LOCUS       sequence-444239-        6369 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of the type I CBASS system encoded by 
            Stenotrophomonas maltophilia.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6369)
  AUTHORS   Richmond-Buccola D, Hobbs SJ, Garcia JM, Toyoda H, Gao J, Shao S, 
            Lee ASY, Kranzusch PJ
  TITLE     A large-scale type I CBASS antiphage screen identifies the phage 
            prohead protease as a key determinant of immune activation and 
            evasion.
  JOURNAL   Cell Host Microbe. 2024 Jul 10;32(7):1074-1088.e5. doi: 
            10.1016/j.chom.2024.05.021. Epub 2024 Jun 24.
  PUBMED    38917809
REFERENCE   2  (bases 1 to 6369)
  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: 
            "10.1016/j.chom.2024.05.021"; journalName: "Cell Host Microbe"; 
            date: "2024-07-10- 10"; volume: "32"; issue: "7"; pages: 
            "1074-1088.e5"
COMMENT     Sequence Label: pBAD02_CBASS_Stenotrophomonas_maltophilia
FEATURES             Location/Qualifiers
     source          1..6369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     140..162
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(318..1196)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1223..1507
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1430..1449
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     terminator      4208..4279
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4340..4367
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4386..4477
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4478..4546,4547..5338)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4696..4715)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5509..6097
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5998..6017
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6251..6268
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(6283..6369,1..54)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6349..6368)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
       61 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
      121 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
      181 aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc
      241 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt
      301 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa
      361 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag
      421 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc
      481 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct
      541 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg
      601 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt
      661 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac
      721 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg
      781 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag
      841 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga
      901 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga
      961 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga
     1021 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc
     1081 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc
     1141 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt
     1201 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt
     1261 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc
     1321 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat
     1381 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca
     1441 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt
     1501 tctccatacc cgttttttgg gctaacagga ggaattaacc atggtcagtg ctgtagagcc
     1561 gcgttggcat ggcgatagct accagtcccg ttttttctgg acccacgccg cgttgctgct
     1621 ggatccccag caaccccacg tagttgaggt tgccttcgag gcggatggcc ccaaggcctt
     1681 tgacgatgtc gtcgtccggt acaaccctgg tcgctcacgg cgctccgggc cgggctacgt
     1741 caccgccgac tatcatcaga ttaaatggca tgccaacaaa gccggccggt tcgggtttgc
     1801 cgacctgacc aaccccaagt tcatcaatgc cgccaaacac tcgattctgg agcggctgcg
     1861 cgatgcccaa aaggtggccg gcgaaggtgc agcgtttcac ttggtgacca ccgatcgcat
     1921 cgtggatgga gatcccttgg gcaagttagt gggcggccac gacagttcgt tacgactgca
     1981 ggtgctcaag gaaggaaaga ccgacaacag cgaaatggga agggtacgca aatgctggcg
     2041 cgagcatcta ggcttgagca gcgatgagga gctctatgcg atcttggacc ggttccacat
     2101 ccaagatagt cagcctgact tggaaaaaat gcgtcgtgag gtcagtaccc agtttcgtct
     2161 ggtgggcttg gccggtcatg agaacgaaac ggtatttctc tacgatgatg ccgccaaaca
     2221 gctggtaagt aagaagatca attgcttcga ccgagctggc tttcatcagt ggtgcacgga
     2281 cgaaaagtgg tatctggttc gcaagcctaa accgcgccga agtgtggcca tcgcaaccta
     2341 tagcccacgg ccgaccccgc tgcacatggc agacgccgag ccggacaaca ccctgaactt
     2401 ggctgaccga tttaccggtc ggtggctaaa agatggacac agctgggacg aggtacaggg
     2461 gcaggtacgc ctcttcctca atgccaaact ggcgcaggcc ccttccatgc gcctgttctt
     2521 ggagacgcat tcatccattg cctatctcgc cggagcggcg ctacgtttca aggagggcgc
     2581 cgatgttgag gttgttcagc gcggtgacaa aaaccccggg atcgtgtggg atgcccaaga
     2641 cggtaccgca gggccggagc cgctgattga ggtgcagaca ctgggggatg ggggcgacat
     2701 cgcggtcgct gtaggtctga gccagaacgt gggccagaag gtggggcact tcgtcgcgga
     2761 tcacctacct caggtcggca tgctgatcgg cgtatcgctg gttgacggtg ctggccccat
     2821 gaccattgcc ggcgggcagc atgctgcggc aattgctcag cacattgccg agttggttaa
     2881 agatacccgc aagccgggca acacgacgca ccttttcctc gccaccccca acgcttttgc
     2941 tttcctgctg ggccaacaca ccgacgcgat cggccgctgc attccttatg agtacgacat
     3001 ggctggccag atcgacggaa gctaccaccc cacctttacc acttaagggc atcgacatgg
     3061 aacttcgtcc ccagtttgct gaatttctgc aggacatccg ccccagcgag agaaaccgcg
     3121 aggaatggaa gcgtggctcg aatacgcttc gcacccgcct gatggctgac gccgagttgt
     3181 cgccgatgat tgcggccacc ttcctgcagg gtagcgtacg ccggtctacg gccgtacgac
     3241 cgaccggcgg caagcgctcg gatgtggatg tggtgatcgt cacgaccatt gactacaacc
     3301 gggaaactcc acgggtggcg atgaagcgat ttgaaccctt cttggagcgg cactaccaag
     3361 ggaagtgggc ccagcaggat cgttcgttcg ggatcgaaat gtcctacgtc gacctggacc
     3421 tagtggtgac tgcgctgccg gcagagccgg ccactcgcgc cgcactggaa tcgttgtatc
     3481 gcagccaggc agtacagttg gatgagtccc tggaagagcg cagcgactgg gtgctaaaca
     3541 agggatggga tccgcagcgt gcgaacggag gcttggtcag cctcactgaa gaagcgtcta
     3601 cctggcgcga tgagccgctg atgctgcctg accgtccggt gcagaaatgg ggacctaccc
     3661 atccgctggc gcagatccag tgggcggcag ctaagaaccg caactgcaat ggtcatttca
     3721 tcaacgtggt gcggtccatc aagtggtggc gacatgagca tgccgacacg ctgcccaagt
     3781 accccaaagg ctatccgctc gagcacatga tcggtgcggt cttgcccgat cacatccaga
     3841 gtgtggcgga aggagtcacc ttggcgctgg aggggattcg tgacacttgg gccgcctatg
     3901 ctgcggcggg cgtcgttcca tttcttgccg atcacggcgt gccctcccat aacgtcttgc
     3961 aacgtctggg ggcggaagat ttccgcgcct tcatggccaa ggcaggtgaa gcggccgatc
     4021 ttgcccgtcg cgcactgaac gagaccgaca aggatgttag cggcgcactg tggcgagagc
     4081 tgctgggcaa tagcttcccc ccgcctggcc ccggtggcgg tgatcgtggg cgcgggttcg
     4141 ccgcaccggc cgcccctgcc gtgccgcggc agtctgggcg gtttgcctga taagaattcc
     4201 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     4261 tttgtcggtg aacgctctcc agctggtacc atatggcggg caggacgccc gccataaact
     4321 gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa
     4381 actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4441 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4501 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4561 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4621 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4681 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     4741 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4801 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4861 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4921 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4981 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5041 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5101 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5161 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5221 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5281 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5341 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5401 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5461 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5521 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5581 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5641 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5701 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5761 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5821 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5881 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5941 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6001 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6061 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6121 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6181 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     6241 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     6301 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
     6361 tgatgccgc
//
