LOCUS       sequence-183601-        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"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    549..560
                     /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(1494..1510)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1494..1510)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1507..1529)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1518..1534
                     /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(join(1542..1548,1549..1566,1567..1572))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1587..1608
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1725..1742)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1896..2484)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1976..1995)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2655..3446,3447..3515))
                     /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     3278..3297
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3516..3620)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3688..3706
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
      421 tgcatctaga tgacgaccgc cggcgtttgc tgctagcttc cctgtccact agatggagag
      481 cttggctctg ttgggggaat tcatgaaagc accatctgtt taaacttcag ccttttgcag
      541 tttatcagga tgaggatgac cagcatgttg cccacaaaac caaagatgaa caccagtgag
      601 tagagcggag gcaggaggcg ggctgcgatt tgcttcacat tgattttttg gcagggctcc
      661 gatgtataat aattgatgtc atagattgga cttgacactt gataatccat cttgttccac
      721 cctgtgcata aataaaaagt gatcttttat aaagtcctag aatgtattta gttgccctcc
      781 atgaatgcaa actgttttat acatcaatag gtttttaatt gcctacatag atgtctacat
      841 tgaattaact ctctttttgg ccaagcaatg aagttttgta gtgaagggaa ggtttgctgc
      901 tagcttccct gtccactaga tggagaagct tgttaacacg cgtgctctgt tgggggaatt
      961 catgaaagca ccatctcacc aaataaaatc ttgtgctcta tagcaccatg gagtgaatga
     1021 agctttgaca acaattttaa gaaactgggt caagcatggt gcagctcagc ctttttgttg
     1081 ttgttgttgt tgttgtgaga cagggtctca ctctgtcgcc caggctggag tgcagtggca
     1141 cgatcatggc tcactgcagc ctggaccatc ccgggctcaa gcaatcctct cacctcaccc
     1201 ctccaagtgg ctggggacta caggcgccac cacaccaagc taattttgtt tatttttata
     1261 gatacaagat cacactatgt tgcccaggct ggtctcgaac tcctgggctc aagcgatcct
     1321 cctgcctcag cctcctagaa tgcatttaaa ttttgctgct agcttccctg tccactagat
     1381 ggagagcttg gctctgttgg gggaattcat gaaagcacca tctgcggccg cgtcgtatcg
     1441 gatcccgggc ccgtcgactg cagaggcctg catgcaagct tggcgtaatc atggtcatag
     1501 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     1561 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     1621 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     1681 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     1741 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1801 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     1861 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     1921 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     1981 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2041 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2101 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2161 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2221 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2281 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2341 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2401 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2461 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2521 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2581 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2641 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     2701 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     2761 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     2821 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     2881 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     2941 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3001 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3061 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3121 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3181 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3241 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     3301 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     3361 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     3421 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     3481 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     3541 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     3601 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     3661 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtc
//
