LOCUS       sequence-444248-        7943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of the type I CBASS system encoded by Vibrio 
            vulnificus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7943)
  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 7943)
  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_vulnificus
FEATURES             Location/Qualifiers
     source          1..7943
                     /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      5782..5853
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5914..5941
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5960..6051
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6052..6120,6121..6912)
                     /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(6270..6289)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7083..7671
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7572..7591
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7825..7842
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(7857..7943,1..54)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7923..7942)
                     /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 atgactgatg agaggttaaa
     1561 ttatcatcaa gcatgttttt atcccataga gtcattagac aagctcacag agtttctatc
     1621 gggtattaag acttcttttg cgtgggagca tgttggatta cttactgatg aacatgtgaa
     1681 aaaccaggtc tatatattct attcacatga gttccgagcc agctgtcggc ttaacgtaac
     1741 tgatgcggag cttaaggatg ctctaatgtt gatgggctgt ggggacaatc ttgaagttcg
     1801 ccacctccag ggccgaaacc tttctcaaga cgaatatgga acaatagagt cgctatttaa
     1861 aatatgggat tcctgtgtta accaatctat agtcttgtca gtcgatgagg ttgtcgacta
     1921 tatttccagg aacatcgaac agcgatttaa taaaggtcgc ggcaaagatt tcacacagcc
     1981 gacaattaat caagttatgc gtgatagtca tggctattgt atgtttgaag gctgcgggga
     2041 taggttagat attgattatc tgacgggggc atctggcaac tattcatact tggctcacaa
     2101 tgttgcgtct tccgagagtg gcccaagagg tataccttat ctatccgaag aactttctaa
     2161 tgagccagct aatatcctac ttctatgcga caagcatcac cgattaatag atcgagttgc
     2221 gtgtgcggat taccctgcat ccagacttac tcgaatgcga tcagagtttt taattaatac
     2281 tgagagatta cttgaaggac tgagtttcca gcctattcct gttttttcag tactttggcc
     2341 ggttggtggg catgttgttt ctcctccaga gttaagagac attgcagaga gcttatctag
     2401 gttaagaact cgcattagag gtacaatgaa taaccttagt gataatgaac gccgatatag
     2461 aaccaaacct gaacggtttg ccaaagaaat ggatgaaatt gtcacagagg aagcagaaac
     2521 catagttaat caaacaaaga attatggata tcgagcggcg ctattcgcat ttgggccaat
     2581 gcctgctttg gtaggattag gggcgagcct aggaaataaa tctgaaatca cgccaatgct
     2641 cagatatcga gatgggaact gctgggtttg gcctcagaat tctcggtcag aaacacctta
     2701 caaggtgaac tggaagaacg aaataggcaa atcatctgaa gtcactatct ctatagcatt
     2761 aacccaatat cctgaagtaa tgaaaaagac tgcagcagcg ctcgggtatc ctgagattga
     2821 aatcgtagcc aatcaaatgg gtaatgcggc tctcccgcac ccggataacg ggttggaatt
     2881 aaaaagtgaa tttcacgcaa tactgcaaaa acttcgtgac caatttgatg ttaatcgtgt
     2941 tcatttgctt atctgtgcat ctaacgctgc ttgcgtattt gtaggccaag ccgtcgatct
     3001 ctatcagcct gcacttcttg tatatgactt cgatggtgac ataatgaggt ctaagctgtc
     3061 gataactcaa gtaaacggaa aaattaatct atcaatacct caatgagtag ccattgttag
     3121 aagagccact ctgtagcatt actaaaattt ggtcaattgg cgctttacag aggagtaagg
     3181 atacatatga cttttaagat tttaagcatt gatggcggtg gaatacgggg agtttaccct
     3241 gcgcacatac ttaaatgttt cgaagaaaaa ctcggaatca atctcttaga gtcattcgac
     3301 atgatagctg gcactagtac ggggtccata attgcggctg gtgttgcctg cgacatccga
     3361 gcaaccgaaa tagttaacat gtacaaagaa catggcgaag atattttcaa aaagaaaaag
     3421 tcgagaatac cttttaagaa gtttcgaagt attacgcaac cgttgcttga aagcgtctat
     3481 gacagtggca gcttatctaa agttcttaaa agtgttttca acgatagtac tcttgggcaa
     3541 attgccaaac ctctgattct acctgcaacg gatgtaggta acggcggagt tcatgtattt
     3601 aaatcggctt atgacccaac gtttaatcgt gataggttgg ttaaagtcag agatgctgtt
     3661 ctagcatctt gctcagcccc aacatatttt gaccctcata aagttgatga gtacttgcta
     3721 tcagatggtg gtctgtgggc aaacaaccca gcattagtgg ctgttatcga tgcccaaaaa
     3781 cgactgggga taaattacga tgatatacaa gtagtatcca tcggcaccgg tcattccaaa
     3841 gtcgcctatg gtttaaagca caagggagac tgggggcttg caacgggctg gaggcataaa
     3901 gaatttataa gtttcatgct atcgcttcag agccaaagcg cgttgaacta tctaaagctt
     3961 catttgaagc ctgagcaaat attaagaatt gattttgagt ccaatactga tctgccactc
     4021 gatgatgttt ctgaaatcga taatttgatc agtttagccg acaagaattt tacatatgga
     4081 agtaaagaga ttagcgattt tttcttagga ggaaatgtat agtgactcaa atagtttcta
     4141 gggaagaggc tctattaaca gcggcgaaaa agttggatat ttcaccgact aaatatagac
     4201 aagcgattga gcgctttaat tcaatgagaa cccacctaga gcaaggtaac tatcctggaa
     4261 catttcgagt cccagaggta taccttcaag gctcttttaa acttggaaca gaaatccgcc
     4321 catttaaaaa tagtaaagat gcagattatg atattgattt ggtctgcaac ttgggacatt
     4381 caaaacaggc agtacatgcc tccactgtaa aaaatcaggt tggggatcgg attaaagaga
     4441 atggcactta ttcccgattg ttagatgatg aaggaaagcg ctgctggact ctcaactatg
     4501 ctgaagatga cggtgttgga ttccatatgg atatactccc ttctgttgca gaagacagct
     4561 tattagcaca acatcaatca atttctgcta caaataaaga tcaacaaaat agatactctt
     4621 ggacaacgag taacccgaag ggatttgctg aatggtttta tgaaagaaat ggcactgcat
     4681 ttgaacaagc aaaattaact cagaagcaac gcatcttcaa caatcaaaga cagttgtaca
     4741 attcaattga agatgttcca aatatccacg ttaaaacgcc attgcagagg gctattcaac
     4801 tattaaagag acaccgtgac gtaaggtttt cacaatcaga tattgaaaca ttcaaaccaa
     4861 tctcaatgat cattacggta ttagctgcca acagttataa caatgaagca actatctatg
     4921 acacgttgag aaatctaata tcatctttaa accagcattc tcatcaatta agtaatagct
     4981 ttgtttttaa tgattactac gcaagaaata cctacacatt aataactcgt acgtctgatg
     5041 gaaagtggct tatacccaac cctgctaatc caggagaaaa cttcgctgac cgatggcatg
     5101 aggagaataa cgcaagagca aatgctttct tccaatgggt gggttggctc agagacgact
     5161 ttattaatat tgataactct atcgatgaag ataagtttac taagtcactt gttgatgtat
     5221 acaaaccaac aaatttacct gcactaagct ttaatgtgcc ccataaggaa cagcttcgat
     5281 acccaatcgt ttcacacatc tactcagcga atataaaagc aagttataaa tctaacatat
     5341 attggaaaga gtttcagtct ggacagcctt tactcaaagg aaaggagtta aagttcaaac
     5401 ttaagactaa tgttcctgca ccattcgacg tttattggca ggtagtaaac acaggtcaag
     5461 aggcggcaaa tgtagggcag cttagaggtg aaatatctcc tcaagggtac acgacgttag
     5521 gcgaaattta tcgaacagaa tctacctctt acaccggtga tcattgggta gagggattcg
     5581 tcatcaaaga tagtgtatgc gttgcgagaa ctggtgaatt tgtcgtaagc gtcagagata
     5641 ggttctctgt atgaataatg tgaacttcca agaagtatta agtgcaaaaa ttactgcact
     5701 tatgctgcta actaatctag gctttgtaat attaaaaact tggtcaactg atggtattct
     5761 gtgataagaa ttcccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     5821 cgttttatct gttgtttgtc ggtgaacgct ctccagctgg taccatatgg cgggcaggac
     5881 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
     5941 ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc
     6001 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     6061 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6121 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6181 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6241 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     6301 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6361 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6421 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6481 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6541 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6601 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6661 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6721 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6781 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6841 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6901 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6961 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     7021 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     7081 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     7141 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     7201 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     7261 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     7321 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     7381 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     7441 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7501 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7561 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7621 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7681 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7741 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     7801 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     7861 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     7921 agtacaatct gctctgatgc cgc
//
