LOCUS       MTK0_036                7588 bp ds-DNA     circular     22-FEB-2019
DEFINITION  .
FEATURES             Location/Qualifiers
     misc_feature    577..577
                     /label="mutation to reduce GC"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     regulatory      3763..4050
                     /label="CAG_enhancer"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer          5360..5379
                     /label="pCAG_F_primer"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     promoter        complement(1705..1852)
                     /label="KanR Promoter"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    612..612
                     /label="mutation to remove ngg"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     promoter        complement(2629..2647)
                     /label="M13_reverse_primer"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     sig_peptide     6571..6600
                     /label="NLS_(zhang)"
                     /ApEinfo_revcolor=#ff0f00
                     /ApEinfo_fwdcolor=#ff0f00
     CDS             5473..6495
                     /label="HPH (Hygromycin Phosphotransferase)"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    586..586
                     /label="mutation to reduce GC"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     misc_feature    209..242
                     /label="FRT"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    485..485
                     /label="mutation to change ngg"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     CDS             complement(889..1704)
                     /label="KanR"
                     /ApEinfo_revcolor=#008000
                     /ApEinfo_fwdcolor=#008000
     misc_feature    704..704
                     /label="mutation to remove ngg"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     CDS             6607..7314
                     /label="mRuby2"
                     /ApEinfo_revcolor=#ff454e
                     /ApEinfo_fwdcolor=#ff454e
     rep_origin      complement(1853..2616)
                     /label="ColE1"
                     /ApEinfo_revcolor=#7f7f7f
                     /ApEinfo_fwdcolor=#7f7f7f
     misc_feature    430..430
                     /label="mutation to remove BsaI cut site"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    247..832
                     /label="Right Arm Homology"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    complement(706..725)
                     /label="gRNA"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_signal     6502..6564
                     /label="T2A"
                     /ApEinfo_revcolor=#0080ff
                     /ApEinfo_fwdcolor=#0080ff
     Terminator      7451..7580
                     /label="SV40-PA"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    2648..3048
                     /label="Left Arm"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    3126..3405
                     /label="HS4"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     terminator      complement(862..888)
                     /label="KanR Terminator"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    543..543
                     /label="mutation to reduce GC content"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     misc_feature    22..204
                     /label="Insulator A1"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    513..513
                     /label="mutation to remove BsmBI cut site"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    3742..5406
                     /label="CAG promotor"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    689..689
                     /label="mutation to reduce GC"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer_bind     833..849
                     /label="M13 Forward (-40) (Elim)"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    7321..7437
                     /label="d4PEST (MODC)"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    3409..3688
                     /label="HS4"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    695..695
                     /label="mutation to reduce GC"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    complement(487..506)
                     /label="gRNA1"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    3053..3086
                     /label="FRT"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    615..615
                     /label="mutation to reduce GC content"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    3704..3737
                     /label="LoxP"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
ORIGIN
        1 gttacagaga tgttacgaac cctgaacagg ttgactatta attgtgtctg cttgatgtgg
       61 acaccaggtg gcgctggaca tcagatttgg agaggcagtt gtctagggaa ccgggctctg
      121 tgccagcgca ggaggcaggc tggctctcct attccaggga tgctcatcca ggaaggaaag
      181 gttgcatgct ggacacacta accttacaGA AGTTCCTATT CtctagaaaG tATAGGAACT
      241 TCGAGTAGAT CgcgggGGAG GGAGGGaTTC TTCtAGGCCC AGGGCGGTCC TTAggAGACa
      301 GGAGGCAGCc GAGAACTCCC ATAAAGGTAT TGCGGCACTC CCCTCCCCCT aCCgAGAAGG
      361 GTGCGGCCTT CTCTCCgCCT CCTCCACtGC AGaTCCCTCA GGATTGCAGC TCGCGCCGaT
      421 TTTcaGAGAT CAcGCaCCTC CCtCCCACAA ACCAGCaGGA CCGACCCCCG CTCCTCCCCC
      481 cACCTCCgCG AGTccgcgag cggatccagc tcATCTCGtC CTTCAGGCGG cGGacgCCCg
      541 GGACGGAGCC gCGGtCGCCG CCAcCCAGAA GcCTCGACCc aGCAGTCCGc ctccgcctcc
      601 gcctccgCCT CTAGTGCGTG CGCAGGGGCG CGGGGcCAGA CACTGCGGCG CggGGCcTCG
      661 GGaAGGACCG TACCAACGaC CGCCTCCCTg CCCCTCCGCc CCCTctgctg acccGCtCAG
      721 TGGTTTCGCT CAattgggac ccgagccaat ggcaaggggc cgttccccgg cttcagcagc
      781 caatcaggtg ccgaagccac acgggcggcg ggtaacccta ctccccgcga gggttttccc
      841 agtcacgacC CGAgcggccg cgtgttacaa ccaattaacc aattctgatt agaaaaactc
      901 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
      961 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
     1021 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
     1081 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
     1141 gaatggcaaa agcttatgca tttctttcca gacttgttca acaggccagc cattacgctc
     1201 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
     1261 gcgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
     1321 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
     1381 ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
     1441 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
     1501 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     1561 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     1621 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tggagcaaga
     1681 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag
     1741 ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga
     1801 cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc agtcatgacc
     1861 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1921 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1981 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2041 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     2101 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2161 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2221 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2281 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2341 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2401 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2461 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2521 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2581 tttcctgcgt tatcccctga ttctgtggat aaccgtgcgg ccgccaatca tggtcatagc
     2641 tgtttccTGC AAcggaaagg gcCTAAAATT TTAAGATTag tatctaattt aaaATGCTAC
     2701 ACTTTGGTGG TGtaaacaac gttctcaaac gacgtgagta aaaaactcag atgaccatgc
     2761 ccttaacctc cttttatcca caggaaaagc actacaaaaa cgatcttaag tagtaataac
     2821 acgcagcctt gcTGCATAAA ATCAGCccga gatgaatgcc tggactcCCA TTTCCCTcCt
     2881 CCACCCcTAT TAtaattatt ttgtgacaca aactcaagca gggtgaagag agaaatctca
     2941 cacCTGCTCA aTCCCCTGCA GGaCAACGCC CAGacaccag gtaaagcctg ggcacagaga
     3001 gTCCCGCCCA cAATCGAGAA GgagCaACTC GACATGGAGG CGATGACGCC CTGAAGTTCC
     3061 TATTCtctag aaaGtATAGG AACTTCctga actggccgat aattgcagac gATCGAATTC
     3121 GAGTTGGCGC GCCTGTCATT CTAAATCTCT CTTTCAGCCT AAAGCTTTTT CCCCGTATCC
     3181 CCCCAGGTGT CTGCAGGCTC AAAGAGCAGC GAGAAGCGTT CAGAGGAAAG CGATCCCGTG
     3241 CCACCTTCCC CGTGCCCGGG CTGTCCCCGC ACGCTGCCGG CTCGGGGATG CGGGGGGAGC
     3301 GCCGGACCGG AGCGGAGCCC CGGGCGGCTC GCTGCTGCCC CCTAGCGGGG GAGGGACGTA
     3361 ATTACATCCC TGGGGGCTTT GGGAGGGGGC TGTCCCCGTG AGCTCCCAGG CGCGCCTGTC
     3421 ATTCTAAATC TCTCTTTCAG CCTAAAGCTT TTTCCCCGTA TCCCCCCAGG TGTCTGCAGG
     3481 CTCAAAGAGC AGCGAGAAGC GTTCAGAGGA AAGCGATCCC GTGCCACCTT CCCCGTGCCC
     3541 GGGCTGTCCC CGCACGCTGC CGGCTCGGGG ATGCGGGGGG AGCGCCGGAC CGGAGCGGAG
     3601 CCCCGGGCGG CTCGCTGCTG CCCCCTAGCG GGGGAGGGAC GTAATTACAT CCCTGGGGGC
     3661 TTTGGGGGGG GGCTGTCCCC GTGAGCTCAA TAAAAGAGCC CACataactt cgtataATGT
     3721 ATGCtatacg aagttataac gCCATATATG GAGTTCCGCG TTACATAACT TACGGTAAAT
     3781 GGCCCGCCTG GCTGACCGCC CAACGACCCC CGCCCATTGA CGTCAATAAT GACGTATGTT
     3841 CCCATAGTAA CGCCAATAGG GACTTTCCAT TGACGTCAAT GGGTGGAGTA TTTACGGTAA
     3901 ACTGCCCACT TGGCAGTACA TCAAGTGTAT CATATGCCAA GTACGCCCCC TATTGACGTC
     3961 AATGACGGTA AATGGCCCGC CTGGCATTAT GCCCAGTACA TGACCTTATG GGACTTTCCT
     4021 ACTTGGCAGT ACATCTACGT ATTAGTCATC GCTATTACCA TGGTCGAGGT GAGCCCCACG
     4081 TTCTGCTTCA CTCTCCCCAT CTCCCCCCCC TCCCCACCCC CAATTTTGTA TTTATTTATT
     4141 TTTTAATTAT TTTGTGCAGC GATGGGGGCG GGGGGGGGGG GGGCGCGCGC CAGGCGGGGC
     4201 GGGGCGGGGC GAGGGGCGGG GCGGGGCGAG GCGGAGAGGT GCGGCGGCAG CCAATCAGAG
     4261 CGGCGCGCTC CGAAAGTTTC CTTTTATGGC GAGGCGGCGG CGGCGGCGGC CCTATAAAAA
     4321 GCGAAGCGCG CGGCGGGCGG GAGTCGCTGC GTTGCCTTCG CCCCGTGCCC CGCTCCGCGC
     4381 CGCCTCGCGC CGCCCGCCCC GGCTCTGACT GACCGCGTTA CTCCCACAGG TGAGCGGGCG
     4441 GGACGGCCCT TCTCCTCCGG GCTGTAATTA GCGCTTGGTT TAATGACGGC TCGTTTCTTT
     4501 TCTGTGGCTG CGTGAAAGCC TTAAAGGGCT CCGGGAGGGC CCTTTGTGCG GGGGGGAGCG
     4561 GCTCGGGGGG TGCGTGCGTG TGTGTGTGCG TGGGGAGCGC CGCGTGCGGC CCGCGCTGCC
     4621 CGGCGGCTGT GAGCGCTGCG GGCGCGGCGC GGGGCTTTGT GCGCTCCGCG TGTGCGCGAG
     4681 GGGAGCGCGG CCGGGGGCGG TGCCCCGCGG TGCGGGGGGG CTGCGAGGGG AACAAAGGCT
     4741 GCGTGCGGGG TGTGTGCGTG GGGGGGTGAG CAGGGGGTGT GGGCGCGGCG GTCGGGCTGT
     4801 AACCCCCCCC TGCACCCCCC TCCCCGAGTT GCTGAGCACG GCCCGGCTTC GGGTGCGGGG
     4861 CTCCGTaCGG GGCGTGGCGC GGGGCTCGCC GTGCCGGGCG GGGGGTGGCG GCAGGTGGGG
     4921 GTGCCGGGCG GGGCGGGGCC GCCTCGGGCC GGGGAGGGCT CGGGGGAGGG GCGCGGCGGc
     4981 CCCCGGAGtG CCGGCGGCTG TCGAGGCGCG GCGAGCCGCA GCCATTGCCT TTTATGGTAA
     5041 TCGTGCGAGA GGGCGCAGGG ACTTCCTTTG TCCCAAATCT GTGCGGAGCC GAAATCTGGG
     5101 AGGCGCCGCC GCACCCCCTC TAGCGGGCGC gGGGCGAAGC GGTGCGGCGC CGGCAGGAAG
     5161 GAAATGGGCG GGGAGGGCCT TCGTGCGTCG CCGCGCCGCC GTCCCCTTCT CCcTCTCCAG
     5221 CCTCGGGGCT GTCCGCgGGG GGACGGCTGC CTTCGGGGGG GACGGGGCAG GGCGGGGTTC
     5281 GGCTTCTGGC GTGTGACCGG CGGCTCTAGA GCCTCTGCTA ACCATGTTCA TGCCTTCTTC
     5341 TTTTTCCTAC AGCTCCTGGG CAACGTGCTG GTTATTGTGC TGTCTCATCA TTTTGGCAAA
     5401 Gaattcgcca cTATGGTGGT TTGTCTGGTC AACCACCgcg gtctcAGTGG TGTACGGTAC
     5461 AAACCAGGTT CTggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc
     5521 gaaaagttcg acagcgtgtc cgacctgatg cagctctcgg agggcgaaga atctcgtgct
     5581 ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt
     5641 ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa
     5701 gtgcttgaca ttggggaatt tagcgagagc ctgacctatt gcatctcccg ccgtgcacag
     5761 ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca accggtcgcg
     5821 gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc
     5881 ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat
     5941 ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag
     6001 gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac
     6061 gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac
     6121 tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg
     6181 ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt
     6241 gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag
     6301 agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc
     6361 gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc
     6421 tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt
     6481 ccgagggcaa aggaaGGATC CGGAAGCGGA GAGGGCAGAG GAAGTCTGCT AACATGCGGT
     6541 GACGTCGAGG AGAATCCTGG ACCTGGATCC CCCAAGAAGA AGAGGAAGGT GGAGGACGCC
     6601 GGATCCgtgt ccaaaggaga ggagttaatc aaggaaaaca tgagaatgaa agttgtcatg
     6661 gagggctccg ttaatggtca ccaattcaag tgtacagggg aaggtgaagg taatccttac
     6721 atgggtacac aaactatgag aattaaagta attgaaggcg gaccactacc atttgcattt
     6781 gacattctgg caacgtcatt catgtacgga tcacgaactt tcatcaagta ccctaaaggt
     6841 ataccagact ttttcaagca atcttttcca gagggtttta catgggaaag ggttacaaga
     6901 tacgaagatg ggggtgtcgt cacagttatg caagatactt cattagaaga tggctgcctt
     6961 gtctatcatg tgcaagtaag aggggtgaat tttccttcta acggacctgt gatgcagaaa
     7021 aagaccaaag gttgggaacc aaatactgaa atgatgtacc cagctgatgg aggtttgaga
     7081 ggctacacac acatggcgct taaagttgat ggtggaggtc atttgtcttg tagttttgtt
     7141 accacttatc gttctaaaaa gactgttggc aatatcaaaa tgccaggaat acatgctgta
     7201 gaccacagac tagaaagact cgaagagagc gataacgaaa tgttcgttgt acagagagag
     7261 catgccgtag ccaaatttgc tggcttaggc ggtggtatgg atgaattgta taagggaTCC
     7321 CATGGCTTCC CGCCGGAGGT GGAGGAGCAG GATGATGGCG CGCTGCCCAT GTCTTGTGCC
     7381 CAGGAGAGCG GGATGGACCG TCACCCTGCA GCCTGTGCTT CTGCTAGGAT CAATGTGTAA
     7441 CTCGAGTGGC gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     7501 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     7561 cattataagc tgcaataaac GCTGagca
//