LOCUS       Exported                7462 bp ds-DNA     circular SYN 31-OCT-2019
DEFINITION  Xm-0110.
ACCESSION   .
VERSION     .
KEYWORDS    
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7462)
  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..7462
                     /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             2104..2778
                     /codon_start=1
                     /product="improved monomeric variant of green-to-red 
                     photoswitchable fluorescent protein EosFP (Zhang et al., 
                     2012)"
                     /label=mEos3.2
                     /translation="SAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVKE
                     GGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNARN
                     DITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIEMALLLE
                     GNAHYRCDFRTTYKAKEKGVKLPGAHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPDN
                     ARR"
     CDS             2824..3165
                     /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(2870..2889)
                     /label=Xos-0059 pCAG-rev
     CDS             3214..3339
                     /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          3388..3520
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3529..4117
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3582..3602)
                     /label=WPRE-R
                     /note="From Addgene"
     CDS             complement(4000..4011)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4150..4261
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    4553..4607
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4966..4982)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4990..5006
                     /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(5014..5044)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5059..5080
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5139..5334
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5185..5320
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5340..5474
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(5713..6301)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6770..7147)
                     /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(7332..7436)
                     /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 gggagtgcga ttaagccaga catgaagatc aaactccgta tggaaggcaa cgtaaacggg
     2161 caccactttg tgatcgacgg agatggtaca ggcaagcctt ttgagggaaa acagagtatg
     2221 gatcttgaag tcaaagaggg cggacctctg ccttttgcct ttgatatcct gaccactgca
     2281 ttccattacg gcaacagggt attcgccaaa tatccagaca acatacaaga ctattttaag
     2341 cagtcgtttc ctaaggggta ttcgtgggaa cgaagcttga ctttcgaaga cgggggcatt
     2401 tgcaacgcca gaaacgacat aacaatggaa ggggacactt tctataataa agttcgattt
     2461 tatggtacca actttcccgc caatggtcca gttatgcaga agaagacgct gaaatgggag
     2521 ccctccactg agaaaatgta tgtgcgtgat ggagtgctga cgggtgatat tgagatggct
     2581 ttgttgcttg aaggaaatgc ccattaccga tgtgacttca gaactactta caaagctaag
     2641 gagaagggtg tcaagttacc aggcgcccac tttgtggacc actgcattga gattttaagc
     2701 catgacaaag attacaacaa ggttaagctg tatgagcatg ctgttgctca ttctggattg
     2761 cctgacaatg ccagacgagc tagcggagct ggcgctggag cgggtgcagg ggctggctct
     2821 agattccagt gccggatctg catgcggaac ttcagcgacc ggtccaacct gagcaggcac
     2881 atcagaaccc acaccggaga aaagcccttc gcctgcgaca tttgcggccg gaagttcgcc
     2941 atcagcagca acctgaacag ccacaccaag atccacactg gcagccagaa acctttccag
     3001 tgcagaattt gtatgagaaa ctttagcaga agcgacaacc tggccagaca catccggaca
     3061 catactggtg aaaaaccttt tgcctgtgat atctgtggca gaaagtttgc cacctccggc
     3121 aatctgaccc ggcacacaaa gattcacctg cggggcagcc agctatcgat tgtcgacgct
     3181 cctgaacaac gtgaaggtgc ttctcaagtt tctgtttctg ttacttttga agatgttgct
     3241 gttcttttta ctcgtgatga atggaaaaaa cttgatcttt ctcaacgttc tctttatcgt
     3301 gaagttatgc ttgaaaatta ttctaatctt gcttctatgg cttaaacgcg ttgtacaagt
     3361 aaagcggccc cggactcgag gccgcaggta agtatcaagg ttacaagaca ggtttaagga
     3421 gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct
     3481 attggtctta ctgacatcca ctttgccttt ctctccacag gtgtcgacaa tcaacctctg
     3541 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     3601 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     3661 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     3721 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     3781 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     3841 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     3901 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     3961 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     4021 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     4081 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattcgagc tcggtacgat
     4141 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     4201 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     4261 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     4321 gggaggattg ggaagacaat agcaggccag cttttgttcc ctttagggtt aattgcgcgc
     4381 ttggcgctaa tcatggtcat agctgtttcc actcctcagg tgcaggctgc ctatcagaag
     4441 gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatcg ttttccctct
     4501 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
     4561 aaatttattt tcattgcaat agtgtgttga attttttgtg tctctcactc ggaaggacat
     4621 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     4681 atgccatatg ctggctgcca tgaacaaagg tggctataaa gaggtcatca gtatatgaaa
     4741 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt
     4801 ttttatattt tgtttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt
     4861 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc
     4921 ttatgaagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc
     4981 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5041 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5101 cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc
     5161 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     5221 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     5281 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta
     5341 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5401 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5461 atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5521 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5581 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5641 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5701 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5761 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5821 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5881 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     5941 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6001 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6061 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6121 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     6181 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6241 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6301 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6361 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6421 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6481 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6541 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6601 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6661 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6721 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6781 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6841 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6901 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6961 ggagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7021 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7081 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7141 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7201 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7261 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7321 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7381 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7441 tttccccgaa aagtgccacc tg
//
