LOCUS       sequence-388686-        8033 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For loading L half of zinc finger deaminase targeting human TRAC 
            locus (ZFD_TRAC-NC-Left-G1397-N_DddA); also contains PVC regulatory 
            genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8033)
  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 8033)
  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-ZFD_L
FEATURES             Location/Qualifiers
     source          1..8033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3466..3657
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3651..3673)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3759..3899
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3814..3833
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3914..3931)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4085..4673)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4165..4184)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4844..5635,5636..5704))
                     /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     5467..5486
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5705..5809)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5877..5895
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(5922..5927,5928..5944,5945..5950)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             6864..6887
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6897..6917
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7782..8033
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
ORIGIN
        1 taatatggat tttcatgtca tcaggagaat attcaaaatt ctgtgtcaca caaaatattt
       61 tacagcgtaa tcacaccatt aagaggggca tcaaaataaa tggttaactg ggacaatgtt
      121 gaatttcatt tttataaggc attgtccgta ttatccacca tccagatcaa cggattcaga
      181 tggtgtaaag aaagcatata gctggtaggt aaaattaaga agtgtagttt tataaagtat
      241 taatggagaa cgatgaattt aaagtaaatg gaggaaggtt agtttcacta tcaaattaca
      301 ggaccaaaaa tatggagtat tccatcttct ttggtgaatt aagtcttgga gacttcaaca
      361 aatattgcct ggaactatgg gcctgtgaat ggtattggag tatcatttat aacgactatt
      421 tcgataggaa atttaaagtc ttgttgaaaa tgactaatcg tctaaaagta tgtctggaga
      481 aataggttta atctctatac ttaacggttt tattttcatc tatttactaa tggtaaattt
      541 ttttactctt tgaatatatt ggtgagtttg ttatccccat tagttaccag ggtattatta
      601 atgagtgaaa ttcaatttta ttgatattaa agtcaaattt acaaatgatg actttattat
      661 tttttattta ttaattttaa attatttttt gttaatcttt actataaatg aattttttgc
      721 ttgcatggac ttgatttttt tatatataac attgacaaat attaattcat atatatgatt
      781 gttatagtga gttatttaat tgattcgact aacaatttca ttgtaatttt tcaagatagc
      841 agtcaattgc tataacaagc atctttctcg tcaaaggaaa atgtgtatgg cctataagca
      901 gcaacttgaa agatgaagga tataatgatg attaacttaa cggagagtag atattagaag
      961 tagtagaagg acaatttaag actttatacc cattaaaaca aaggggtaaa ataatgatgt
     1021 ttgatgaaaa tttagaatgt gttaatcacg aaattatcaa tgaaaaatta gatacgaaaa
     1081 ttcaccttgc taatccttat gccttttggg ggggaagggt aaatgagaat ataaatgggt
     1141 taattaaaca acattcttta agaggaactg actttaataa aatttttaac cggaagataa
     1201 atttttttgt aaatggaata aataattgcc cccgaaaaac tcggaatata aaacccccta
     1261 attaattatt taaaggaata agagtttatt tgctcgtaga gtaattatgt tccattaata
     1321 tgctaaattg ctcagtaaag ttattaactt tttttcaacg ccagtgaata agtctgtttt
     1381 attaaataag gagttcaggt gtttatttca aaagaattat caagctttat tgctgtagct
     1441 aaaaataaat caataaataa agcaacaacg gaacttttta ttacacagtc accgatttgc
     1501 cgaagcttaa aaaaactgga attttcatta ggtgtaaaac tctttgtaag aaaaagccag
     1561 ggattggctt taacagaaga aggagaatgc ttatataata aagttttgtc aatttgtgaa
     1621 cggcttcaag aaatagaagg tgactacaag aatgaaaaat caacatacaa tgatctatat
     1681 caaaaaaata tatgaggttc taatataata agcggttcta gataactaaa gtcatctatc
     1741 accaatatct gtatactcag tgaaagtttt gtcttctagg taagttttca taatagtgac
     1801 ataccttatt ttttcatctg ggtgtctctc atcctgaaga atgggtgata tagttgaaat
     1861 aactcagttc gttttgtggt ttacgatttt tggcggatca caaaaatagg gctgagtccc
     1921 ttcaagtgag ttttgaaata tacaattata cccttcatcc tttaagttgt tgctttgttg
     1981 gctgttttca ttcacctcag tcacatagtt atctatgctc ctgaggattc gttcacttgc
     2041 cgtcgtgctg caactcgaaa tctattgggt atatatgtaa ttttcatatt ctgtttcatt
     2101 ttaacggtat tatttaaata tttatattta gaacattagg ttattagaat agataatttt
     2161 aggtaaatgt aattaattca accatcacgg agagtttatc agtcgaccga tgcccttgag
     2221 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
     2281 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat
     2341 tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt
     2401 cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca aacgtttcgg
     2461 cgagaagcag gccattatcg ccggcatggc ggccgacgcg ctgggctacg tcttgctggc
     2521 gttcgcgacg cgaggctgga tggccttccc cattatgatt cttctcgctt ccggcggcat
     2581 cgggatgccc gcgttgcagg ccatgctgtc caggcaggta gatgacgacc atcagggaca
     2641 gcttcaagga tcgctcgcgg ctcttaccag cctaacttcg atcattggac cgctgatcgt
     2701 cacggcgatt tatgccgcct cggcgagcac atggaacggg ttggcatgga ttgtaggcgc
     2761 cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc
     2821 gacctgaatg gaagccggcg gcacctcgct aacggattca ccactccaag aattggagcc
     2881 aatcaattct tgcggagaac tgtgaatgcg caaaccaacc cttggcagaa catatccatc
     2941 gcgtccgcca tctccagcag ccgcacgcgg cgcatctcgg gcagcgttgg gtcctggcca
     3001 cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt
     3061 actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa
     3121 acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg
     3181 gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc
     3241 tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg
     3301 atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt
     3361 aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt
     3421 atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa
     3481 aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa
     3541 ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct
     3601 gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     3661 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     3721 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     3781 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
     3841 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     3901 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3961 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4021 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4081 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4141 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4201 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4261 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4321 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4381 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4441 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4501 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     4561 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4621 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4681 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4741 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4801 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4861 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4921 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4981 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5041 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5101 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgctg
     5161 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5221 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5281 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5341 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5401 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5461 cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5521 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5581 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5641 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5701 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5761 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5821 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     5881 ggcgtatcac gaggcccttt cgtcttcaag aattctcatg tttgacagct tatcatcgat
     5941 aagctttaat gcggtagttt atcacagtta aattgctaac gcagtcaggc accgtgtatg
     6001 aaatctaaca atgcgctcat cgtcatcctc ggcaccgtca ccctggatgc tgtaggcata
     6061 ggcttggtta tgccggtact gccgggcctc ttgcgggata tcgtccattc cgacagcatc
     6121 gccagtcact atggcgtgct gctagcgcta tatgcgttga tgcaatttct atgcgcaccc
     6181 gttctcggag cactgtccga ccgctttggc cgccgcccag tcctgctcgc ttcgctactt
     6241 ggagccacta tcgactacgc gatcatggcg accacacccg tcctgtggat ccattttttt
     6301 aattaaataa aggaattaat attgatttga tagagcatgt aatcaataac ttctatgaaa
     6361 atgttgaaaa acaatacacg gaaacactta acttacaatt aaattgttaa aggagatgat
     6421 gataagaatg gtcagtcacc agataaaatg atctctggtc aattttagga aagctgaaaa
     6481 aacctgacca aatgcgtcgt aaattctatg cttagaaaga ctcggtgcca gtgtaatacc
     6541 taagatatgg tttttatttc catatctgtt cgaaaactta gaaataacaa gttcaacaat
     6601 aaccagggac tttaaatatg cctagatatg ctaattatca gataaacccc aaacagaata
     6661 ttaaaaattc acatgggaaa tcttcatcgt cagatttttc tagtgggtac ctttcattct
     6721 caaataattc gcttgatgac ccttttattc ggcagcaagt taagagagaa tttatttggg
     6781 agggacatat gaaggagatt gaagaagctt caagattagg tggctctggt ggtggcggtt
     6841 ctggcggcgg tggttctggc ggtgattata aggacgatga cgataaggat atggcaccga
     6901 aaaagaaacg caaagttggt attcatggtg ttccggcagc aatgggtggt agctatgcac
     6961 tgggtccgta tcagattagc gcaccgcagc tgcctgcata taatggtcag accgttggca
     7021 ccttctatta tgttaatgat gccggtggtc tggaaagcaa agtttttagc agcggtggtc
     7081 cgacaccgta tccgaattat gcaaatgcag gtcatgttga aggtcagagc gcactgttta
     7141 tgcgtgataa tggtattagc gaaggtctgg tgtttcataa taatccggaa ggcacctgtg
     7201 gtttttgcgt taatatgacc gaaacactgc tgcctgaaaa tgcaaaaatg accgttgttc
     7261 cgcctgaagg tagcggtaca ccgcatgaag ttggtgttta taccctgagt ggcacccctc
     7321 atgaagtggg cgtgtatacg ctgtttcagt gtcgtatttg tatgcgtaaa tttgcaacca
     7381 gcggtagtct gacccgtcat accaaaattc ataccggtga aaaaccgttt cagtgccgca
     7441 tctgcatgcg caattttagc cgtagcgatc atctgagcac ccatattcgt acccatacag
     7501 gcgaaaaacc ttttgcctgt gatatttgcg gtcgtaaatt cgccaccagt agcaatcgta
     7561 ccaaacacac aaaaattcac acccatccgc gtgcaccgat tccgaaacct tttcaatgta
     7621 gaatttgcat gcggaacttt tcccgttcag ataatctgag cgaacacatt cgcacacaca
     7681 caggtgagaa accatttgcg tgcgatatct gtggtagaaa atttgcctgg catagcagcc
     7741 tgcgtgttca tacaaaaatc catctgcgta gcggtggtag caccaatctg agtgatatca
     7801 ttgagaaaga aaccggtaaa cagctggtga ttcaagaaag cattctgatg ctgccagaag
     7861 aagtggaaga agttattggt aataaaccgg aaagcgatat tctggttcat accgcatatg
     7921 atgaaagcac cgatgaaaat gttatgctgc tgaccagtga tgcaccggaa tacaaaccgt
     7981 gggcattagt tattcaggat agcaatggtg agaacaaaat caaaatgctg taa
//
