LOCUS       Exported                8738 bp ds-DNA     circular UNA 10-FEB-2017
DEFINITION  natural circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 17
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8738)
  AUTHORS   Addgene
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Feb 10, 2017 from SnapGene 3.3.2
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8738
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     misc_feature    1..515
                     /label=5'LTR
                     /note="5'LTR"
     misc_feature    516..1406
                     /label=Psi
                     /note="Psi"
     promoter        1445..1693
                     /label=hU6
                     /label=hU6 promoter
                     /note="hU6 promoter"
     misc_feature    1696..1699
                     /label=BbsI overhang
                     /note="BbsI overhang"
     Site            order(1702..1707,3160..3165)
                     /note="BbsI"
     promoter        1732..1755
                     /label=Lac Pro
                     /label=Lac Promoter
                     /note="Lac Pro promoter"
     CDS             1816..2475
                     /label=CmR
                     /label=Chloramphenicol resistance
                     /note="CmR"
     CDS             2817..3122
                     /label=ccdB
                     /label=ccdB gene
                     /note="ccdB"
     misc_feature    3168..3249
                     /label=Guide RNA
                     /label=sgRNA scaffold
                     /note="Guide RNA"
     misc_feature    complement(3168..3171)
                     /label=BbsI overhang
                     /note="BbsI overhang"
     promoter        3362..3869
                     /label=PGK
                     /label=PGK promoter
                     /note="PGK promoter"
     misc_feature    3883..3888
                     /label=NheI
                     /note="NheI"
     CDS             3894..4490
                     /label=PuroR
                     /label=Puromycin resistance
                     /note="PuroR"
     misc_feature    4491..4544
                     /label=T2A
                     /label=T2A peptide
                     /note="T2A"
     CDS             4551..5270
                     /label=BFP
                     /note="BFP"
     misc_feature    5271..5276
                     /label=MluI(1)
                     /note="MluI(1)"
     misc_feature    5322..5644
                     /label=3'LTR
                     /note="3'LTR"
     CDS             6944..7804
                     /label=AmpR
                     /label=Ampicillin resistance (bla)
                     /note="AmpR"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gatcgatctc tcgacctaga gatatcgtgt
     1441 cgacccccac gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc
     1501 tgttagagag ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac
     1561 gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat
     1621 ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt
     1681 gtggaaagga cgaaacaccg ggtcttcgcg gccgcattag gcaccccagg ctttacactt
     1741 tatgcttccg gctcgtataa tgtgtggatt ttgagttagg atccgtcgag attttcagga
     1801 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa
     1861 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
     1921 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
     1981 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg
     2041 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc
     2101 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag
     2161 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct
     2221 aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt
     2281 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa
     2341 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt
     2401 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg
     2461 cagggcgggg cgtaaacgcg tggatccggc ttactaaaag ccagataaca gtatgcgtat
     2521 ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca
     2581 aaaagaggta tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt
     2641 tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga
     2701 agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt
     2761 cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct ggtgaaatgc
     2821 agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga
     2881 gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc
     2941 tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc
     3001 gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg
     3061 atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat
     3121 aaatgtcagg ctcccttata cacagccagt ctgcaggtcg aagacctgtt ttagagctag
     3181 aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg
     3241 tgcttttttg ttttagagct agaaatagca agttaaaata aggctagtcc gtttttagcg
     3301 cgtgcgccaa ttctgcagac aaatggctcc tcgagaattc gcgaatcgat gttaacgcgt
     3361 aattctaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag
     3421 ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca
     3481 ccggtaggcg gcaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc
     3541 ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg
     3601 aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg
     3661 taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc
     3721 tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc
     3781 gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt
     3841 tctcctcttc ctcatctccg ggcctttcga cctgcagccc aagctagctt accatgaccg
     3901 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     3961 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     4021 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     4081 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     4141 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     4201 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     4261 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     4321 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     4381 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     4441 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc gagggcagag
     4501 gaagtctgct aacatgcggt gacgtggagg agaatcccgg ccctgctagc atggtgagca
     4561 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     4621 acggccacaa gttcagcgtg aggggcgagg gcgagggcga tgccaccaac ggcaagctga
     4681 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     4741 ccctgagcca cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     4801 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     4861 acggcaccta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     4921 tcgagctgaa gggcgtcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     4981 acaacttcaa cagccacaac atctatatca tggccgtcaa gcagaagaac ggcatcaagg
     5041 tgaacttcaa gatccgccac aacgtggagg acggcagcgt gcagctcgcc gaccactacc
     5101 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacagccac tacctgagca
     5161 cccagtccgt gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     5221 tccgcaccgc cgccgggatc actctcggca tggacgagct gtacaagtga acgcgtcgat
     5281 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
     5341 gtttggcaag ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
     5401 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
     5461 tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgataga
     5521 ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
     5581 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
     5641 ttcatgggta acagtttctt gaagttggag aacaacattc tgagggtagg agtcgaatat
     5701 taagtaatcc tgactcaatt agccactgtt ttgaatccac atactccaat actcctgaaa
     5761 tagttcatta tggacagcgc agaaagagct ggggagaatt gtgaaattgt tatcccctca
     5821 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5881 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5941 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6001 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6061 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6121 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6181 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6241 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6301 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6361 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6421 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6481 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6541 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6601 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6661 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6721 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6781 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6841 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6901 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6961 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7021 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7081 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7141 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7201 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7261 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7321 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7381 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7441 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7501 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7561 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7621 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7681 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7741 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7801 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7861 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7921 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     7981 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     8041 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     8101 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     8161 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     8221 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     8281 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     8341 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggcgcaag
     8401 gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat
     8461 acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt
     8521 gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat
     8581 gcgtccggcg tagaggcgat tagtccaatt tgttaaagac aggatatcag tggtccaggc
     8641 tctagttttg actcaacaat atcaccagct gaagcctata gagtacgagc catagataaa
     8701 ataaaagatt ttatttagtc tccagaaaaa ggggggaa
//
