LOCUS       Exported                7507 bp ds-DNA     circular SYN 31-OCT-2019
DEFINITION  Xm-0183.
ACCESSION   .
VERSION     .
KEYWORDS    pCAG_Xph15-mNeonGreen-CCR5TC
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7507)
  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..7507
                     /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..2073
                     /codon_start=1
                     /product="Paralog-specific PSD-95 binder Xph15 (Rimbault et
                     al., 2019)"
                     /label=Xph15
                     /translation="SSVSSVPTKLEVVAATPTSLLISWDAGPRNVSYYRITYGETGGNS
                     PVQEFTVPGSSSTATISGLSPGVDYTITVYASGHVSTLMTPISINYRT"
     CDS             2104..2808
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="VSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYEE
                     LNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFEDG
                     ASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIST
                     FKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQK
                     AFTDVMGMDELYK"
     regulatory      2356..2365
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2869..3210
                     /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(2915..2934)
                     /label=Xos-0059 pCAG-rev
     CDS             3259..3384
                     /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          3433..3565
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3574..4162
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3627..3647)
                     /label=WPRE-R
                     /note="From Addgene"
     CDS             complement(4045..4056)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4195..4306
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    4598..4652
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5011..5027)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5035..5051
                     /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(5059..5089)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5104..5125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5183..5379
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        5184..5379
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5230..5365
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5385..5519
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(5758..6346)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6815..7192)
                     /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(7377..7481)
                     /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 cagggcctcg taatgtcagt tattaccgta tcacctatgg cgaaacgggc
     1921 ggtaactccc cggttcagga attcaccgtc ccgggtagtt cctcaaccgc aacgatttcc
     1981 ggcctgtcac cgggtgtcga ctataccatt acggtttacg cttctggtca tgttagtacg
     2041 ctgatgacgc cgatttctat caattaccgt accggtagcg gctccagtag atctggtagc
     2101 ggcgtgagca agggcgagga ggataacatg gcctctctcc cagcgacaca tgagttacac
     2161 atctttggct ccatcaacgg tgtggacttt gacatggtgg gtcagggcac cggcaatcca
     2221 aatgatggtt atgaggagtt aaacctgaag tccaccaagg gtgacctcca gttctccccc
     2281 tggattctgg tccctcatat cgggtatggc ttccatcagt acctgcccta ccctgacggg
     2341 atgtcgcctt tccaggccgc catggtagat ggctccggat accaagtcca tcgcacaatg
     2401 cagtttgaag atggtgcctc ccttactgtt aactaccgct acacctacga gggaagccac
     2461 atcaaaggag aggcccaggt gaaggggact ggtttccctg ctgacggtcc tgtgatgacc
     2521 aactcgctga ccgctgcgga ctggtgcagg tcgaagaaga cttaccccaa cgacaaaacc
     2581 atcatcagta cctttaagtg gagttacacc actggaaatg gcaagcgcta ccggagcact
     2641 gcgcggacca cctacacctt tgccaagcca atggcggcta actatctgaa gaaccagccg
     2701 atgtacgtgt tccgtaagac ggagctcaag cactccaaga ccgagctcaa cttcaaggag
     2761 tggcaaaagg cctttaccga tgtgatgggc atggacgagc tgtacaaggg tagcggctcc
     2821 tctgctagcg gagctggcgc tggagcgggt gcaggggctg gctctagatt ccagtgccgg
     2881 atctgcatgc ggaacttcag cgaccggtcc aacctgagca ggcacatcag aacccacacc
     2941 ggagaaaagc ccttcgcctg cgacatttgc ggccggaagt tcgccatcag cagcaacctg
     3001 aacagccaca ccaagatcca cactggcagc cagaaacctt tccagtgcag aatttgtatg
     3061 agaaacttta gcagaagcga caacctggcc agacacatcc ggacacatac tggtgaaaaa
     3121 ccttttgcct gtgatatctg tggcagaaag tttgccacct ccggcaatct gacccggcac
     3181 acaaagattc acctgcgggg cagccagcta tcgattgtcg acgctcctga acaacgtgaa
     3241 ggtgcttctc aagtttctgt ttctgttact tttgaagatg ttgctgttct ttttactcgt
     3301 gatgaatgga aaaaacttga tctttctcaa cgttctcttt atcgtgaagt tatgcttgaa
     3361 aattattcta atcttgcttc tatggcttaa acgcgttgta caagtaaagc ggccccggac
     3421 tcgaggccgc aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg
     3481 ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac
     3541 atccactttg cctttctctc cacaggtgtc gacaatcaac ctctggatta caaaatttgt
     3601 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3661 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     3721 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     3781 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     3841 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     3901 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     3961 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     4021 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     4081 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     4141 tccctttggg ccgcctcccc gcctggaatt cgagctcggt acgatcagcc tcgactgtgc
     4201 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     4261 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     4321 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     4381 acaatagcag gccagctttt gttcccttta gggttaattg cgcgcttggc gctaatcatg
     4441 gtcatagctg tttccactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg
     4501 gccaatgccc tggctcacaa ataccactga gatcgttttc cctctgccaa aaattatggg
     4561 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt
     4621 gcaatagtgt gttgaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     4681 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc atatgctggc
     4741 tgccatgaac aaaggtggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc
     4801 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttta tattttgttt
     4861 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt
     4921 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg aagatccctc
     4981 gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5041 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5101 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5161 aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta
     5221 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     5281 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     5341 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag
     5401 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5461 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc
     5521 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5581 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5641 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5701 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5761 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5821 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5881 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5941 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6001 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6061 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6121 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6181 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6241 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6301 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6361 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6421 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6481 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6541 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6601 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6661 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6721 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6781 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6841 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6901 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6961 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggagc actgcataat
     7021 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7081 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7141 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7201 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7261 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7321 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7381 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7441 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7501 ccacctg
//
