LOCUS       Exported                8789 bp ds-DNA     circular SYN 29-JUL-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8789)
  AUTHORS   Stanford University
  TITLE     Direct Submission
  JOURNAL   Exported Jul 29, 2019 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8789
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    263..826
                     /label=pU6
                     /label=pU6 (pMJ177) 5' CMV promoter IE
     misc_feature    complement(766..775)
                     /label=GC Box
     misc_feature    844..1024
                     /label=HIV-1_5_LTR
     misc_feature    complement(1599..1605)
                     /label=TATA box
     misc_feature    1693..1926
                     /label=RRE
     misc_feature    2457..2471
                     /label=cPPT
     misc_feature    2680..3641
                     /label=EF1Alpha
     misc_feature    3643..3797
                     /label=EF1Alpha
                     /label=EF1Alpha(1)
     primer_bind     3804..3823
                     /label=oEJ288
     CDS             3863..4654
                     /codon_start=1
                     /label=G418
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    4655..4729
                     /label=T2A
     misc_feature    4655..4717
                     /label=puro-t2A
     misc_feature    4730..5488
                     /label=C1ORF43
     misc_feature    4933
                     /label=PAM mutation
     misc_feature    5489..5500
                     /label=linker
     CDS             5501..5566
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_recomb     5589..5622
                     /label=LoxP recombination site
                     /label=LoxP recombination site(1)
     misc_feature    5668..6268
                     /label=wPRE
     misc_feature    6458..6472
                     /label=cPPT
                     /label=cPPT(1)
     misc_feature    6795..6975
                     /label=HIV-1_5_LTR
                     /label=HIV-1_5_LTR(1)
     rep_origin      complement(7019..7701)
                     /direction=LEFT
                     /label=ColE1 origin
     misc_feature    complement(7022..7641)
                     /label=pBR322_origin
     misc_feature    complement(7046..7055)
                     /label=GC Box
                     /label=GC Box(1)
     CDS             complement(7799..8458)
                     /label=AmpR
     misc_feature    complement(8698..8726)
                     /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 ccgctgatct tcagacctgg
     1561 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat
     1621 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag
     1681 agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg
     1741 cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca
     1801 gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg
     1861 gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca
     1921 gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa
     1981 tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg
     2041 ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa
     2101 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa
     2161 ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg
     2221 cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg
     2281 atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga
     2341 aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg
     2401 atcggcactg cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa
     2461 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     2521 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     2581 acagggacag cagagatcca gtttggttag taccgggccc gcttcgagta ctaggatcca
     2641 ttaggcggcc gcgtggataa ccgtattacc gccatgcatg tgcccgtcag tgggcagagc
     2701 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct
     2761 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
     2821 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
     2881 acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct
     2941 ttacgggtta tggcccttgc gtgccttgaa ttacttccac ctggctgcag tacgtgattc
     3001 ttgatcccga gcttcgggtt ggaagtgggt gggagagttc gaggccttgc gcttaaggag
     3061 ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa
     3121 tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt
     3181 tttgatgacc tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag
     3241 atctgcacac tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc
     3301 agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt
     3361 agtctcaagc tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc
     3421 cctgggcggc aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc
     3481 ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg
     3541 agtcacccac acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca
     3601 cggagtaccg ggcgccgtcc aggcacctcg attagttctc gcgcttttgg agtacgtcgt
     3661 ctttaggttg gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga
     3721 ctgaagttag gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt
     3781 ttggatcttg gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc
     3841 aggtgtcgtg agctagccca ccatgattga acaagatgga ttgcacgcag gttctccggc
     3901 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3961 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4021 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4081 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4141 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4201 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4261 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4321 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4381 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4441 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4501 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4561 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4621 catcgccttc tatcgccttc ttgacgagtt cttcggatcg ggagagggca gaggaagtct
     4681 gctaacatgc ggtgacgtcg aggagaatcc tggcccaccg gtcgccacca tggcgtccgg
     4741 cagtaactgg ctctccgggg tgaatgtcgt gctggtgatg gcctacggga gcctggtgtt
     4801 tgtactgcta tttatttttg tgaagaggca aatcatgcgc tttgcaatga aatctcgaag
     4861 gggacctcat gtccctgtgg gacacaatgc ccccaaggac ttgaaagagg agattgatat
     4921 tcgactctcc agagttcagg atatcaagta tgagccccag ctccttgcag atgatgatgc
     4981 tagactacta caactggaaa cccagggaaa tcaaagttgc tacaactatc tgtataggat
     5041 gaaagctctg gatgccattc gtacctctga gatcccattt cattctgaag gccggcatcc
     5101 ccgttcctta atgggcaaga atttccgctc ctacctgctg gatctgcgaa acactagtac
     5161 gcctttcaag ggtgtacgca aagcactcat tgataccctt ttggatggct atgaaacagc
     5221 ccgctatggg acaggggtct ttggccagaa tgagtaccta cgctatcagg aggccctgag
     5281 tgagctggcc actgcggtta aagcacgaat tgggagctct cagcgacatc accagtcagc
     5341 agccaaagac ctaactcagt cccctgaggt ctccccaaca accatccagg tgacatacct
     5401 cccctccagt cagaagagta aacgtgccaa gcacttcctt gaattgaaga gctttaagga
     5461 taactataac acattggaga gtactctggg tggaagtggt gactacaaag accatgacgg
     5521 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtaag aattcgtcga
     5581 gggacctaat aacttcgtat agcatacatt atacgaagtt atacatgttt aagggttccg
     5641 gttccactag gtacaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     5701 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     5761 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     5821 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     5881 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     5941 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     6001 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     6061 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     6121 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     6181 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     6241 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgatcga gacctagaaa
     6301 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag
     6361 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa
     6421 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     6481 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag
     6541 gctacttccc tgattggcag aactacacac cagggccagg gatcagatat ccactgacct
     6601 ttggatggtg ctacaagcta gtaccagttg agcaagagaa ggtagaagaa gccaatgaag
     6661 gagagaacac ccgcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag
     6721 aagtattaga gtggaggttt gacagccgcc tagcatttca tcacatggcc cgagagctgc
     6781 atccggactg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     6841 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     6901 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     6961 ggaaaatctc tagcagcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     7021 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     7081 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     7141 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7201 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     7261 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7321 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7381 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7441 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7501 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7561 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7621 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7681 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7741 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7801 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7861 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7921 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7981 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     8041 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     8101 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     8161 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     8221 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     8281 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     8341 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8401 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8461 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8521 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8581 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8641 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8701 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8761 gcacatttcc ccgaaaagtg ccacctgac
//
