LOCUS       sequence-388199-        7898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Loads HiBiT-tagged L fragment of a zinc finger nuclease specific for
            human AAVS1 locus using Pdp1 packaging domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7898)
  AUTHORS   Kreitz J, Friedrich MJ, Guru A, Lash B, Saito M, Macrae RK, Zhang F
  TITLE     Programmable protein delivery with a bacterial contractile injection
            system.
  JOURNAL   Nature. 2023 Apr;616(7956):357-364. doi: 10.1038/s41586-023-05870-7.
            Epub 2023 Mar 29.
  PUBMED    36991127
REFERENCE   2  (bases 1 to 7898)
  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"; doi: 
            "10.1038/s41586-023-05870-7"; journalName: "Nature"; date: 
            "2023-04"; volume: "616"; issue: "7956"; pages: "357-364"
COMMENT     Sequence Label: pBR322-Pdp1_NTD-ZFN_L-HiBiT
FEATURES             Location/Qualifiers
     source          1..7898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(207..311)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     379..397
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(424..429,430..446,447..452)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             1372..1392
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5866..6057
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6051..6073)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6159..6299
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6214..6233
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6314..6331)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6485..7073)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6565..6584)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7244..7898,1..137,138..206))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7867..7886
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
       61 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
      121 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
      181 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
      241 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
      301 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
      361 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca agaattctca
      421 tgtttgacag cttatcatcg ataagcttta atgcggtagt ttatcacagt taaattgcta
      481 acgcagtcag gcaccgtgta tgaaatctaa caatgcgctc atcgtcatcc tcggcaccgt
      541 caccctggat gctgtaggca taggcttggt tatgccggta ctgccgggcc tcttgcggga
      601 tatcgtccat tccgacagca tcgccagtca ctatggcgtg ctgctagcgc tatatgcgtt
      661 gatgcaattt ctatgcgcac ccgttctcgg agcactgtcc gaccgctttg gccgccgccc
      721 agtcctgctc gcttcgctac ttggagccac tatcgactac gcgatcatgg cgaccacacc
      781 cgtcctgtgg atccattttt ttaattaaat aaaggaatta atattgattt gatagagcat
      841 gtaatcaata acttctatga aaatgttgaa aaacaataca cggaaacact taacttacaa
      901 ttaaattgtt aaaggagatg atgataagaa tggtcagtca ccagataaaa tgatctctgg
      961 tcaattttag gaaagctgaa aaaacctgac caaatgcgtc gtaaattcta tgcttagaaa
     1021 gactcggtgc cagtgtaata cctaagatat ggtttttatt tccatatctg ttcgaaaact
     1081 tagaaataac aagttcaaca ataaccaggg actttaaata tgcctagata tgctaattat
     1141 cagataaacc ccaaacagaa tattaaaaat tcacatggga aatcttcatc gtcagatttt
     1201 tctagtgggt acctttcatt ctcaaataat tcgcttgatg acccttttat tcggcagcaa
     1261 gttaagagag aatttatttg ggagggacat atgaaggaga ttgaagaagc ttcaagatta
     1321 ggtggctctg gtggtggcgg ttctggcggc ggtggttctg gcggtatggc accgaaaaag
     1381 aaacgcaaag ttggtattca tggtgttccg gcagcaatgg cagaacgtcc gtttcagtgt
     1441 cgtatttgta tgcgcaattt tagctataac tggcatctgc agcgtcatat tcgtacccat
     1501 accggtgaaa aaccgtttgc atgtgatatt tgcggtcgta aatttgcacg tagcgatcat
     1561 ctgaccacac ataccaaaat tcataccggt agccagaaac cttttcagtg ccgcatatgc
     1621 atgcgtaact ttagtcataa ctatgcccgt gattgccata ttcgcacaca cacaggcgaa
     1681 aaacctttcg cctgcgatat ctgtggacgt aaattcgcac agaatagcac ccgtattggt
     1741 catacaaaaa tccatctgcg tggtagccag ctggttaaaa gcgaactgga agaaaagaaa
     1801 agtgaactgc gccacaaact gaaatatgtg ccgcatgaat atatcgagct gattgaaatt
     1861 gcccgtaata gtacccagga tcgtattctg gaaatgaagg tgatggaatt ctttatgaaa
     1921 gtgtatggct atcgcggtaa acatttaggt ggtagccgta aaccggatgg tgcaatttat
     1981 accgttggta gcccgattga ttatggtgtt attgttgata ccaaagccta tagcggtggt
     2041 tataatctgc cgattggtca ggcagatgaa atggaacgtt atgtcgaaga aaatcagacc
     2101 cgcaataaac atctgaatcc gaatgaatgg tggaaagttt atccgagcag cgttaccgaa
     2161 ttcaaatttc tgtttgtgag cggtcacttc aaaggcaact ataaagcaca gctgacccgt
     2221 ctgaatcata ttaccaattg taatggtgca gttctgagcg ttgaagaact gctgattggt
     2281 ggtgaaatga ttaaagcagg tacactgacc ctggaagaag ttcgtcgcaa atttaacaat
     2341 ggcgagatca actttgggag ctccggtgtg agcggctggc ggctgttcaa gaagattagc
     2401 taatatggat tttcatgtca tcaggagaat attcaaaatt ctgtgtcaca caaaatattt
     2461 tacagcgtaa tcacaccatt aagaggggca tcaaaataaa tggttaactg ggacaatgtt
     2521 gaatttcatt tttataaggc attgtccgta ttatccacca tccagatcaa cggattcaga
     2581 tggtgtaaag aaagcatata gctggtaggt aaaattaaga agtgtagttt tataaagtat
     2641 taatggagaa cgatgaattt aaagtaaatg gaggaaggtt agtttcacta tcaaattaca
     2701 ggaccaaaaa tatggagtat tccatcttct ttggtgaatt aagtcttgaa gacttcaaca
     2761 aatattgcct ggaactatgg gcctgtgaat ggtattggag tatcatttat aacgactatt
     2821 tcgataggaa atttaaagtc ttgttgaaaa tgactaatcg tctaaaagta tgtctggaga
     2881 aataggttta atctctatat ttaacggttt tattttcatc tatttactaa tggtaaattt
     2941 ttttactctt tgaatatatt ggtgagtttg ttatccccat tagttaccag ggtattatta
     3001 atgagtgaaa ttcaatttta ttgatattaa agtcaaattt acaaatgatg actttattat
     3061 tttttattta ttaattttaa attatttttt gttaatcttt actataaatg aattttttgc
     3121 ttgcatggac ttgatttttt tatatataac attgacaaat attaattcat atatatgatt
     3181 gttatagtga gttatttaat tgattcgact aacaatttca ttgtaatttt tcaagatagc
     3241 agtcaattgc tataacaagc atctttctcg tcaaaggaaa atgtgtatgg cctataagca
     3301 gcaacttgaa agatgaagga tataatgatg attaacttaa cggagagtag atattagaag
     3361 tagtagaagg acaatttaag actttatacc cattaaaaca aaggggtaaa ataatgatgt
     3421 ttgatgaaaa tttagaatgt gttaatcacg aaattatcaa tgaaaaatta gatacgaaaa
     3481 ttcaccttgc taatccttat gccttttggg ggggaagggt aaatgagaat ataaatgggt
     3541 taattaaaca acattcttta agaggaactg actttaataa aatttttaac cggaagataa
     3601 atttttttgt aaatggaata aataattgcc cccgaaaaac tcggaatata aaacccccta
     3661 attaattatt taaaggaata agagtttatt tgctcgtaga gtaattatgt tccattaata
     3721 tgctaaattg ctcagtaaag ttattaactt tttttcaacg ccagtgaata agtctgtttt
     3781 attaaataag gagttcaggt gtttatttca aaagaattat caagctttat tgctgtagct
     3841 aaaaataaat caataaataa agcaacaacg gaacttttta ttacacagtc accgatttgc
     3901 cgaagcttaa aaaaactgga attttcatta ggtgtaaaac tctttgtaag aaaaagccag
     3961 ggattggctt taacagaaga aggagaatgc ttatataata aagttttgtc aatttgtgaa
     4021 cggcttcaag aaatagaagg tgactacaag aatgaaaaat caacatacaa tgatctatat
     4081 caaaaaaata tatgaggttc taatataata agcggttcta gataactaaa gtcatctatt
     4141 accaatatct gtatactcag tgaaagtttt gtcttctagg taagttttca taatagtgac
     4201 ataccttatt ttttcatctg ggtgtctctc atcctgaaga atgggtgata tagttgaaat
     4261 aactcagttc gttttgtggt ttacgatttt tggcggatca caaaaatagg gctgagtccc
     4321 ttcaagtgag ttttgaaata tacaattata cccttcatcc tttaagttgt tgctttgttg
     4381 gctgttttca ttcacctcag tcacatagtt atctatgctc ctgaggattc gttcacttgc
     4441 cgtcgtgctg caactcgaaa tctattgggt atatatgtaa ttttcatatt ctgtttcatt
     4501 ttaacggtat tatttaaata tttatattta gaacattagg ttattagaat agataatttt
     4561 aggtaaatgt aattaattca accatcacgg agagtttatc agtcgaccga tgcccttgag
     4621 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
     4681 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat
     4741 tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt
     4801 cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca aacgtttcgg
     4861 cgagaagcag gccattatcg ccggcatggc ggccgacgcg ctgggctacg tcttgctggc
     4921 gttcgcgacg cgaggctgga tggccttccc cattatgatt cttctcgctt ccggcggcat
     4981 cgggatgccc gcgttgcagg ccatgctgtc caggcaggta gatgacgacc atcagggaca
     5041 gcttcaagga tcgctcgcgg ctcttaccag cctaacttcg atcattggac cgctgatcgt
     5101 cacggcgatt tatgccgcct cggcgagcac atggaacggg ttggcatgga ttgtaggcgc
     5161 cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc
     5221 gacctgaatg gaagccggcg gcacctcgct aacggattca ccactccaag aattggagcc
     5281 aatcaattct tgcggagaac tgtgaatgcg caaaccaacc cttggcagaa catatccatc
     5341 gcgtccgcca tctccagcag ccgcacgcgg cgcatctcgg gcagcgttgg gtcctggcca
     5401 cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt
     5461 actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa
     5521 acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg
     5581 gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc
     5641 tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg
     5701 atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt
     5761 aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt
     5821 atcattaccc ccatgaacag agatccccct tacacggagg catcagtgac caaacaggaa
     5881 aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa
     5941 ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct
     6001 gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     6061 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     6121 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     6181 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
     6241 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     6301 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6361 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6421 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6481 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6541 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6601 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6661 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6721 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6781 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6841 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6901 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     6961 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7021 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7081 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7141 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaacgaagtt
     7201 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7261 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7321 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7381 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7441 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7501 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgctg
     7561 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7621 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7681 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7741 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7801 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7861 cacgggataa taccgcgcca catagcagaa ctttaaaa
//
