LOCUS       sequence-388687-        7796 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For loading R half of zinc finger deaminase targeting human TRAC 
            locus (ZFD_TRAC-NC-Right-G1397-C_DddA); also contains PVC regulatory
            genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7796)
  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 7796)
  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_R
FEATURES             Location/Qualifiers
     source          1..7796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(165..753)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(245..264)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(924..1715,1716..1784))
                     /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     1547..1566
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1785..1889)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1957..1975
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(2002..2007,2008..2024,2025..2030)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             2944..2967
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2977..2997
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3625..3876
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7342..7533
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(7527..7549)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    7635..7775
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     7690..7709
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(join(7790..7796,1..11))
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
       61 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
      121 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
      181 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
      241 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
      301 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
      361 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
      421 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
      481 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
      541 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
      601 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
      661 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
      721 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
      781 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
      841 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
      901 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
      961 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     1021 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     1081 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     1141 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     1201 tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc
     1261 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     1321 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     1381 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     1441 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     1501 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca
     1561 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     1621 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     1681 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     1741 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     1801 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     1861 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     1921 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     1981 cgtcttcaag aattctcatg tttgacagct tatcatcgat aagctttaat gcggtagttt
     2041 atcacagtta aattgctaac gcagtcaggc accgtgtatg aaatctaaca atgcgctcat
     2101 cgtcatcctc ggcaccgtca ccctggatgc tgtaggcata ggcttggtta tgccggtact
     2161 gccgggcctc ttgcgggata tcgtccattc cgacagcatc gccagtcact atggcgtgct
     2221 gctagcgcta tatgcgttga tgcaatttct atgcgcaccc gttctcggag cactgtccga
     2281 ccgctttggc cgccgcccag tcctgctcgc ttcgctactt ggagccacta tcgactacgc
     2341 gatcatggcg accacacccg tcctgtggat ccattttttt aattaaataa aggaattaat
     2401 attgatttga tagagcatgt aatcaataac ttctatgaaa atgttgaaaa acaatacacg
     2461 gaaacactta acttacaatt aaattgttaa aggagatgat gataagaatg gtcagtcacc
     2521 agataaaatg atctctggtc aattttagga aagctgaaaa aacctgacca aatgcgtcgt
     2581 aaattctatg cttagaaaga ctcggtgcca gtgtaatacc taagatatgg tttttatttc
     2641 catatctgtt cgaaaactta gaaataacaa gttcaacaat aaccagggac tttaaatatg
     2701 cctagatatg ctaattatca gataaacccc aaacagaata ttaaaaattc acatgggaaa
     2761 tcttcatcgt cagatttttc tagtgggtac ctttcattct caaataattc gcttgatgac
     2821 ccttttattc ggcagcaagt taagagagaa tttatttggg agggacatat gaaggagatt
     2881 gaagaagctt caagattagg tggctctggt ggtggcggtt ctggcggcgg tggttctggc
     2941 ggtgattata aggacgatga cgataaggat atggcaccga aaaagaaacg caaagttggt
     3001 attcatggtg ttccggcagc aatggcagaa cgtccgtttc agtgtcgtat ttgtatgcgt
     3061 aattttagcc gtagcgatca tctgagcacc catattcgta cccataccgg tgaaaaaccg
     3121 tttgcatgtg atatttgcgg tcgtaaattt gcagatcgta gccatctggc acgtcatacc
     3181 aaaattcata ccggtagcca gaaacctttt cagtgccgca tctgcatgcg gaaatttgca
     3241 ctgaaacagc atctgaacga acacacaaaa atccacacag gcgaaaaacc attccaatgc
     3301 cggatttgca tgcgcaactt tagtcagagc ggtaacctgg ctcgtcatat tcgcacacat
     3361 actggcgaga agccttttgc ctgcgatatc tgtggtcgca aattcgcaca taatagcagc
     3421 ctgaaagatc acacgaaaat tcatctgcgt agcggtacac cgcatgaagt tggtgtttat
     3481 accctgagtg gcacccctca tgaagtgggc gtgtatacgc tggcaattcc ggttaaacgt
     3541 ggtgcaacag gtgaaaccaa agtttttacc ggcaatagca atagcccgaa aagcccgacc
     3601 aaaggtggtt gtagcggtgg tagcaccaat ctgagcgata tcattgagaa agaaaccggt
     3661 aaacagctgg tgattcaaga aagcattctg atgctgcctg aagaagtgga agaagttatt
     3721 ggtaataaac cggaaagcga tattctggtt cataccgcat atgatgaaag caccgatgaa
     3781 aatgttatgc tgctgaccag tgatgcaccg gaatacaaac cgtgggcatt agttattcag
     3841 gatagcaatg gtgagaacaa aatcaaaatg ctgtaataat atggattttc atgtcatcag
     3901 gagaatattc aaaattctgt gtcacacaaa atattttaca gcgtaatcac accattaaga
     3961 ggggcatcaa aataaatggt taactgggac aatgttgaat ttcattttta taaggcattg
     4021 tccgtattat ccaccatcca gatcaacgga ttcagatggt gtaaagaaag catatagctg
     4081 gtaggtaaaa ttaagaagtg tagttttata aagtattaat ggagaacgat gaatttaaag
     4141 taaatggagg aaggttagtt tcactatcaa attacaggac caaaaatatg gagtattcca
     4201 tcttctttgg tgaattaagt cttgaagact tcaacaaata ttgcctggaa ctatgggcct
     4261 gtgaatggta ttggagtatc atttataacg actatttcga taggaaattt aaagtcttgt
     4321 tgaaaatgac taatcgtcta aaagtatgtc tggagaaata ggtttaatct ctatatttaa
     4381 cggttttatt ttcatctatt tactaatggt aaattttttt actctttgaa tatattggtg
     4441 agtttgttat ccccattagt taccagggta ttattaatga gtgaaattca attttattga
     4501 tattaaagtc aaatttacaa atgatgactt tattattttt tatttattaa ttttaaatta
     4561 ttttttgtta atctttacta taaatgaatt ttttgcttgc atggacttga tttttttata
     4621 tataacattg acaaatatta attcatatat atgattgtta tagtgagtta tttaattgat
     4681 tcgactaaca atttcattgt aatttttcaa gatagcagtc aattgctata acaagcatct
     4741 ttctcgtcaa aggaaaatgt gtatggccta taagcagcaa cttgaaagat gaaggatata
     4801 atgatgatta acttaacgga gagtagatat tagaagtagt agaaggacaa tttaagactt
     4861 tatacccatt aaaacaaagg ggtaaaataa tgatgtttga tgaaaattta gaatgtgtta
     4921 atcacgaaat tatcaatgaa aaattagata cgaaaattca ccttgctaat ccttatgcct
     4981 tttggggggg aagggtaaat gagaatataa atgggttaat taaacaacat tctttaagag
     5041 gaactgactt taataaaatt tttaaccgga agataaattt ttttgtaaat ggaataaata
     5101 attgcccccg aaaaactcgg aatataaaac cccctaatta attatttaaa ggaataagag
     5161 tttatttgct cgtagagtaa ttatgttcca ttaatatgct aaattgctca gtaaagttat
     5221 taactttttt tcaacgccag tgaataagtc tgttttatta aataaggagt tcaggtgttt
     5281 atttcaaaag aattatcaag ctttattgct gtagctaaaa ataaatcaat aaataaagca
     5341 acaacggaac tttttattac acagtcaccg atttgccgaa gcttaaaaaa actggaattt
     5401 tcattaggtg taaaactctt tgtaagaaaa agccagggat tggctttaac agaagaagga
     5461 gaatgcttat ataataaagt tttgtcaatt tgtgaacggc ttcaagaaat agaaggtgac
     5521 tacaagaatg aaaaatcaac atacaatgat ctatatcaaa aaaatatatg aggttctaat
     5581 ataataagcg gttctagata actaaagtca tctattacca atatctgtat actcagtgaa
     5641 agttttgtct tctaggtaag ttttcataat agtgacatac cttatttttt catctgggtg
     5701 tctctcatcc tgaagaatgg gtgatatagt tgaaataact cagttcgttt tgtggtttac
     5761 gatttttggc ggatcacaaa aatagggctg agtcccttca agtgagtttt gaaatataca
     5821 attataccct tcatccttta agttgttgct ttgttggctg ttttcattca cctcagtcac
     5881 atagttatct atgctcctga ggattcgttc acttgccgtc gtgctgcaac tcgaaatcta
     5941 ttgggtatat atgtaatttt catattctgt ttcattttaa cggtattatt taaatattta
     6001 tatttagaac attaggttat tagaatagat aattttaggt aaatgtaatt aattcaacca
     6061 tcacggagag tttatcagtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt
     6121 ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca
     6181 actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc gctttcgctg
     6241 gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg ccctcgctca
     6301 agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca ttatcgccgg
     6361 catggcggcc gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag gctggatggc
     6421 cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt tgcaggccat
     6481 gctgtccagg caggtagatg acgaccatca gggacagctt caaggatcgc tcgcggctct
     6541 taccagccta acttcgatca ttggaccgct gatcgtcacg gcgatttatg ccgcctcggc
     6601 gagcacatgg aacgggttgg catggattgt aggcgccgcc ctataccttg tctgcctccc
     6661 cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc tgaatggaag ccggcggcac
     6721 ctcgctaacg gattcaccac tccaagaatt ggagccaatc aattcttgcg gagaactgtg
     6781 aatgcgcaaa ccaacccttg gcagaacata tccatcgcgt ccgccatctc cagcagccgc
     6841 acgcggcgca tctcgggcag cgttgggtcc tggccacggg tgcgcatgat cgtgctcctg
     6901 tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg
     6961 atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca
     7021 tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc
     7081 accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca
     7141 tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc
     7201 cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta
     7261 acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat
     7321 cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc
     7381 tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa
     7441 caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc
     7501 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     7561 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     7621 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
     7681 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     7741 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctc
//
