caAttccatt tttcatcaga tagcagaaca actacaacag caaaacctag
     2161 aacatctcag acagcagctc ttggagcagc aacagcctca aaaggccact cctcaggata
     2221 gtcaggaagg aacctttggg tcagagcatt cagcgtcacc atcacaaggg agtagtcagc
     2281 agcattttct tgaacctgaa gtcaatttgg atgattccat agatattcag caacaggata
     2341 tggatataga tgaagggcaa gatggagtgg aagaggaggt ctttgaacaa gaagctaaga
     2401 aagtggcggt tcgctcaaga tcaagaacac attcacgatc tcgttcaaga tcaccaagaa
     2461 aacgaaggtc taggtcacgg tctggctcta gaaagcgtaa acacagaaag cgatcacgct
     2521 cccgctcaag agaaagaaag aggaaatcat cacggtcgta ttcaagtgaa aggagagcca
     2581 gagaaaggga gaaagaacga cagaaaaagg gattacctcc aattagatct aaaacactaa
     2641 gtgtatgtag tactactctc tgggttgggc aagtggacaa gaaggcaaca cagcaagact
     2701 taaccaacct gtttgaagag tttggacaga ttgaatccat taatatgatt cctccgcgag
     2761 gctgtgctta tgtctgcatg gttcatcgac aagatgcatt tcgagctctt cagaaactca
     2821 gttctggatc atataaaatt gggtccaagg tcattaagat cgcttgggct ttaaacaaag
     2881 gtgtaaaaac agaatacaaa caattctggg atgtggatct tggagttaca tatataccat
     2941 gggaaaaagt taaagtggat gacttggaag gttttgcaga aggaggcatg attgatcagg
     3001 agactgtaaa tactgagtgg gaaactgtga aaagctcaga acctgttaaa gagacggtcc
     3061 agacaactca gagcccaact ccagttgaaa aggagacagt ggtcacaacc caggcagagg
     3121 ttttccctcc tcctgttgct atgttgcaga ttccagtggc gccagccgtg cctacagtta
     3181 gtttagtccc accagcattt cctgtgtcga tgccggttcc tcctcctgga ttcagtccaa
     3241 tccctccacc tcctttttta agagcaagtt ttaacccttc acaaccacca cctggtttca
     3301 tgccgcctcc agttccccca cctgttgtgc caccccctac gattccacca gtagtaccaa
     3361 catctttagt gcagccgtca ttatccatga caccggaaac tgtgaaagat gttggatttg
     3421 gtagccttgt tataccaggc ggttctgttg ccagcaatct tgctacttcc gctctgccag
     3481 ctggaaatgt ttttaatgct ccaactaaac aggcagagcc tgaagaaaaa gtacctcatc
     3541 ttatagacca ccagatttct tctggtgaaa acaccagatc agtgattcca aatgatattt
     3601 caagtaatgc tgcaatttta ggaggacagc cgccaaatgt gacaagcaat tctggaattc
     3661 tgggagtcca aagaccaaat gtatcaagta attctgaaat tcttggggtc cggccatcta
     3721 atgtttccag tagttctggg attattgcag cccaaccacc aaatattcta aataactctg
     3781 gaatattggg aatacagcca cccagtgtgt caaatagttc tggacttttg ggagtgctac
     3841 ccccaaatat acctaacaat tctggacttg taggagtaca gccaccaaat gttccaaata
     3901 ctcctggact tctgggaaca cagccaccag ctggacctca aaacttaccc cctttaagta
     3961 tccctaatca aaggatgccc acaatgccaa tgttagacat tcgtccggga ctaataccac
     4021 aggcacctgg gccaagattc cctttaatac agcctggaat tccaccccaa cggggaatcc
     4081 cacccccatc ggtacttgat tcagctcttc atccaccacc ccgtggacct tttcctccag
     4141 gagatatttt tagtcaacca gaaagacctt ttttagctcc tggaagacaa agcgtagaca
     4201 atgttactaa cccagaaaaa aggataccac ttgggaatga taacattcaa caggaaggag
     4261 atagagatta ccggtttcct cctatagaaa ccagggaaag cattagtaga cctccccctg
     4321 tggatgttag agatgtggtt gggcggccta tagatccaag agaaggtcct ggacggcctc
     4381 cactagatgg tagggatcat tttggaagac ctcctgtaga tataagagag aatcttgtga
     4441 ggccaggtat agatcatctt ggtcgaagag accactttgg ctttaatcca gagaagccct
     4501 gggggcatag aggagatttt gatgagagag agcatcgggt tctaccggtc tatggtggtc
     4561 caaaaggctt acatgaagaa agaggtagat ttcggtctgg aaactatcga tttgatccta
     4621 gaagtggtcc ttggaaccga ggatttggac aagaagttca cagagatttt gatgaccgca
     4681 gaagaccctg ggagaggcaa agggataggg atgacagaga ttttgatttc tgcagagaaa
     4741 tgaatggaaa tcgtcttgga cgagacagaa ttcaaaacac ttgggttccc cctcctcatg
     4801 ctcgggtttt tgattatttt gaaggggcca cttctcaacg aaaaggtgat aatgtgcctc
     4861 aggttaatgg tgaaaataca gagagacatg ctcagccacc acctatacca gtacagaatg
     4921 atcctgaact ttatgaaaaa ctgacatctt caaatgaaat aaacaaggag aagagtgaca
     4981 cagttgctga tatagaaagt gaaccagtgg tagaaagcac agaaactgag gggacataaA
     5041 ACCCAGCTTT cttgtacaaa gtggttgata tccagcacag tggcggccgc tcaagtctag
     5101 agggcccgcg gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc
     5161 tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     5221 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     5281 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     5341 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     5401 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     5461 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     5521 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     5581 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5641 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     5701 cacgtaccta gaagttccta ttccgaagtt cctattctct agaaagtata ggaacttcct
     5761 tggccaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg
     5821 acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg
     5881 atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag
     5941 atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca
     6001 ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt
     6061 tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg
     6121 atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag
     6181 gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt
     6241 atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg
     6301 agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg
     6361 gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg
     6421 cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg
     6481 cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc
     6541 cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg
     6601 acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg
     6661 gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg
     6721 gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa
     6781 aggaatagca cgtactacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     6841 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     6901 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     6961 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     7021 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     7081 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     7141 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     7201 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     7261 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     7321 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     7381 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     7441 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     7501 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     7561 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     7621 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     7681 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     7741 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7801 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7861 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7921 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7981 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     8041 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     8101 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     8161 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     8221 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     8281 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     8341 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     8401 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     8461 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     8521 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     8581 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     8641 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     8701 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     8761 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     8821 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     8881 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     8941 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     9001 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     9061 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     9121 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     9181 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     9241 acgtc     
//
