LOCUS       824-U6-spTet-CMV        7644 bp ds-DNA     circular     23-SEP-2017
DEFINITION  .
ACCESSION   urn.local...83by053-2n
VERSION
KEYWORDS    .
SOURCE      
  ORGANISM  .
            .
COMMENT     U6_sgRNA_template from 1 to 8308 pMig-w from 1 to 6471 
            ApEinfo:methylated:1.
FEATURES             Location/Qualifiers
     misc_feature    263..826
                     /ApEinfo_fwdcolor="#fffc15"
                     /ApEinfo_revcolor="#fffc15"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="pU6 (pMJ177) 5' CMV promoter IE"
     misc_feature    complement(766..775)
                     /ApEinfo_fwdcolor="#888eff"
                     /ApEinfo_revcolor="#888eff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="GC Box"
     misc_feature    844..1024
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="cyan"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="HIV-1_5_LTR"
     misc_feature    complement(1601..1607)
                     /ApEinfo_fwdcolor="#ff9afa"
                     /ApEinfo_revcolor="#ff9afa"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="TATA box"
     misc_feature    1695..1928
                     /ApEinfo_fwdcolor="#9cffe3"
                     /ApEinfo_revcolor="#9cffe3"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="RRE"
     misc_feature    2459..2473
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="cyan"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="cPPT"
     misc_feature    2631..2944
                     /ApEinfo_label="mU6 promoter (modified)"
                     /ApEinfo_fwdcolor="#fff431"
                     /ApEinfo_revcolor="#fff431"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="mU6 promoter (modified)(1)"
     misc_recomb     2825..2858
                     /ApEinfo_fwdcolor="steel blue"
                     /ApEinfo_revcolor="orange"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="LoxP recombination site"
     misc_feature    join(2919,2934..2936,2940..2944)
                     /ApEinfo_fwdcolor="#fff431"
                     /ApEinfo_revcolor="#fff431"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="mU6 promoter (modified)"
     sgRNA           2945..2965
                     /SerialCloner_Color="&hDC143C"
                     /SerialCloner_Show="True"
                     /SerialCloner_Protect="True"
                     /SerialCloner_Arrow="True"
                     /SerialCloner_Desc="ApEinfo_graphicformat=arrow_data {{0 1
                     2 0 0 -1} {} 0} - width 5 offset 0 - label=sgTet3g -
                     ApEinfo_fwdcolor=#800080 - ApEinfo_revcolor=green -"
                     /ApEinfo_fwdcolor="#cd3249"
                     /ApEinfo_revcolor="#eb1439"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /modified_by="User"
                     /label="sgTRE3G"
     misc_feature    2966..3058
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="modified stem loop"
     misc_feature    3116..3698
                     /ApEinfo_fwdcolor="#ffed31"
                     /ApEinfo_revcolor="#ffed31"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="CMV promoter IE"
     misc_feature    3116..3679
                     /ApEinfo_label="pU6 (pMJ177) 5' CMV promoter IE"
                     /ApEinfo_fwdcolor="#fffc15"
                     /ApEinfo_revcolor="#fffc15"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="pU6 (pMJ177) 5' CMV promoter IE(1)"
     misc_feature    complement(3619..3628)
                     /ApEinfo_label="GC Box"
                     /ApEinfo_fwdcolor="#888eff"
                     /ApEinfo_revcolor="#888eff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="GC Box(1)"
     GFP             3717..>4421
                     /created_by="User"
                     /label="mTagBFP2"
     source          <3717..>4421
                     /organism="pEGIP"
                     /mol_type="other DNA"
                     /label="source pEGIP"
     misc_feature    3766..3771
                     /Recognition_pattern="T^GTACA"
                     /annotation_group="TGTACA"
                     /label="BsrGI"
                     /note="Geneious type: restriction site"
     misc_feature    <4422..4424
                     /ApEinfo_label="GFP"
                     /ApEinfo_fwdcolor="#00ff00"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="GFP(2)"
     misc_recomb     4444..4477
                     /ApEinfo_label="LoxP recombination site"
                     /ApEinfo_fwdcolor="steel blue"
                     /ApEinfo_revcolor="orange"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="LoxP recombination site(1)"
     misc_feature    4523..5123
                     /ApEinfo_fwdcolor="#0060ff"
                     /ApEinfo_revcolor="#0060ff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="wPRE"
     misc_feature    5313..5327
                     /ApEinfo_label="cPPT"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="cyan"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="cPPT(1)"
     misc_feature    5650..5830
                     /ApEinfo_label="HIV-1_5_LTR"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="cyan"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="HIV-1_5_LTR(1)"
     rep_origin      complement(5874..6556)
                     /ApEinfo_fwdcolor="gray50"
                     /ApEinfo_revcolor="gray50"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="ColE1 origin"
     misc_feature    complement(5877..6496)
                     /ApEinfo_fwdcolor="#95928e"
                     /ApEinfo_revcolor="#95928e"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="pBR322_origin"
     misc_feature    complement(5901..5910)
                     /ApEinfo_label="GC Box"
                     /ApEinfo_fwdcolor="#888eff"
                     /ApEinfo_revcolor="#888eff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="GC Box(2)"
     CDS             complement(6654..7313)
                     /ApEinfo_fwdcolor="#ffff00"
                     /ApEinfo_revcolor="#ffff00"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="AmpR"
     misc_feature    complement(7553..7581)
                     /ApEinfo_fwdcolor="#ffff00"
                     /ApEinfo_revcolor="#ffff00"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="AmpR promoter"
ORIGIN      
        1 gcttaagcgg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
       61 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
      121 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
      181 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      241 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      301 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      361 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      421 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      481 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      541 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      601 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      661 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      721 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      781 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
      841 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
      901 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
      961 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     1021 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     1081 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     1141 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     1201 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     1261 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     1321 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     1381 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     1441 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     1501 agcaaaacaa aagtaagacc accgcacagc aagcggccgg ccgcgctgat cttcagacct
     1561 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
     1621 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
     1681 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
     1741 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
     1801 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
     1861 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
     1921 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
     1981 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
     2041 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
     2101 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
     2161 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
     2221 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
     2281 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc
     2341 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac
     2401 ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa
     2461 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
     2521 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta
     2581 ttacagggac agcagagatc cagtttggtt agtaccgggc ccgctctaga gatccgacgc
     2641 gccatctcta ggcccgcgcc ggccccctcg cacggacttg tgggagaagc tcggctactc
     2701 ccctgccccg gttaatttgc atataatatt tcctagtaac tatagaggct taatgtgcga
     2761 taaaagacag ataatctgtt ctttttaata ctagctacat tttacatgat aggcttggat
     2821 ttctataact tcgtatagca tacattatac gaagttataa acagcacaaa aggaaactca
     2881 ccctaactgt aaagtaattg tgtgttttga gactataagt atcccttgga gaaccacctt
     2941 gttggtacgt tctctatcac tgatagttta agagctatgc tggaaacagc atagcaagtt
     3001 taaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttttct
     3061 cgagtactag gatccattag gcggccgcgt ggataaccgt attaccgcca tgcattagtt
     3121 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     3181 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     3241 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     3301 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     3361 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     3421 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     3481 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     3541 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     3601 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     3661 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag cccaccatgg
     3721 tgtctaaggg cgaagagctg attaaggaga acatgcacat gaagctgtac atggagggca
     3781 ccgtggacaa ccatcacttc aagtgcacat ccgagggcga aggcaagccc tacgagggca
     3841 cccagaccat gagaatcaag gtggtcgagg gcggccctct ccccttcgcc ttcgacatcc
     3901 tggctactag cttcctctac ggcagcaaga ccttcatcaa ccacacccag ggcatccccg
     3961 acttcttcaa gcagtccttc cctgagggct tcacatggga gagagtcacc acatacgaag
     4021 acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggctgc ctcatctaca
     4081 acgtcaagat cagaggggtg aacttcacat ccaacggccc tgtgatgcag aagaaaacac
     4141 tcggctggga ggccttcacc gagacgctgt accccgctga cggcggcctg gaaggcagaa
     4201 acgacatggc cctgaagctc gtgggcggga gccatctgat cgcaaacgcc aagaccacat
     4261 atagatccaa gaaacccgct aagaacctca agatgcctgg cgtctactat gtggactaca
     4321 gactggaaag aatcaaggag gccaacaacg agacctacgt cgagcagcac gaggtggcag
     4381 tggccagata ctgcgacctc cctagcaaac tggggcacaa ataagaattc gtcgagggac
     4441 ctaataactt cgtatagcat acattatacg aagttataca tgtttaaggg ttccggttcc
     4501 actaggtaca attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
     4561 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     4621 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     4681 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     4741 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     4801 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     4861 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     4921 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     4981 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     5041 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     5101 ccctttgggc cgcctccccg catcgatacc gtcgacctcg atcgagacct agaaaaacat
     5161 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca
     5221 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact
     5281 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     5341 attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac
     5401 ttccctgatt ggcagaacta cacaccaggg ccagggatca gatatccact gacctttgga
     5461 tggtgctaca agctagtacc agttgagcaa gagaaggtag aagaagccaa tgaaggagag
     5521 aacacccgct tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagta
     5581 ttagagtgga ggtttgacag ccgcctagca tttcatcaca tggcccgaga gctgcatccg
     5641 gactgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     5701 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     5761 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     5821 atctctagca gcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5881 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5941 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6001 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6061 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6121 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6181 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6241 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6301 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6361 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6421 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6481 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6541 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6601 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6661 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6721 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6781 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6841 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6901 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6961 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7021 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7081 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7141 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7201 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7261 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7321 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7381 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7441 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7501 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7561 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7621 tttccccgaa aagtgccacc tgac
//
