LOCUS       sequence-193524-        3715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CCR5-targeting plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3715)
  AUTHORS   Chen X, Janssen JM, Liu J, Maggio I, 't Jong AEJ, Mikkers HMM, 
            Goncalves MAFV
  TITLE     In trans paired nicking triggers seamless genome editing without 
            double-stranded DNA cutting.
  JOURNAL   Nat Commun. 2017 Sep 22;8(1):657. doi: 10.1038/s41467-017-00687-1.
  PUBMED    28939824
REFERENCE   2  (bases 1 to 3715)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2017-09-22"; volume: "8(1):657. doi"; pages: " 
            10.1038/s41467-017-00687-1"
COMMENT     Sequence Label: AY10_pS.Donor.R5.TS
FEATURES             Location/Qualifiers
     source          1..3715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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)."
     primer_bind     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    1049..1060
                     /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)"
     primer_bind     complement(1214..1231)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1214..1230)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1223..1245)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1439..1458)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1558..1580
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1618..1636)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1704..1808
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1809..1877,1878..2669)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2027..2046)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2840..3428
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3329..3348
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3582..3599
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgccaag cttgcatgca ggcctctgca gtcgacgggc ccgggatccg atacgacgcg
      181 gccgcagatg gtgctttcat gaattccccc aacagagcca agctctccat ctagtggaca
      241 gggaagctag cagcaaaatt taaatgcatt ctaggaggct gaggcaggag gatcgcttga
      301 gcccaggagt tcgagaccag cctgggcaac atagtgtgat cttgtatcta taaaaataaa
      361 caaaattagc ttggtgtggt ggcgcctgta gtccccagcc acttggaggg gtgaggtgag
      421 aggattgctt gagcccggga tggtccaggc tgcagtgagc catgatcgtg ccactgcact
      481 ccagcctggg cgacagagtg agaccctgtc tcacaacaac aacaacaaca acaaaaaggc
      541 tgagctgcac catgcttgac ccagtttctt aaaattgttg tcaaagcttc attcactcca
      601 tggtgctata gagcacaaga ttttatttgg tgagatggtg ctttcatgaa ttcccccaac
      661 agagcacgcg tgttaacaag cttctccatc tagtggacag ggaagctagc agcaaacctt
      721 cccttcacta caaaacttca ttgcttggcc aaaaagagag ttaattcaat gtagacatct
      781 atgtaggcaa ttaaaaacct attgatgtat aaaacagttt gcattcatgg agggcaacta
      841 aatacattct aggactttat aaaagatcac tttttattta tgcacagggt ggaacaagat
      901 ggattatcaa gtgtcaagtc caatctatga catcaattat tatacatcgg agccctgcca
      961 aaaaatcaat gtgaagcaaa tcgcagcccg cctcctgcct ccgctctact cactggtgtt
     1021 catctttggt tttgtgggca acatgctggt catcctcatc ctgataaact gcaaaaggct
     1081 gaagtttaaa cagatggtgc tttcatgaat tcccccaaca gagccaagct ctccatctag
     1141 tggacaggga agctagcagc aaacgccggc ggtcgtcatc tagatgcatt cgcgaggtac
     1201 cgagctcgaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     1261 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     1321 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc
     1381 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta
     1441 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     1501 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     1561 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     1621 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     1681 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1741 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     1801 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     1861 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     1921 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     1981 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     2041 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     2101 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     2161 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     2221 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     2281 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     2341 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     2401 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     2461 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     2521 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     2581 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     2641 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     2701 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     2761 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     2821 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2881 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2941 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     3001 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3061 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3121 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3181 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3241 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3301 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3361 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3421 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3481 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     3541 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     3601 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     3661 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaat
//
