LOCUS       sequence-444245-        5631 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of the type I CBASS system encoded by Escherichia 
            coli BIDMC103.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5631)
  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 5631)
  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_BIDMC103
FEATURES             Location/Qualifiers
     source          1..5631
                     /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"
     CDS             complement(2850..2861)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     terminator      3470..3541
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3602..3629
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3648..3739
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3740..3808,3809..4600)
                     /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(3958..3977)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4771..5359
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5260..5279
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5513..5530
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(5545..5631,1..54)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5611..5630)
                     /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 atgagccgca gggaattgtt
     1561 cagcgaattt cttgaaaatt tgaatcttga ccttaaacag gcaaaaaaga tcagctatca
     1621 ctatcgaaaa ataaccaagt cattaaatct tgcgttcaga ggaacgagtt cccgggtagc
     1681 aaaccgcttg aaagtggggt cagtaggcag acatactgca ataaaaggca tatccgatct
     1741 ggacatgctt tatattatgc cccccaatca gtatgaatat tataaccgta aggataatgg
     1801 tcaggctgca ttgcttacag atgtcagaaa cattttagcg gaagaatatc ctgaccagac
     1861 ggtgaaaaag gacagactgg ttgttcagat catctttaaa aacttttatg tcgaggttca
     1921 acctgtattc agacaggatg atggcagctt taaattccct gaaagctata atgggggagc
     1981 atggcgcatt acaaaacctc ttcacgaaaa ggcggctatg accgcatttt cgcgagataa
     2041 atccaataat ctgcgcaagc tgtgtaaaat gatcagagcc tggaagaacc tgcatggcgt
     2101 caatatgggg ggcttactga tagatacact ggcgtatcga tttctttcat caacatcaga
     2161 ttatgacaat accggaaatg gcagcctggg agccctcgcc agagactttt ttgaatatct
     2221 gtcaaatgag gagaaaaggg aaaggtatct cgcgttaggc agtaatcagt atgtacgtgt
     2281 taaatctccc tggtttggcc gggcagcgaa acatgcgtat gaattatgtt gtgatgcctt
     2341 agacgctgag gggtctgcca gtgaaaatga cctctggcgc gaagtgttcg gacgagcctt
     2401 ccccagacgt aaggtgggta ttatggaagc cagccttggt ctggaatcgc atgcggcaga
     2461 tgcagctccc tggatcgata ccgaagaatt tatagaggat aaatatccgg tggatatcag
     2521 gcactcactc aaccttgact gtaccgtaac ccaagatggt ttccctccca gaagcctgag
     2581 agagatgctt actagaggat ttcgcctctc cgctcggaaa tctctccttt tccgggcaga
     2641 cttgacagaa atggaggcag aggaacccta caccgttatg tggaaggtac tgaacgtggg
     2701 tgacgaagca cgcagaagaa atatgatccg gggacaaatc gtttcggacg gcggctactg
     2761 tacaaagaaa gagaccaccg acttccgcgg agatcacgtg gtagaatgct atgtgataaa
     2821 aaatgaggtt gtggttgccc gggcccagat tgaagttccc ataaactgag gtttttatga
     2881 cgcaatcaga gatgctgaaa ttaattgcca gacatggtta taacgttgga tttggcgcta
     2941 agaaaaactt tgccacctat gatctaattt gtaaggcatc cggttggatt agctttattt
     3001 ctcttgctgt cggtgtgttc gctcttttta ttcctgagct agccacaaac gtaatttccg
     3061 cgatattaat tatatttggt gttgctacaa tgtacattca gttttatgac tcggaaaagc
     3121 agagttatga aagtgcgggg attgagcaga caaaaatttt taatgaactt gaagttattt
     3181 accgcaacgt aaaaagcgat gcaaactttg actatcagca tacacaacaa tccatcgacg
     3241 ctatcatgtc ccggttttac ggcacggtca tcagcaaaca gatcttcgga agtaactggt
     3301 acgcacatta taaattttac tgtgaatttg agaaaacatg ggttgaagaa gaactaaact
     3361 tgacactgaa ggacaaaatc ccgctttcgc tctgctttct cgccggtctg gcaatcattt
     3421 tcattatagc tgtcatctgg aaaatgctct gataagaatt cccaggcatc aaataaaacg
     3481 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     3541 ccagctggta ccatatggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag
     3601 cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt
     3661 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3721 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     3781 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3841 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3901 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3961 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     4021 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4081 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4141 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4201 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4261 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4321 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4381 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4441 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4501 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4561 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4621 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4681 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4741 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4801 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4861 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     4921 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4981 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5041 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5101 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5161 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5221 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5281 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5341 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5401 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5461 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     5521 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     5581 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg c
//
