LOCUS       sequence-444240-        5469 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of the type I CBASS system encoded by Escherichia 
            coli RU1 BHI6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5469)
  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 5469)
  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_Escherichia_coli_RU1_BHI6
FEATURES             Location/Qualifiers
     source          1..5469
                     /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      3308..3379
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3440..3467
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3486..3577
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3578..3646,3647..4438)
                     /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(3796..3815)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4609..5197
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5098..5117
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5351..5368
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(5383..5469,1..54)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5449..5468)
                     /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 atgaaaagtc acgagtacat
     1561 tgtgatttgc ggttttaacc gcagtaaagt cggccgatat attgccattg ttgcctcagt
     1621 tatatcgttc gttctgattt tactagcact cacactaatc gaatggctga aaaactttca
     1681 cattaacagc cacatacctg cctcagtttt ttcacttatt agcgccggtg ggatctacat
     1741 ggggttgtat acatggtttg ataaaaatct ctggcataac acctacctcg ggaaattttt
     1801 atgtgtgcca aatcttgcag gccgctggaa tgttgaaggc catacccgat ctgaaggtgg
     1861 taaaccctgg gaaggtgagc tgagaattgt ccagtcatgg gacaaagtca gaattcatct
     1921 gaaaactcaa tcatcgcatt cggatagtgt gacagcctcc atcattcatg acgagggaat
     1981 tggttaccag ctgctttata actaccgcaa ccaacccaag acaggtgaag aacacctgac
     2041 ttcacatgtt ggattcgcag agttcaggtt cgacgacgga ctcaaatctg ctgaaggcca
     2101 ttattttaac ggtcaaggcc gggctactta cggaacaatg acaatcacta ggatcgacaa
     2161 tgtctaattc atttagtgcg cgaattgaac gcatgaagtc tcgtcgtaaa gggacattcg
     2221 atcaattaaa cgtagccaga gagtcaatca gtaaccagag gattgatgga ctggaaaact
     2281 atgctttgct agaaggtttc ctggatttaa atgaaagttg ggaaaccagg ggaaaacaag
     2341 attctgcaac ccgctatgtt ataggtgcca tgcaaccggt tgataaccgc tataccgaaa
     2401 tttcttttga aactgccaag cgtattgaga accaactggt aaagaaatta gatcttaatc
     2461 tggagtttcg tgtccaaggc tctgtaccac tggatattca tattaaaagt ttcagtgatg
     2521 tggatctgct gattatagac actcagatgc ttatttatga tagcgacggt attggccgct
     2581 acacccctac gaacaaaaat gatggggatg tcatccttga actcagggat gccgctaggg
     2641 atgccctgaa agcaactttc cctgcggcag atgttgatga caacaacgcc aaatctttga
     2701 gaataacggg agggtctttg cagagagaag tggatgtggt gccaagcatc tggtgggata
     2761 ccaaagagta ccaacatact aaagatgtag atcaacgtgg ggtcactatt atagataaaa
     2821 acacacgtca acgtatctac aacctacctt tcctccacat caaacgaatt aaagacaaat
     2881 gcgatcaatg taatggagga ttgcgtaagt ctattcgttt tcttaagaca ctaaaagcgg
     2941 atagtgaagc tgagggtact aaaattgaac tcagtagtta tgatattgcc tcgctgatgt
     3001 accacgcaga tggtaacaat ctccgacata gccaatacta cgagctggct gtccttgtag
     3061 agactcacag atggctaaat tatcttgcac agaaccctaa tgctgcaatg ctgctctacg
     3121 ttccaaatgg caccagaaag ataattgaca agaatgaaac ttttgccgaa ttgcttaaac
     3181 tgacgggaat ggtcaatagt attgtcactg aggtgctgcg tgaaataacc ggccagccta
     3241 ctgaatatta cataaacaga acaatggcaa aggcgttgcg ttcaccgtag taagaattcc
     3301 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     3361 tttgtcggtg aacgctctcc agctggtacc atatggcggg caggacgccc gccataaact
     3421 gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa
     3481 actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3541 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3601 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3661 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3721 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3781 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     3841 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3901 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3961 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4021 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4081 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4141 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4201 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4261 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4321 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4381 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4441 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4501 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4561 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4621 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4681 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4741 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4801 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4861 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4921 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4981 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5041 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5101 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5161 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5221 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5281 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     5341 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     5401 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
     5461 tgatgccgc
//
