LOCUS       Exported                8925 bp ds-DNA     circular SYN 19-JAN-2016
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8925)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jul 19, 2019 from SnapGene 4.3.10
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8925
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    263..826
                     /label=pU6 (pMJ177) 5' CMV promoter IE
                     /note="color: #fffc15"
     misc_feature    complement(766..775)
                     /label=GC Box
                     /note="color: #888eff; direction: LEFT"
     misc_feature    844..1024
                     /label=HIV-1_5_LTR
                     /note="color: #00ffff"
     misc_feature    complement(1601..1607)
                     /label=TATA box
                     /note="color: #ff9afa; direction: LEFT"
     misc_feature    1695..1928
                     /label=RRE
                     /note="color: #9cffe3"
     misc_feature    2459..2473
                     /label=cPPT
                     /note="color: #00ffff"
     misc_feature    2639..2952
                     /label=mU6 promoter (modified)
                     /label=mU6 promoter (modified)(1)
                     /note="color: #fff431"
     misc_recomb     2833..2866
                     /label=LoxP recombination site
                     /note="color: #4682b4"
     misc_feature    2872..2892
                     /label=Seq_5'
                     /note="color: #00ffff"
     misc_feature    2908..2953
                     /label=Sequencing primer
                     /note="color: #7f007f"
     misc_feature    join(2927,2942..2944,2948..2952)
                     /label=mU6 promoter (modified)
                     /note="This feature has 3 segments:
                        1: 2927 .. 2927 / #fff431
                        2: 2942 .. 2944 / #fff431
                        3: 2948 .. 2952 / #fff431"
     misc_feature    2953..2972
                     /label=EGFP_NT2
                     /note="color: #ffa1e9"
     misc_feature    2973..2984
                     /label=modified stem loop
                     /note="color: #00ffff"
     misc_feature    2986..2997
                     /label=modified stem loop
                     /label=modified stem loop(1)
                     /note="color: #00ffff"
     misc_feature    2999..3065
                     /label=modified stem loop
                     /label=modified stem loop(2)
                     /note="color: #00ffff"
     misc_feature    3035..3054
                     /label=Seq_3'
                     /note="color: #00ffff"
     misc_feature    3035..3054
                     /label=Seq_3'
                     /label=Seq_3'(1)
                     /note="color: #ff7f00"
     misc_feature    3131..4301
                     /label=hEF1a promoter
                     /note="color: #2c9c57"
     misc_feature    4093
                     /label=SNP found in both clones
                     /note="color: #ff0000"
     misc_feature    4317..4973
                     /label=puro-t2A
                     /note="color: #ca72ff"
     misc_feature    4533..4539
                     /label=RR47
                     /note="color: #ffa1e9"
     misc_feature    4534..4539
                     /label=RR48
                     /note="color: #ffa1e9"
     misc_feature    4911..4985
                     /label=T2A
                     /note="color: #00ffff"
     misc_feature    4986..5702
                     /label=DeWitt BFP ORF
                     /note="color: #0000ff"
     misc_feature    5150
                     /label=C->g silent mutation
                     /note="color: #ff0000"
     misc_feature    5183..5186
                     /label=cCAT (BFP) -> gTAC (GFP)
                     /note="color: #ff0000"
     misc_feature    complement(5185..5204)
                     /label=Corrected L2 Guide
                     /note="color: #00ff00; direction: LEFT"
     misc_feature    5187..5188
                     /label=Approximate location of cut
                     /note="color: #00ffff"
     misc_feature    5612
                     /label=Silent mutation to disrupt BlpI site
                     /note="color: #ff0000"
     misc_recomb     5725..5758
                     /label=LoxP recombination site
                     /label=LoxP recombination site(1)
                     /note="color: #4682b4"
     misc_feature    5804..6404
                     /label=wPRE
                     /note="color: #0060ff"
     misc_feature    6594..6608
                     /label=cPPT
                     /label=cPPT(1)
                     /note="color: #00ffff"
     misc_feature    6931..7111
                     /label=HIV-1_5_LTR
                     /label=HIV-1_5_LTR(1)
                     /note="color: #00ffff"
     rep_origin      complement(7155..7837)
                     /direction=LEFT
                     /label=ColE1 origin
                     /note="color: #808080"
     misc_feature    complement(7158..7777)
                     /label=pBR322_origin
                     /note="color: #95928e; direction: LEFT"
     misc_feature    complement(7182..7191)
                     /label=GC Box
                     /label=GC Box(1)
                     /note="color: #888eff; direction: LEFT"
     CDS             complement(7935..8594)
                     /label=AmpR
                     /note="color: #ffff00; direction: LEFT"
     misc_feature    complement(8834..8862)
                     /label=AmpR promoter
                     /note="color: #ffff00; direction: LEFT"
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 ccgccctgca ggtctagaga
     2641 tccgacgcgc catctctagg cccgcgccgg ccccctcgca cggacttgtg ggagaagctc
     2701 ggctactccc ctgccccggt taatttgcat ataatatttc ctagtaacta tagaggctta
     2761 atgtgcgata aaagacagat aatctgttct ttttaatact agctacattt tacatgatag
     2821 gcttggattt ctataacttc gtatagcata cattatacga agttataaac agcacaaaag
     2881 gaaactcacc ctaactgtaa agtaattgtg tgttttgaga ctataagtat cccttggaga
     2941 accaccttgt tggaccagga tgggcaccac ccgtttaaga gctaagctgg aaacagcata
     3001 gcaagtttaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
     3061 tttttctcga gtactaggat ccattaggcg gccgccctgc agggtggata accgtattac
     3121 cgccatgcat gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
     3181 gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa
     3241 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
     3301 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt
     3361 gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga
     3421 attacttcca cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg
     3481 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc
     3541 tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct
     3601 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc
     3661 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg
     3721 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg
     3781 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg
     3841 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca
     3901 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg
     3961 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt
     4021 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc
     4081 gattagttct cgcgcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg
     4141 atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg
     4201 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag
     4261 acagtggttc aaagtttttt tcttccattt caggtgtcgt gagctagccc accatgaccg
     4321 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc
     4381 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca
     4441 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     4501 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     4561 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     4621 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     4681 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     4741 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     4801 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     4861 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc ggatcgggag
     4921 agggcagagg aagtctgcta acatgcggtg acgtcgagga gaatcctggc ccaccggtcg
     4981 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     5041 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     5101 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccg gtgccctggc
     5161 ccaccctcgt gaccaccctg acccatggcg tgcagtgctt cagccgctac cccgaccaca
     5221 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     5281 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     5341 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     5401 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     5461 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     5521 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     5581 accactacct gagcacccag tccaagctga gtaaagaccc caacgagaag cgcgatcaca
     5641 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     5701 agtaagaatt cgtcgaggga cctaataact tcgtatagca tacattatac gaagttatac
     5761 atgtttaagg gttccggttc cactaggtac aattcgatat caagcttatc gataatcaac
     5821 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     5881 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     5941 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     6001 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     6061 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     6121 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     6181 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     6241 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     6301 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     6361 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc
     6421 gatcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct
     6481 gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag
     6541 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     6601 aaggggggac tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg
     6661 atctaccaca cacaaggcta cttccctgat tggcagaact acacaccagg gccagggatc
     6721 agatatccac tgacctttgg atggtgctac aagctagtac cagttgagca agagaaggta
     6781 gaagaagcca atgaaggaga gaacacccgc ttgttacacc ctgtgagcct gcatgggatg
     6841 gatgacccgg agagagaagt attagagtgg aggtttgaca gccgcctagc atttcatcac
     6901 atggcccgag agctgcatcc ggactgtact gggtctctct ggttagacca gatctgagcc
     6961 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     7021 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     7081 cccttttagt cagtgtggaa aatctctagc agcatgtgag caaaaggcca gcaaaaggcc
     7141 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     7201 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7261 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7321 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     7381 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7441 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7501 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7561 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     7621 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7681 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7741 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7801 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7861 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7921 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7981 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     8041 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     8101 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     8161 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     8221 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     8281 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     8341 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     8401 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     8461 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8521 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8581 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8641 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8701 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8761 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8821 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8881 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgac
//
