LOCUS       Exported                7516 bp ds-DNA     circular SYN 31-OCT-2019
DEFINITION  Xm-0176.
ACCESSION   .
VERSION     .
KEYWORDS    pCAG_Xph20-mRuby2-CCR5TC
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7516)
  AUTHORS   Matthieu Sainlos
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Oct 31, 2019 from SnapGene 5.0.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7516
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    24..35
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     enhancer        39..418
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        421..696
                     /label=chicken beta-actin promoter
     intron          697..1708
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1716..1735
                     /label=pCAG-F
                     /note="From Addgene list"
     regulatory      1780..1789
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1795..2079
                     /codon_start=1
                     /product="Paralog-specific PSD-95 binder Xph20 (Rimbault et
                     al., 2019)"
                     /label=Xph20
                     /translation="SSVSSVPTKLEVVAATPTSLLISWDAVAKNVKVGYYRITYGETGG
                     NSPVQEFTVPGSSSTATISGLSPGVDYTITVYANGVLSKMVLPISINYRT"
     CDS             2110..2817
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTHM
                     ALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAVA
                     KFAGLGGGMDELYK"
     CDS             2878..3219
                     /codon_start=1
                     /product="zinc-finger domain from the CCR5-224 (+) 
                     zinc-finger nuclease that recognizes the left half-site 
                     target sequence in the human CCR5 gene (Perez et al., 2008;
                     Gross et al., 2013)"
                     /label=CCR5TC
                     /translation="FQCRICMRNFSDRSNLSRHIRTHTGEKPFACDICGRKFAISSNLN
                     SHTKIHTGSQKPFQCRICMRNFSRSDNLARHIRTHTGEKPFACDICGRKFATSGNLTRH
                     TKIHLRGSQL"
     primer_bind     complement(2924..2943)
                     /label=Xos-0059 pCAG-rev
     CDS             3268..3393
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the rat zinc finger protein Kid-1 
                     (Witzgall et al., 1994)"
                     /label=KRAB-A
                     /translation="VSVTFEDVAVLFTRDEWKKLDLSQRSLYREVMLENYSNLASM"
     intron          3442..3574
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3583..4171
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3636..3656)
                     /label=WPRE-R
                     /note="From Addgene"
     CDS             complement(4054..4065)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4204..4315
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    4607..4661
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5020..5036)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5044..5060
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5068..5098)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5113..5134
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5192..5388
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        5193..5388
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5239..5374
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5394..5528
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(5767..6355)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6824..7201)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     promoter        complement(7386..7490)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agagtcatcc tcatcgtcga cattgattat tgactagtta
       61 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      241 ggactattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggg
      421 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
      481 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg
      541 cgcgcgccag gcggggcggg gcgggggcga gggcggggcg gggcgaggcg gagaggtgcg
      601 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
      661 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgtt gccttcgccc
      721 cgtgccccgc tccgcgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
      781 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa
      841 tgacggctcg tttcttttct gtggctgcgt gaaagcctta aagggctccg ggagggccct
      901 ttgtgcgggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc
      961 gtgcggcccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg
     1021 ctccgcgtgt gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc gggggggctg
     1081 cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg
     1141 cgcggcggtc gggctgtaac ccccccctgc acccccctcc ccgagttgct gagcacggcc
     1201 cggcttcggg tgcggggctc cgtgcggggc gtggcgcggg gctcgccgtg ccgggcgggg
     1261 ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg gagggctcgg
     1321 gggaggggcg cggcggcccc ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca
     1381 ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc aaatctgtgc
     1441 ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg cgaagcggtg
     1501 cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc gccgccgtcc
     1561 ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc gggggggacg
     1621 gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca
     1681 tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc
     1741 tcatcatttt ggcaaagaat tcggtaccgc gggcccgggg ccaccatggc gggcagctct
     1801 gtcagttccg tgccgaccaa actggaagtg gttgcggcca ccccgacgtc tctgctgatc
     1861 agctgggatg cagtggcgaa gaatgtgaag gtcggttatt accgtatcac ctatggcgaa
     1921 acgggcggta actccccggt tcaggaattc accgtcccgg gtagttcctc aaccgcaacg
     1981 atttccggcc tgtcaccggg tgtcgactat accattacgg tttacgctaa tggggtgctg
     2041 agtaagatgg tgcttccgat ttctatcaat taccgtaccg gtagcggctc cagtagatct
     2101 ggtagcggcg tgtctaaggg cgaagagctg atcaaggaaa atatgcgtat gaaggtggtc
     2161 atggaaggtt cggtcaacgg ccaccaattc aaatgcacag gtgaaggaga aggcaatccg
     2221 tacatgggaa ctcaaaccat gaggatcaaa gtcatcgagg gaggacccct gccatttgcc
     2281 tttgacattc ttgccacgtc gttcatgtat ggcagccgta cttttatcaa gtacccgaaa
     2341 ggcattcctg atttctttaa acagtccttt cctgagggtt ttacttggga aagagttacg
     2401 agatacgaag atggtggagt cgtcaccgtc atgcaggaca ccagccttga ggatggctgt
     2461 ctcgtttacc acgtccaagt cagaggggta aactttccct ccaatggtcc cgtgatgcag
     2521 aagaagacca agggttggga gcctaataca gagatgatgt atccagcaga tggtggtctg
     2581 aggggataca ctcatatggc actgaaagtt gatggtggtg gccatctgtc ttgctctttc
     2641 gtaacaactt acaggtcaaa aaagaccgtc gggaacatca agatgcccgg tatccatgcc
     2701 gttgatcacc gcctggaaag gttagaggaa agtgacaatg aaatgttcgt agtacaacgc
     2761 gaacacgcag ttgccaagtt cgccgggctt ggtggtggga tggacgagct gtacaagggt
     2821 agcggctcct ctgctagcgg agctggcgct ggagcgggtg caggggctgg ctctagattc
     2881 cagtgccgga tctgcatgcg gaacttcagc gaccggtcca acctgagcag gcacatcaga
     2941 acccacaccg gagaaaagcc cttcgcctgc gacatttgcg gccggaagtt cgccatcagc
     3001 agcaacctga acagccacac caagatccac actggcagcc agaaaccttt ccagtgcaga
     3061 atttgtatga gaaactttag cagaagcgac aacctggcca gacacatccg gacacatact
     3121 ggtgaaaaac cttttgcctg tgatatctgt ggcagaaagt ttgccacctc cggcaatctg
     3181 acccggcaca caaagattca cctgcggggc agccagctat cgattgtcga cgctcctgaa
     3241 caacgtgaag gtgcttctca agtttctgtt tctgttactt ttgaagatgt tgctgttctt
     3301 tttactcgtg atgaatggaa aaaacttgat ctttctcaac gttctcttta tcgtgaagtt
     3361 atgcttgaaa attattctaa tcttgcttct atggcttaaa cgcgttgtac aagtaaagcg
     3421 gccccggact cgaggccgca ggtaagtatc aaggttacaa gacaggttta aggagaccaa
     3481 tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc acctattggt
     3541 cttactgaca tccactttgc ctttctctcc acaggtgtcg acaatcaacc tctggattac
     3601 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     3661 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     3721 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     3781 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     3841 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     3901 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     3961 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg
     4021 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     4081 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     4141 agtcggatct ccctttgggc cgcctccccg cctggaattc gagctcggta cgatcagcct
     4201 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     4261 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     4321 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     4381 attgggaaga caatagcagg ccagcttttg ttccctttag ggttaattgc gcgcttggcg
     4441 ctaatcatgg tcatagctgt ttccactcct caggtgcagg ctgcctatca gaaggtggtg
     4501 gctggtgtgg ccaatgccct ggctcacaaa taccactgag atcgttttcc ctctgccaaa
     4561 aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt
     4621 attttcattg caatagtgtg ttgaattttt tgtgtctctc actcggaagg acatatggga
     4681 gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca acatatgcca
     4741 tatgctggct gccatgaaca aaggtggcta taaagaggtc atcagtatat gaaacagccc
     4801 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat ttttttttat
     4861 attttgtttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     4921 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatga
     4981 agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5041 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5101 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5161 ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc
     5221 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5281 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     5341 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt
     5401 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5461 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5521 tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5581 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5641 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5701 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5761 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5821 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5881 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5941 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6001 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6061 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6121 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6181 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6241 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6301 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6361 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6421 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6481 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6541 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6601 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6661 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6721 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6781 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6841 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6901 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6961 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggagca
     7021 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7081 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7141 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7201 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7261 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7321 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7381 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7441 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7501 cgaaaagtgc cacctg
//
