LOCUS       sequence-444247-        6409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of the type I CBASS system encoded by Vibrio 
            campbellii.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6409)
  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 6409)
  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_Vibrio_campbellii
FEATURES             Location/Qualifiers
     source          1..6409
                     /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      4248..4319
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4380..4407
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4426..4517
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4518..4586,4587..5378)
                     /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(4736..4755)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5549..6137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6038..6057
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6291..6308
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(6323..6409,1..54)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6389..6408)
                     /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 atggcaaaag aagttactag
     1561 gtacataaaa gaaaaagtta aaactgcact gtggggaaag tcagcgggta ggtgtcaatt
     1621 taatggttgt aatagacctc tttaccaatc cccaatcaca ttggaacaag taaatatatc
     1681 agaagcagct cacatctatt cgttttccaa agatggcccg cgtgggtggg ggccgttagt
     1741 tacgaataaa gaggctctga acgatgttga taacttgatg cttatgtgtc acgactgtca
     1801 caaaaccata gatcaagata aaaaagggga aaagtactca gcagagcttc tactaaagtg
     1861 gaagaaagaa catgagcatc gtgtctggac agttgcaggt attaatcctt caaataagtc
     1921 tcatgtaatt ttttatggcg ctaacatagg cgaacaaatg tcccctttgc aataccaaga
     1981 agccgtaaat gctatgttcc cctctaagta cccctatagt gtagacgcca tagagctatc
     2041 aatgtccagt agtctaaaag acaaagatca agagttttgg attgctgaat caaagcattt
     2101 agaagaaata tttaaccagc aaataactcc taagttgagc ttaagcgact gtaatcattt
     2161 ctcactattt tctctagcgc caatgccttt actgataaag cttggttgct tgtttacgga
     2221 taaagttgac gttgaaactt atcaacccat cagagagcca aagacttggc ggtggcaatc
     2281 tccccccgat gattttgagt ttgatgtaac aatacctgat gtagtaaagc aatctcctgt
     2341 gttggtactt tcattgagtg cacatatcga tcacagtcga gtttatgatg tactgggtca
     2401 cgatgttgat atatgggagg ttagtactcc tatagacttt caacataacg attttattag
     2461 agcacctgaa cagctatcaa agtttagagt gacaatgaga aaactcttag aaaagatatc
     2521 tgctcaatat ggcttagatg ttcaactaaa agtctttcct gctatgcctg tttcatgttc
     2581 cgtagaactg gggcgtgttc ggatgccaaa agcaagtatg ccttgggaga tatttgacca
     2641 aagttattca actaaagtgt ttgaaccaac aataacaata cagggaggta agtgatgact
     2701 aatctcgtta caagagaaga agcccttttg gaggcagcaa ggaaactaga tattgccccc
     2761 cacaagtatc gtcaagctat tgagcgcttt aactcaatga aagctcacct agaacaagga
     2821 gtttatccag gaaccatgcg agtaccagaa gtgtacttgc aaggatcgtt caaactggga
     2881 acagaaattc gtccatttaa aaatagcaaa gatgcagact atgacattga tttagtctgc
     2941 aatctcgctc attcgaaaca gcacgtacaa ccaagtactg tgaaagagca agtcggtgat
     3001 aggatcagac aaaacggaac ttacgcaaga ttgctcgatg atgagggtaa gcgctgttgg
     3061 actttaaatt atgctgaaga agatggaatt ggcttccata tggatgtatt gccttcgata
     3121 tctgaaagcg gtctactggc gcacagttct gctattgctg cgacaaataa aaatgaacaa
     3181 gaacaatatt cttggacaac aagcaatccc aaaggcttcg cacaatggtt ttatgaaaaa
     3241 aatggggttg catttgaacg ggaaaaacta actcagaaaa cgcaaatctt taacaaccag
     3301 cggcagatgt acggttccgt caatgatgtg cctaacattc atgtaaaaac gcctttgcaa
     3361 agggcaattc aactacttaa gcgacatcgt gacgttaggt tttctcagtc tgaagtagaa
     3421 aacttcaagc ctatttccat gatcattagt gttcttgcgg caaagtgcta ccaaaatgag
     3481 acgacaattt ttgaaacttt acgaaatcta atttcgcggc tagagcagca agctaatcag
     3541 ttaagctccc agtttatata taaccagctt gatgcaagtt cactttactc attaattaca
     3601 cgcacgcagg atggaaagtg gataatttcc aatccaacaa atccaggaga gaactttgct
     3661 gatagatggc atgaagatga tcatgcaagg gccaaggcat ttttccaatg gattggatgg
     3721 cttcgagatg acttgataaa tatcaatcac ctagttgaaa gtgatttgtt taccaagtct
     3781 ttagttaatg tttatagacc aacgaacctt ccagctttaa attttcaggt ctcgcacaaa
     3841 gatacgggca gctggccgat agtattaaac ccttcttata gtgcatcggt taagggaagc
     3901 tataaaacga atgtattttg gaaagggttt aagtcgggtc aatcactact gaaaggtaaa
     3961 caacttaaat ttaagcttca aactgatgtt cctaggccgt ttgacgtata ttggcaagtt
     4021 gtcaatacgg ggcaggaagc aagagaagcg agacagctcc gaggtgaaat ctctgaacat
     4081 gtgaatacct caagttccga aatttttaga actgagtcaa cctcgtatat aggtgatcac
     4141 tgggttgaag gtttcattat caagaatggt gtatgtgttg caaaaacagg agaatttatt
     4201 gttagcgtga gagatacata tacaagatga taagaattcc caggcatcaa ataaaacgaa
     4261 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     4321 agctggtacc atatggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     4381 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct
     4441 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4501 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4561 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4621 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4681 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4741 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     4801 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4861 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4921 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4981 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5041 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5101 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5161 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5221 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5281 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5341 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5401 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5461 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5521 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5581 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5641 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     5701 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5761 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5821 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5881 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5941 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6001 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6061 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6121 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6181 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6241 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     6301 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     6361 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgc
//
