LOCUS       sequence-388196-        9141 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Loads HiBiT-tagged Pdp1 into PVCs; for payload loading assays; also 
            contains Pnf.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9141)
  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 9141)
  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-HiBiT
FEATURES             Location/Qualifiers
     source          1..9141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             6389..6580
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6574..6596)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6682..6822
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6737..6756
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6837..6854)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7008..7596)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7088..7107)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7767..8558,8559..8627))
                     /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     8390..8409
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8628..8732)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8800..8818
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(8845..8850,8851..8867,8868..8873)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 agtcctgctc gcttcgctac ttggagccac tatcgactac gcgatcatgg cgaccacacc
       61 cgtcctgtgg atccattttt ttaattaaat aaaggaatta atattgattt gatagagcat
      121 gtaatcaata acttctatga aaatgttgaa aaacaataca cggaaacact taacttacaa
      181 ttaaattgtt aaaggagatg atgataagaa tggtcagtca ccagataaaa tgatctctgg
      241 tcaattttag gaaagctgaa aaaacctgac caaatgcgtc gtaaattcta tgcttagaaa
      301 gactcggtgc cagtgtaata cctaagatat ggtttttatt tccatatctg ttcgaaaact
      361 tagaaataac aagttcaaca ataaccaggg actttaaata tgcctagata tgctaattat
      421 cagataaacc ccaaacagaa tattaaaaat tcacatggga aatcttcatc gtcagatttt
      481 tctagtgggt acctttcatt ctcaaataat tcgcttgatg acccttttat tcggcagcaa
      541 gttaagagag aatttatttg ggagggacat atgaaggaga ttgaagaagc ttcaagatta
      601 ggtaattttg ctgtcagttt tcgggctgcg ggcgggccaa cactcagagc tttgggcaaa
      661 ggtgcggcag ccaaggggca tgatattttg gaaaaaacca ttaaacccgg ctcaattaac
      721 aaagcttatc ccaaagacga agcttcagat gtaataaaga aagtacagga agcgggtatt
      781 gaaggctatg taggccattg ggataaaaaa actggccgcc ttttgggtat ttacatgagt
      841 agtggtcacg gtttatcaga tgaacaggtc aatggaaaaa tttatcctat agacttaaac
      901 aatctggaag cgagtctctc agccttgaaa gcaaaagaga actgggctgc actacctttt
      961 actggtgact atgatatgca tgatatgatt agtttcacag gtcaacctca ttcagtacca
     1021 tccaactcat cagaggaaag gaaaattata gatcgcatta accgattggt tgctcgatct
     1081 gattcaaacc ggccatttgg agatatagag cataatgtta tcaggcatgg agcccaggta
     1141 agctacccgg cttttgctat ggataaagaa aaagaggaaa taaaaaaaca cggggggatt
     1201 gtcaaggctg ttgctgaacc cggtgaattt cctgtcgcta ttgttagcaa aggaaaatgg
     1261 acaattgcca acaacattga tgaactaaat cagttttata atagtatcgg tgcaaaaatg
     1321 aaagtgagct ggaagccggg tgctgaaaat cctggattcg tttctaatcc acaaagaccg
     1381 ggcatggcca gatttagccg gaaaagaggg agctccggtg tgagcggctg gcggctgttc
     1441 aagaagatta gctgatatgg attttcatgt catcaggaga atattcaaaa ttctgtgtca
     1501 cacaaaatat tttacagcgt aatcacacca ttaagagggg catcaaaata aatggttaac
     1561 tgggacaatg ttgaatttca tttttataag gcattgtccg tattatccac catccagatc
     1621 aacggattca gatggtgtaa agaaagcata tagctggtag gtaaaattaa gaagtgtagt
     1681 tttataaagt attaatggag aacgatgaat ttaaagtaaa tggaggaagg ttagtttcac
     1741 tatcaaatta caggaccaaa aatatggagt attccatctt ctttggtgaa ttaagtcttg
     1801 aagacttcaa caaatattgc ctggaactat gggcctgtga atggtattgg agtatcattt
     1861 ataacgacta tttcgatagg aaatttaaag tcttgttgaa aatgactaat cgtctaaaag
     1921 tatgtctgga gaaataggtt taatctctat atttaacggt tttattttca tctatttact
     1981 aatggtaaat ttttttactc tttgaatata ttggtgagtt tgttatcccc attagttacc
     2041 agggtattat taatgagtga aattcaattt tattgatatt aaagtcaaat ttacaaatga
     2101 tgactttatt attttttatt tattaatttt aaattatttt ttgttaatct ttactataaa
     2161 tgaatttttt gcttgcatgg acttgatttt tttatatata acattgacaa atattaattc
     2221 atatatatga ttgttatagt gagttattta attgattcga ctaacaattt cattgtaatt
     2281 tttcaagata gcagtcaatt gctataacaa gcatctttct cgtcaaagga aaatgtgtat
     2341 ggcctataag cagcaacttg aaagatgaag gatataatga tgattaactt aacggagagt
     2401 agatattaga agtagtagaa ggacaattta agactttata cccattaaaa caaaggggta
     2461 aaataatgat gtttgatgaa aatttagaat gtgttaatca cgaaattatc aatgaaaaat
     2521 tagatacgaa aattcacctt gctaatcctt atgccttttg ggggggaagg gtaaatgaga
     2581 atataaatgg gttaattaaa caacattctt taagaggaac tgactttaat aaaattttta
     2641 accggaagat aaattttttt gtaaatggaa taaataattg cccccgaaaa actcggaata
     2701 taaaaccccc taattaatta tttaaaggaa taagagttta tttgctcgta gagtaattat
     2761 gttccattaa tatgctaaat tgctcagtaa agttattaac tttttttcaa cgccagtgaa
     2821 taagtctgtt ttattaaata aggagttcag gtgtttattt caaaagaatt atcaagcttt
     2881 attgctgtag ctaaaaataa atcaataaat aaagcaacaa cggaactttt tattacacag
     2941 tcaccgattt gccgaagctt aaaaaaactg gaattttcat taggtgtaaa actctttgta
     3001 agaaaaagcc agggattggc tttaacagaa gaaggagaat gcttatataa taaagttttg
     3061 tcaatttgtg aacggcttca agaaatagaa ggtgactaca agaatgaaaa atcaacatac
     3121 aatgatctat atcaaaaaaa tatatgaggt tctaatataa taagcggttc tagataacta
     3181 aagtcatcta ttaccaatat ctgtatactc agtgaaagtt ttgtcttcta ggtaagtttt
     3241 cataatagtg acatacctta ttttttcatc tgggtgtctc tcatcctgaa gaatgggtga
     3301 tatagttgaa ataactcagt tcgttttgtg gtttacgatt tttggcggat cacaaaaata
     3361 gggctgagtc ccttcaagtg agttttgaaa tatacaatta tacccttcat cctttaagtt
     3421 gttgctttgt tggctgtttt cattcacctc agtcacatag ttatctatgc tcctgaggat
     3481 tcgttcactt gccgtcgtgc tgcaactcga aatctattgg gtatatatgt aattttcata
     3541 ttctgtttca ttttaacggt attatttaaa tatttatatt tagaacatta ggttattaga
     3601 atagataatt ttaggtaaat gtaattaatt caaccatcac ggagagttta tcaatgttaa
     3661 aatatgctaa tcctcagacc gtagccacac aacgtactaa aaatactgcg aagaaaccgc
     3721 catcatcaac ctcttttgat gggcaccttg aactttcaaa tggtgaaaac cagccttacg
     3781 aaggccataa gattaggaaa atcaaaggat tgagacaaca tcttgcggat aggagtctta
     3841 ataaagggca tatttccccg ttgatgaata agggattatt agtaggtagt aaggatgtgt
     3901 caattgacat cccagttata gctcatcgtt atgacagcag ccatcaactt acagatgcgg
     3961 agcctttgaa agctgatagt cactccaatc atctagaccc tttttatggt gttattgcag
     4021 gatttcgtgg tgatcaggtg acatcatcag agtcgggttc aggatcaata ggcgtacatt
     4081 gggggaaaaa tacgctggat tccaatatta tgggtgttaa tgtggttaat ggtgcatcgg
     4141 gaaccgtggg aataagaatt gctttgaagg atattcagca tgggtccccg gttattgtca
     4201 cgtctggagc attgagcggt tgtactatgg tatattcggt gaaaaatggc tatttttttg
     4261 cctatcatac ggggcagaaa ccgggtaata atgaatggaa aacaggcagg cagggtgttg
     4321 ttgctaccta tctgagccat caagcccttt cgccggatag tgaacctatg acggttggtg
     4381 aacagaataa cgatttagtg aatatttttg caaattatga tcagagtgtt attacctata
     4441 tgggaaaacc gggagtgctg attgataaaa tggcagaaaa tgttggtgtt tttaattatg
     4501 acgaaattaa gcctgaaaaa cctgctatta gagctggcta cagttatgcg ttgctggcta
     4561 aggacgataa aggaaaggtg aacgttaagg tactatcgga agatgttatc gtttcatcag
     4621 gtaagcaggg aaatactgtt aaggccatta attcacttaa aaaacggttg ttataatttt
     4681 gcccaatgag ggcaggaggt acggattttc attagcactt atcgaagcgg atatatgttt
     4741 gtgactacat tgtcagaaaa tttcattcat caacttgaat tagaacagtg tccatataac
     4801 ttatttagaa taaaggaagg aaattaatct tagatacaat gataagggaa ataaaataga
     4861 aagtagagtt aattgtatta ataaggtgct ggagctttac ttacaaaata tcaatgattt
     4921 ttggtttcgt ataagtaatt aatttagagt aatgaaaaat tatttgtggg ttaaatttta
     4981 tttattgaaa attcattggt ttattttatg aaataggaag gtaatagcgg tattattcgg
     5041 ttgaaataac cgtatagata aatagtagga tttagataaa ttatttcata tatgacaatt
     5101 tttactatta tatggggttt gtaagtcgac cgatgccctt gagagccttc aacccagtca
     5161 gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta
     5221 tcatgcaact cgtaggacag gtgccggcag cgctctgggt cattttcggc gaggaccgct
     5281 ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt attcggaatc ttgcacgccc
     5341 tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag caggccatta
     5401 tcgccggcat ggcggccgac gcgctgggct acgtcttgct ggcgttcgcg acgcgaggct
     5461 ggatggcctt ccccattatg attcttctcg cttccggcgg catcgggatg cccgcgttgc
     5521 aggccatgct gtccaggcag gtagatgacg accatcaggg acagcttcaa ggatcgctcg
     5581 cggctcttac cagcctaact tcgatcattg gaccgctgat cgtcacggcg atttatgccg
     5641 cctcggcgag cacatggaac gggttggcat ggattgtagg cgccgcccta taccttgtct
     5701 gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga atggaagccg
     5761 gcggcacctc gctaacggat tcaccactcc aagaattgga gccaatcaat tcttgcggag
     5821 aactgtgaat gcgcaaacca acccttggca gaacatatcc atcgcgtccg ccatctccag
     5881 cagccgcacg cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcgt
     5941 gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga
     6001 atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc
     6061 aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc
     6121 gccctgcacc attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac
     6181 acctacatct gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg
     6241 ccgcatccat accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc
     6301 atcagtaacc cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa
     6361 cagaaatccc ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat
     6421 ggcccgcttt atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc
     6481 ggatgaacag gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag
     6541 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     6601 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     6661 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
     6721 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     6781 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg
     6841 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6901 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6961 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     7021 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     7081 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     7141 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     7201 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     7261 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     7321 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7381 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7441 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7501 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7561 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7621 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7681 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7741 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7801 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7861 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7921 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7981 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     8041 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca
     8101 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     8161 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     8221 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     8281 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     8341 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg
     8401 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8461 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8521 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8581 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8641 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8701 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8761 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     8821 tttcgtcttc aagaattctc atgtttgaca gcttatcatc gataagcttt aatgcggtag
     8881 tttatcacag ttaaattgct aacgcagtca ggcaccgtgt atgaaatcta acaatgcgct
     8941 catcgtcatc ctcggcaccg tcaccctgga tgctgtaggc ataggcttgg ttatgccggt
     9001 actgccgggc ctcttgcggg atatcgtcca ttccgacagc atcgccagtc actatggcgt
     9061 gctgctagcg ctatatgcgt tgatgcaatt tctatgcgca cccgttctcg gagcactgtc
     9121 cgaccgcttt ggccgccgcc c
//
