LOCUS       sequence-444236-        5002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of the type I CBASS system encoded by Escherichia 
            coli BWH59.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5002)
  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 5002)
  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_BWH59
FEATURES             Location/Qualifiers
     source          1..5002
                     /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      2841..2912
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2973..3000
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3019..3110
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3111..3179,3180..3971)
                     /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(3329..3348)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4142..4730
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4631..4650
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4884..4901
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(4916..5002,1..54)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4982..5001)
                     /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 atgaatcaga agctcatcca
     1561 aacagtcatt tctgagtgtc atcgtataga agaggacgct ctgtactctt ctaaatctca
     1621 ttataatgtt tctgatagat gggataagct gaatctgtgg ttcggtgtac cattagcaat
     1681 cttaaccgct ttgagtggta ttgccgcaat caagagtaca gccaatgctg ttgtagctat
     1741 ttccataact gctacaattc taactgcatt aaatacgtct cttaacccaa gtaagcgtgc
     1801 tgcacttcat aaatcagcag caaatgaata taataaatta aagaatgaag ttagaatatt
     1861 tagggaaata ttgacttcta attttgattt tactgataaa actttgattg aaaaacttac
     1921 gagttactct gatcgacgca atcaacttaa ttcaagttcg ccaaatattc ctcggtgggc
     1981 ttatgagaaa actcaaagag atgttaacca aggctttact aagtataata ttgacaagga
     2041 ataaacatga cagtcagtac ctatcttgag tctattaaaa gcaatgctta tcaaagagat
     2101 attacaataa cgaattcaat tgcaaccata aaaaacagac tgaatggtta ttttgctggt
     2161 gaacttgtta gtcattttgt atttggctca tattcacgta atacaatgct tcctcgtagc
     2221 tatgatcctt cttctgatgt tgattatatg gttgtattca aaaatgtaat atatcagcct
     2281 cagacctacc taaacaagct aagggatttt gtaaattatt actatcgtac atctgaaata
     2341 aagcaaagcc accctactat tcaattgaac cttaatcata ttactttcga gcttgtccct
     2401 gcatctcata atattttatt tggttatcaa atcccttcca accaggattt tttgaaccgt
     2461 tggatgaata ccgatcccaa tgcgttcaat actgatttaa ctaattgcaa taaaaataat
     2521 aacaatttaa ttaagcctct tattcgcata atgaaatatt ggaatgccaa tgcaggttat
     2581 gtctttgagt cgtatcaact tgaaaaagat attgttagct ttaactattt ttggtgctct
     2641 acccttaaag aatattttta ctctgctgta gaacaacttt caggttccta ttatctcgca
     2701 caatggaaaa aagataagat tcagcatcta aaaaattcag ttatgatgtc aaaatattat
     2761 gagcagttaa acaatcattt tgccgccgaa aatgaaataa agaaaatttt acccataaaa
     2821 taataagaat tcccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     2881 gttttatctg ttgtttgtcg gtgaacgctc tccagctggt accatatggc gggcaggacg
     2941 cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt
     3001 tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3061 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3121 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3181 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3241 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3301 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     3361 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3421 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3481 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3541 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3601 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3661 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3721 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3781 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3841 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3901 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3961 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4021 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4081 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4141 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4201 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4261 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     4321 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4381 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4441 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4501 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4561 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4621 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4681 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4741 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4801 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4861 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     4921 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
     4981 gtacaatctg ctctgatgcc gc
//
