LOCUS       sequence-388197-        6289 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains only regulatory genes; for generating empty PVCs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6289)
  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 6289)
  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-PVCpnf18-21
FEATURES             Location/Qualifiers
     source          1..6289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             516..707
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(701..723)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    809..949
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     864..883
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(964..981)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1135..1723)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1215..1234)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1894..2685,2686..2754))
                     /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     2517..2536
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2755..2859)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2927..2945
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(2972..2977,2978..2994,2995..3000)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 tctccagcag ccgcacgcgg cgcatctcgg gcagcgttgg gtcctggcca cgggtgcgca
       61 tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc
      121 agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga
      181 cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga
      241 agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct
      301 gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg atttttctct
      361 ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt aaccgggcat
      421 gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt atcattaccc
      481 ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa aaaaccgccc
      541 ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc
      601 tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt
      661 accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
      721 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
      781 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
      841 gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
      901 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc
      961 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     1021 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     1081 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1141 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1201 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1261 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1321 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1381 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1441 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1501 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1561 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1621 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1681 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1741 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     1801 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     1861 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     1921 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     1981 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     2041 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     2101 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     2161 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt
     2221 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     2281 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     2341 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     2401 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     2461 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa
     2521 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     2581 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     2641 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     2701 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     2761 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     2821 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     2881 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     2941 gaggcccttt cgtcttcaag aattctcatg tttgacagct tatcatcgat aagctttaat
     3001 gcggtagttt atcacagtta aattgctaac gcagtcaggc accgtgtatg aaatctaaca
     3061 atgcgctcat cgtcatcctc ggcaccgtca ccctggatgc tgtaggcata ggcttggtta
     3121 tgccggtact gccgggcctc ttgcgggata tcgtccattc cgacagcatc gccagtcact
     3181 atggcgtgct gctagcgcta tatgcgttga tgcaatttct atgcgcaccc gttctcggag
     3241 cactgtccga ccgctttggc cgccgcccag tcctgctcgc ttcgctactt ggagccacta
     3301 tcgactacgc gatcatggcg accacacccg tcctgtggat cctatggatt ttcatgtcat
     3361 caggagaata ttcaaaattc tgtgtcacac aaaatatttt acagcgtaat cacaccatta
     3421 agaggggcat caaaataaat ggttaactgg gacaatgttg aatttcattt ttataaggca
     3481 ttgtccgtat tatccaccat ccagatcaac ggattcagat ggtgtaaaga aagcatatag
     3541 ctggtaggta aaattaagaa gtgtagtttt ataaagtatt aatggagaac gatgaattta
     3601 aagtaaatgg aggaaggtta gtttcactat caaattacag gaccaaaaat atggagtatt
     3661 ccatcttctt tggtgaatta agtcttgaag acttcaacaa atattgcctg gaactatggg
     3721 cctgtgaatg gtattggagt atcatttata acgactattt cgataggaaa tttaaagtct
     3781 tgttgaaaat gactaatcgt ctaaaagtat gtctggagaa ataggtttaa tctctatatt
     3841 taacggtttt attttcatct atttactaat ggtaaatttt tttactcttt gaatatattg
     3901 gtgagtttgt tatccccatt agttaccagg gtattattaa tgagtgaaat tcaattttat
     3961 tgatattaaa gtcaaattta caaatgatga ctttattatt ttttatttat taattttaaa
     4021 ttattttttg ttaatcttta ctataaatga attttttgct tgcatggact tgattttttt
     4081 atatataaca ttgacaaata ttaattcata tatatgattg ttatagtgag ttatttaatt
     4141 gattcgacta acaatttcat tgtaattttt caagatagca gtcaattgct ataacaagca
     4201 tctttctcgt caaaggaaaa tgtgtatggc ctataagcag caacttgaaa gatgaaggat
     4261 ataatgatga ttaacttaac ggagagtaga tattagaagt agtagaagga caatttaaga
     4321 ctttataccc attaaaacaa aggggtaaaa taatgatgtt tgatgaaaat ttagaatgtg
     4381 ttaatcacga aattatcaat gaaaaattag atacgaaaat tcaccttgct aatccttatg
     4441 ccttttgggg gggaagggta aatgagaata taaatgggtt aattaaacaa cattctttaa
     4501 gaggaactga ctttaataaa atttttaacc ggaagataaa tttttttgta aatggaataa
     4561 ataattgccc ccgaaaaact cggaatataa aaccccctaa ttaattattt aaaggaataa
     4621 gagtttattt gctcgtagag taattatgtt ccattaatat gctaaattgc tcagtaaagt
     4681 tattaacttt ttttcaacgc cagtgaataa gtctgtttta ttaaataagg agttcaggtg
     4741 tttatttcaa aagaattatc aagctttatt gctgtagcta aaaataaatc aataaataaa
     4801 gcaacaacgg aactttttat tacacagtca ccgatttgcc gaagcttaaa aaaactggaa
     4861 ttttcattag gtgtaaaact ctttgtaaga aaaagccagg gattggcttt aacagaagaa
     4921 ggagaatgct tatataataa agttttgtca atttgtgaac ggcttcaaga aatagaaggt
     4981 gactacaaga atgaaaaatc aacatacaat gatctatatc aaaaaaatat atgaggttct
     5041 aatataataa gcggttctag ataactaaag tcatctatta ccaatatctg tatactcagt
     5101 gaaagttttg tcttctaggt aagttttcat aatagtgaca taccttattt tttcatctgg
     5161 gtgtctctca tcctgaagaa tgggtgatat agttgaaata actcagttcg ttttgtggtt
     5221 tacgattttt ggcggatcac aaaaataggg ctgagtccct tcaagtgagt tttgaaatat
     5281 acaattatac ccttcatcct ttaagttgtt gctttgttgg ctgttttcat tcacctcagt
     5341 cacatagtta tctatgctcc tgaggattcg ttcacttgcc gtcgtgctgc aactcgaaat
     5401 ctattgggta tatatgtaat tttcatattc tgtttcattt taacggtatt atttaaatat
     5461 ttatatttag aacattaggt tattagaata gataatttta ggtaaatgta attaattcaa
     5521 ccatcacgga gagtttatca gtcgaccgat gcccttgaga gccttcaacc cagtcagctc
     5581 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
     5641 gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg
     5701 ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc
     5761 tcaagccttc gtcactggtc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc
     5821 cggcatggcg gccgacgcgc tgggctacgt cttgctggcg ttcgcgacgc gaggctggat
     5881 ggccttcccc attatgattc ttctcgcttc cggcggcatc gggatgcccg cgttgcaggc
     5941 catgctgtcc aggcaggtag atgacgacca tcagggacag cttcaaggat cgctcgcggc
     6001 tcttaccagc ctaacttcga tcattggacc gctgatcgtc acggcgattt atgccgcctc
     6061 ggcgagcaca tggaacgggt tggcatggat tgtaggcgcc gccctatacc ttgtctgcct
     6121 ccccgcgttg cgtcgcggtg catggagccg ggccacctcg acctgaatgg aagccggcgg
     6181 cacctcgcta acggattcac cactccaaga attggagcca atcaattctt gcggagaact
     6241 gtgaatgcgc aaaccaaccc ttggcagaac atatccatcg cgtccgcca
//
