LOCUS       APP8_beta_arrest        5773 bp ds-DNA     circular     09-NOV-2017
SOURCE      
  ORGANISM  
COMMENT     NewMCS:XbaI,BamHI,NheI,AgeI,BstBI,AfeI(blunt),HpaI(blunt),AscI(8cut
            ter),SbfI,EcoRIOtherpossibleuniquecutters(currentlynosites)BsiWI,Bm
            tI(samesequenceasNheI)Tochangepromoter cutwith PacIand XbaI
COMMENT     
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     misc_feature    3037..3177
                     /label=ITR
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    753..769
                     /label=T7 promoter primer
                     /ApEinfo_fwdcolor="#00ff00"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    785..786
                     /label=NotI
                     /ApEinfo_fwdcolor="#ff8040"
                     /ApEinfo_revcolor="#ff8040"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2777..3008
                     /label=PolyA
                     /ApEinfo_fwdcolor="#ff2549"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    151..708
                     /label=CMV promoter
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2741..2758
                     /label=S219V-tev in BL21 DNA
                     /ApEinfo_fwdcolor="#a486ff"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    791..793
                     /label=Kozak consensus 
                     /ApEinfo_fwdcolor="#ffd92b"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    791..793
                     /label=start codon
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    join(2288..2739,2741..2758)
                     /label=TEV for yeast
                     /ApEinfo_fwdcolor="#ff51d8"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1..130
                     /label=ITR(1)
                     /ApEinfo_label="ITR"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2105..2287
                     /label=S219V-tev in BL21 DNA(1)
                     /ApEinfo_label="S219V-tev in BL21 DNA"
                     /ApEinfo_fwdcolor="#a486ff"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2765..2770
                     /label=EcoRI
                     /ApEinfo_fwdcolor="#46ff6e"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    794..2020
                     /label=rat beta arrestin 2
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2288..2739
                     /label=S219V-tev in BL21 DNA(2)
                     /ApEinfo_label="S219V-tev in BL21 DNA"
                     /ApEinfo_fwdcolor="#a486ff"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    join(2105..2248,2255..2287)
                     /label=TEV for yeast(1)
                     /ApEinfo_label="TEV for yeast"
                     /ApEinfo_fwdcolor="#ff51d8"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2075..2104
                     /label=GGSGSGGSGSGSGGS linker
                     /ApEinfo_fwdcolor="#fff679"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2021..2047
                     /label=HA tag
                     /ApEinfo_fwdcolor="#3a23ff"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtTGTACA tagtaatcaa ttacggggtc attagttcat
      181 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      241 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      301 gggactttcc attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta
      361 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      421 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      481 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      541 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      601 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      661 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact
      721 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa
      781 gcttgccacc atgGGTGAAA AACCCGGGAC CAGGGTCTTC AAGAAGTCGA GCCCTAACTG
      841 CAAGCTCACC GTGTACTTGG GCAAGCGTGA CTTTGTGGAT CACTTGGACA AAGTGGATCC
      901 TGTCGATGGT GTGGTGCTTG TGGATCCTGA CTACTTGAAG GACCGGAAAG TGTTTGTGAC
      961 CCTCACCTGT GCCTTCCGCT ATGGCCGAGA AGACCTGGAT GTACTGGGCC TGTCTTTCCG
     1021 CAAAGATCTG TTCATCGCCA CCTACCAGGC CTTCCCCCCC ATGCCCAACC CACCTCGGCC
     1081 CCCCACCCGC CTACAGGACC GACTGCTGAA GAAGTTGGGC CAGCAcGCCC ACCCCTTTTT
     1141 TTTCACAATA CCCCAGAATT TGCCTTGCTC CGTCACACTG CAGCCAGGAC CGGAGGACAC
     1201 AGGGAAGGCC TGTGGAGTAG ACTTTGAGAT TCGAGCCTTC TGTGCCAAAT CTATAGAAGA
     1261 AAAAAGCCAC AAAAGGAACT CCGTGCGGCT TATCATCAGA AAGGTACAGT TTGCTCCTGA
     1321 GACACCCGGC CCCCAGCCAT CAGCTGAAAC CACACGCCAC TTCCTCATGT CTGACCGGAG
     1381 GTCCCTGCAC CTAGAGGCTT CCCTGGACAA AGAGCTGTAC TACCATGGGG AACCCCTCAA
     1441 TGTCAACGTC CACGTCACCA ACAATTCTGC CAAaACCGTC AAGAAGATCA GAGTGTCTGT
     1501 GAGACAGTAT GCCGACATTT GCCTCTTCAG CACCGCGCAG TACAAGTGTC CTGTGGCTCA
     1561 GCTTGAACAA GATGACCAGG TGTCTCCCAG TTCCACATTC TGCAAGGTGT ACACCATAAC
     1621 CCCGCTGCTC AGTGACAACC GAGAGAAGCG TGGCCTTGCC CTTGATGGGC AACTCAAGCA
     1681 CGAAGACACC AACCTGGCTT CCAGCACCAT TGTGAAGGAG GGAGCCAACA AGGAGGTGCT
     1741 GGGAATCCTA GTATCCTACA GGGTCAAGGT GAAGCTGGTG GTGTCTCGAG GCGGGGATGT
     1801 CTCCGTGGAG CTACCTTTCG TCCTAATGCA CCCCAAGCCC CACGACCACA TCACCCTTCC
     1861 CCGACCCCAG TCAGCCCCCC GGGAAATAGA CATCCCTGTG GATACCAACC TCATTGAATT
     1921 CGATACCAAC TATGCCACAG ACGACGACAT CGTGTTTGAG GACTTTGCGA GGCTTCGGCT
     1981 GAAGGGGATG AAGGATGACG ACTGTGATGA CCAGTTCTGC TACCCATACG ATGTGCCAGA
     2041 TTACGCCtat ccctatgacg tccctgatta cgctGGTGGA AGTGGATCAG GCAGCGGTGG
     2101 ATCTGGAGAA AGCttgttta agggaccacg tgattacaac ccgatatcga gcaccatttg
     2161 tcatttgacg aatgaatctg atgggcacac aacatcgttg tatggtattg gatttggtcc
     2221 cttcatcatt acaaacaagc acttgtttAG AAGAaataat ggaacactgt tggtccaatc
     2281 actacatggt gtattcaagg tcaagaacac cacgactttg caacaacacc tcattgatgg
     2341 gagggacatg ataattattc gcatgcctaa ggatttccca ccatttcctc aaaagctgaa
     2401 atttagagag ccacaaaggg aagagcgcat atgtcttgtg acaaccaact tccaaactaa
     2461 gagcatgtct agcatggtgt cagacactag ttgcacattc ccttcatctg atggcatatt
     2521 ctggaagcat tggattcaaa ccaaggatgg gcagtgtggc agtccattag tatcaactag
     2581 agatgggttc attgttggta tacactcagc atcgaatttc accaacacaa acaattattt
     2641 cacaagcgtg ccgaaaaact tcatggaatt gttgacaaat caggaggcgc agcagtgggt
     2701 tagtggttgg cgattaaatg ctgactcagt attgtggggt ggccataaag ttttcatggt
     2761 gTAAGAATTC ctcgagCTAG AGCTCGCTGA TCAGCCTCGA CTGTGCCTTC TAGTTGCCAG
     2821 CCATCTGTTG TTTGCCCCTC CCCCGTGCCT TCCTTGACCC TGGAAGGTGC CACTCCCACT
     2881 GTCCTTTCCT AATAAAATGA GGAAATTGCA TCGCATTGTC TGAGTAGGTG TCATTCTATT
     2941 CTGGGGGGTG GGGTGGGGCA GGACAGCAAG GGGGAGGATT GGGAAGACAA TAGCAGGCAT
     3001 GCTGGGGAgg taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     3061 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     3121 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     3181 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat tcacaccgca tacgtcaaag
     3241 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3301 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3361 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3421 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     3481 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3541 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc gggctattct
     3601 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3661 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     3721 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     3781 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     3841 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     3901 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     3961 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4021 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4081 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4141 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4201 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4261 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4321 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4381 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4441 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4501 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     4561 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4621 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4681 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4741 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4801 gagcgtggaa gccgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     4861 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4921 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4981 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5041 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5101 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5161 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5221 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     5281 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5341 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5401 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5461 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5521 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5581 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5641 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5701 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5761 tttgctcaca tgt    
//
