LOCUS       sequence-487001-        8124 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PVC cargo/regulatory region - deleted toxin cargos (Pdp1/Pnf) - 
            vegfa sgRNA - Cre loaded into the PVC tube via Pdp1 (NTD) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8124)
  AUTHORS   Kreitz J, Yang V, Friedrich MJ, Pham J, Macrae RK, Zhang F
  TITLE     Targeted delivery of diverse biomolecules with engineered bacterial 
            nanosyringes.
  JOURNAL   Nat Biotechnol. 2025 Aug 12. doi: 10.1038/s41587-025-02774-x.
  PUBMED    40796978
REFERENCE   2  (bases 1 to 8124)
  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 
            Biotechnol. 2025 Aug 12. doi: 10.1038/s41587-025-02774-x."
COMMENT     Sequence Label: pBR322-sgRNA-pvc18-20-Pdp1_NTD-Cre
FEATURES             Location/Qualifiers
     source          1..8124
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             467..658
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(652..674)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    760..900
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     815..834
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(915..932)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1086..1674)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1166..1185)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1845..2636,2637..2705))
                     /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     2468..2487
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2706..2810)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2855..2889
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     protein_bind    2922..2940
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    2977..2995
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     promoter        join(3110..3115,3116..3132,3133..3138)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4052..4075
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4079..4099
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4100..5128
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLQ
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
ORIGIN
        1 acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct
       61 tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa
      121 aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct
      181 ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg atctgcatcg caggatgctg
      241 ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt
      301 gatttttctc tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag
      361 taaccgggca tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg
      421 tatcattacc cccatgaaca gaaatccccc ttacacggag gcatcagtga ccaaacagga
      481 aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa
      541 actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc
      601 tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
      661 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
      721 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
      781 tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga
      841 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
      901 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
      961 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     1021 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1081 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1141 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1201 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1261 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     1321 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     1381 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     1441 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     1501 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     1561 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     1621 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     1681 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     1741 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     1801 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     1861 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     1921 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     1981 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     2041 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     2101 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     2161 gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     2221 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     2281 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     2341 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     2401 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     2461 acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     2521 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     2581 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     2641 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     2701 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     2761 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     2821 cgaaaagtgc cacctgacgt ctaagaaacc cctcttgaca gctagctcag tcctaggtat
     2881 aatgctagcg gtgagtgagt gtgtgcgtgg tttgagagct agggccctga agaagggccc
     2941 tagcaagttc aaataaggct agtccgttat caacttgggc cctgaagaag ggcccaagtg
     3001 gcaccgagtc ggtgcttttt ttccgaaaaa aaaaccccgc ccctgacagg gcggggtttt
     3061 ttttaattaa cgtatcacga ggccctttcg tcttcaagaa ttctcatgtt tgacagctta
     3121 tcatcgataa gctttaatgc ggtagtttat cacagttaaa ttgctaacgc agtcaggcac
     3181 cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg caccgtcacc ctggatgctg
     3241 taggcatagg cttggttatg ccggtactgc cgggcctctt gcgggatatc gtccattccg
     3301 acagcatcgc cagtcactat ggcgtgctgc tagcgctata tgcgttgatg caatttctat
     3361 gcgcacccgt tctcggagca ctgtccgacc gctttggccg ccgcccagtc ctgctcgctt
     3421 cgctacttgg agccactatc gactacgcga tcatggcgac cacacccgtc ctgtggatcc
     3481 atttttttaa ttaaataaag gaattaatat tgatttgata gagcatgtaa tcaataactt
     3541 ctatgaaaat gttgaaaaac aatacacgga aacacttaac ttacaattaa attgttaaag
     3601 gagatgatga taagaatggt cagtcaccag ataaaatgat ctctggtcaa ttttaggaaa
     3661 gctgaaaaaa cctgaccaaa tgcgtcgtaa attctatgct tagaaagact cggtgccagt
     3721 gtaataccta agatatggtt tttatttcca tatctgttcg aaaacttaga aataacaagt
     3781 tcaacaataa ccagggactt taaatatgcc tagatatgct aattatcaga taaaccccaa
     3841 acagaatatt aaaaattcac atgggaaatc ttcatcgtca gatttttcta gtgggtacct
     3901 ttcattctca aataattcgc ttgatgaccc ttttattcgg cagcaagtta agagagaatt
     3961 tatttgggag ggacatatga aggagattga agaagcttca agattaggtg gctctggtgg
     4021 tggcggttct ggcggcggtg gttctggcgg tgattataag gacgatgacg ataaggatcc
     4081 caagaaaaag cgcaaggtat ccaatttact gaccgtacac caaaatttgc ctgcattacc
     4141 ggtcgatgca acgagtgatg aggttcgcaa gaacctgatg gacatgttca gggatcgcca
     4201 ggcgttttct gagcatacct ggaaaatgct tcagtccgtt tgccggtcgt gggcggcatg
     4261 gtgcaagttg aataaccgga aatggtttcc cgcagaacct gaagatgttc gcgattatct
     4321 tctatatctt caggcgcgcg gtctggcagt aaaaactatc cagcaacatt tgggccagct
     4381 aaacatgctt catcgtcggt ccgggctgcc acgaccaagt gacagcaatg ctgtttcact
     4441 ggttatgcgg cggatccgaa aagaaaacgt tgatgccggt gaacgtgcaa aacaggctct
     4501 agcgttcgaa cgcactgatt tcgaccaggt tcgttcactc atggaaaata gcgatcgctg
     4561 ccaggatata cgtaatctgg catttctggg gattgcttat aacaccctgt tacgtatagc
     4621 cgaaattgcc aggatcaggg ttaaagatat ctcacgtact gacggtggga gaatgttaat
     4681 ccatattggc agaacgaaaa cgctggttag caccgctggt gtagagaagg cacttagcct
     4741 gggggtaact aaactggtcg agcgatggat ttccgtctct ggtgtagctg atgatccgaa
     4801 taactacctg ttttgccggg tcagaaaaaa tggtgttgcc gcgccatctg ccaccagcca
     4861 gctatcaact cgcgccctgg aagggatttt tgaagcaact catcgattga tttacggcgc
     4921 taaggatgac tctggtcaga gatacctggc ctggtctgga cacagtgccc gtgtcggagc
     4981 cgcgcgagat atggcccgcg ctggagtttc aataccggag atcatgcaag ctggtggctg
     5041 gaccaatgta aatattgtca tgaactatat ccgtaacctg gatagtgaaa caggggcaat
     5101 ggtgcgcctg ctggaagatg gcgattaata tggattttca tgtcatcagg agaatattca
     5161 aaattctgtg tcacacaaaa tattttacag cgtaatcaca ccattaagag gggcatcaaa
     5221 ataaatggtt aactgggaca atgttgaatt tcatttttat aaggcattgt ccgtattatc
     5281 caccatccag atcaacggat tcagatggtg taaagaaagc atatagctgg taggtaaaat
     5341 taagaagtgt agttttataa agtattaatg gagaacgatg aatttaaagt aaatggagga
     5401 aggttagttt cactatcaaa ttacaggacc aaaaatatgg agtattccat cttctttggt
     5461 gaattaagtc ttgaagactt caacaaatat tgcctggaac tatgggcctg tgaatggtat
     5521 tggagtatca tttataacga ctatttcgat aggaaattta aagtcttgtt gaaaatgact
     5581 aatcgtctaa aagtatgtct ggagaaatag gtttaatctc tatatttaac ggttttattt
     5641 tcatctattt actaatggta aattttttta ctctttgaat atattggtga gtttgttatc
     5701 cccattagtt accagggtat tattaatgag tgaaattcaa ttttattgat attaaagtca
     5761 aatttacaaa tgatgacttt attatttttt atttattaat tttaaattat tttttgttaa
     5821 tctttactat aaatgaattt tttgcttgca tggacttgat ttttttatat ataacattga
     5881 caaatattaa ttcatatata tgattgttat agtgagttat ttaattgatt cgactaacaa
     5941 tttcattgta atttttcaag atagcagtca attgctataa caagcatctt tctcgtcaaa
     6001 ggaaaatgtg tatggcctat aagcagcaac ttgaaagatg aaggatataa tgatgattaa
     6061 cttaacggag agtagatatt agaagtagta gaaggacaat ttaagacttt atacccatta
     6121 aaacaaaggg gtaaaataat gatgtttgat gaaaatttag aatgtgttaa tcacgaaatt
     6181 atcaatgaaa aattagatac gaaaattcac cttgctaatc cttatgcctt ttggggggga
     6241 agggtaaatg agaatataaa tgggttaatt aaacaacatt ctttaagagg aactgacttt
     6301 aataaaattt ttaaccggaa gataaatttt tttgtaaatg gaataaataa ttgcccccga
     6361 aaaactcgga atataaaacc ccctaattaa ttatttaaag gaataagagt ttatttgctc
     6421 gtagagtaat tatgttccat taatatgcta aattgctcag taaagttatt aacttttttt
     6481 caacgccagt gaataagtct gttttattaa ataaggagtt caggtgttta tttcaaaaga
     6541 attatcaagc tttattgctg tagctaaaaa taaatcaata aataaagcaa caacggaact
     6601 ttttattaca cagtcaccga tttgccgaag cttaaaaaaa ctggaatttt cattaggtgt
     6661 aaaactcttt gtaagaaaaa gccagggatt ggctttaaca gaagaaggag aatgcttata
     6721 taataaagtt ttgtcaattt gtgaacggct tcaagaaata gaaggtgact acaagaatga
     6781 aaaatcaaca tacaatgatc tatatcaaaa aaatatatga ggttctaata taataatcgg
     6841 ttctagataa ctaaagtcat ctattaccaa tatctgtata ctcagtgaaa gttttgtctt
     6901 ctaggtaagt tttcataata gtgacatacc ttattttttc atctgggtgt ctctcatcct
     6961 gaagaatggg tgatatagtt gaaataactc agttcgtttt gtggtttacg atttttggcg
     7021 gatcacaaaa atagggctga gtcccttcaa gtgagttttg aaatatacaa ttataccctt
     7081 catcctttaa gttgttgctt tgttggctgt tttcattcac ctcagtcaca tagttatcta
     7141 tgctcctgag gattcgttca cttgccgtcg tgctgcaact cgaaatctat tgggtatata
     7201 tgtaattttc atattctgtt tcattttaac ggtattattt aaatatttat atttagaaca
     7261 ttaggttatt agaatagata attttaggta aatgtaatta attcaaccat cacggagagt
     7321 ttatcagtcg accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc
     7381 ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac
     7441 aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga
     7501 tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca
     7561 ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccg
     7621 acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc ttccccatta
     7681 tgattcttct cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc
     7741 aggtagatga cgaccatcag ggacagcttc aaggatcgct cgcggctctt accagcctaa
     7801 cttcgatcat tggaccgctg atcgtcacgg cgatttatgc cgcctcggcg agcacatgga
     7861 acgggttggc atggattgta ggcgccgccc tataccttgt ctgcctcccc gcgttgcgtc
     7921 gcggtgcatg gagccgggcc acctcgacct gaatggaagc cggcggcacc tcgctaacgg
     7981 attcaccact ccaagaattg gagccaatca attcttgcgg agaactgtga atgcgcaaac
     8041 caacccttgg cagaacatat ccatcgcgtc cgccatctcc agcagccgca cgcggcgcat
     8101 ctcgggcagc gttgggtcct ggcc
//
