LOCUS       sequence-193523-        3559 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 3559)
  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 3559)
  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: AY42_pS.Donor.R5
FEATURES             Location/Qualifiers
     source          1..3559
                     /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    971..982
                     /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(1058..1075)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1058..1074)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1067..1089)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1283..1302)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1402..1424
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1462..1480)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1548..1652
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1653..1721,1722..2513)
                     /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(1871..1890)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2684..3272
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3173..3192
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3426..3443
                     /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 gccaaatgca ttctaggagg ctgaggcagg aggatcgctt gagcccagga gttcgagacc
      241 agcctgggca acatagtgtg atcttgtatc tataaaaata aacaaaatta gcttggtgtg
      301 gtggcgcctg tagtccccag ccacttggag gggtgaggtg agaggattgc ttgagcccgg
      361 gatggtccag gctgcagtga gccatgatcg tgccactgca ctccagcctg ggcgacagag
      421 tgagaccctg tctcacaaca acaacaacaa caacaaaaag gctgagctgc accatgcttg
      481 acccagtttc ttaaaattgt tgtcaaagct tcattcactc catggtgcta tagagcacaa
      541 gattttattt ggtgagatgg tgctttcatg aattccccca acagagcacg cgtgttaaca
      601 agcttctcca tctagtggac agggaagcta gcagcaaacc ttcccttcac tacaaaactt
      661 cattgcttgg ccaaaaagag agttaattca atgtagacat ctatgtaggc aattaaaaac
      721 ctattgatgt ataaaacagt ttgcattcat ggagggcaac taaatacatt ctaggacttt
      781 ataaaagatc actttttatt tatgcacagg gtggaacaag atggattatc aagtgtcaag
      841 tccaatctat gacatcaatt attatacatc ggagccctgc caaaaaatca atgtgaagca
      901 aatcgcagcc cgcctcctgc ctccgctcta ctcactggtg ttcatctttg gttttgtggg
      961 caacatgctg gtcatcctca tcctgataaa ctgcaaaagg ctgaagtttc cggcggtcgt
     1021 catctagatg cattcgcgag gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc
     1081 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     1141 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     1201 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     1261 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     1321 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     1381 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     1441 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     1501 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1561 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1621 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1681 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1741 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1801 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     1861 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     1921 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     1981 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2041 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2101 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2161 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     2221 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2281 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2341 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2401 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2461 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2521 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     2581 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     2641 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2701 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2761 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2821 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     2881 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2941 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3001 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3061 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3121 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3181 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3241 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3301 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3361 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3421 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     3481 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     3541 gcagtgagcg caacgcaat
//
