LOCUS       sequence-444238-        5651 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of the type I CBASS system encoded by Escherichia 
            coli SLK172.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5651)
  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 5651)
  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_SLK172
FEATURES             Location/Qualifiers
     source          1..5651
                     /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      3490..3561
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3622..3649
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3668..3759
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3760..3828,3829..4620)
                     /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(3978..3997)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4791..5379
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5280..5299
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5533..5550
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(5565..5651,1..54)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5631..5650)
                     /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 atggcgacca cagtaatagc
     1561 tgcctttaac gaatttatga aagataccgt gaatatcaaa aaggcagata ccgatgacgc
     1621 gcgtgcaagt cgcgactggc ttatcggtaa gatgaatgat tttgagaagg acgataaatt
     1681 tccggtgagt tttccagcga tccatattgc ctttggctcc tttgccagaa gaaccaaaat
     1741 ccgtccgctt gatgatatcg atctgatgtt tggcttaacc gggcaaggtg ccacctatac
     1801 catcctgagt gaccggataa ctgtaacctc ctccggagag gggtcacgtc tgcacagtta
     1861 ccgccattcc ggagcagaca ccgtctgctc cgtcagaatt ctgaatgcct ttaaaaaccg
     1921 cctgcaggac attgcccagt acgcccaagc tgatatcagg cgtaaccagg aagctgtcac
     1981 gctgaaactg gtcagcaagg actggaattt tgacatcgtc ccctgcttta ttaccagcga
     2041 agatgccttt ggacggactt actacctgat ccccgatggc aacggccact ggaagtttac
     2101 cgacccccgt aaagacaggg acagagtcac tactatcaat gtgcagaata acggcaatgt
     2161 gctcaatgtc atcagagcag tcaaatactg gcagcggcga ccgacgatgc catccatgag
     2221 ttcctacctg ctggaaacgc tgattctgga ctattacgct ggcaggacat cctgctcatc
     2281 gtttgttgac atggagctgg aagcgctttt ccgccacctt ggtcaatccg tgcggtactc
     2341 tgtcaacgat cccaaaggca tacagggtga tatcaactca ctttcagccg aggcacgtaa
     2401 ggcgatatcg gacagatgtt atctcgatgc gcaaaaggta tcggaggcga gatggtttga
     2461 aaataataaa gagtatgaaa aatccatcaa taaatggcgc gatgtgttcg ggccgttctt
     2521 tcctgtttac gggtgaacca tgaatgccat ttacacaaac cagtcgacca gagaaaacct
     2581 ggatttactg tatgcgcagg cccgtgtcta cgacagggta aaaaactgga acagactgaa
     2641 ttttctcttc agcatcatcg taccgctcct gctttctttc gttaccgtat acaaccggag
     2701 ccgggaattt gtcgattcag agctgttgtc ttcgctgttg gggttatatg gactcctggt
     2761 gcttaccttt aacattgtta tcagtggaca catctcggct ctgcgcagga aggctgcctc
     2821 aatccaggaa atgtacgact gccgggtact tggcatccgg cgtaatgaac tgaaggtgga
     2881 ggagatccca cgggatgaca tcatccggga agctgcatat tttagagaca gcccggaaaa
     2941 agcacgaaag cgatttggcg aagaggggtg gtacgtcagt aaggtgtatg acgcgccgca
     3001 ggccgtaatg gcgctccttt gccacggaaa aaatctcggc tgggacaaat cgctcaggga
     3061 ggttctgcac gtattttatc tttccgcttt tattgtctcg cctgtcgcaa tgcttgttta
     3121 cggcatcgtc atgaaatccg ggctgaatga gatcctcttc tatatggtgt tcacactgcc
     3181 ggttatccgc tatttcttgc ttcagtttct ggataatcgc agcagtatga aaagaagcga
     3241 gaagctgaaa aagtacgttg agaaggaatt atccggcata agggtgtccg gaagggcaga
     3301 agaagagcaa ctgggttata cgctccggaa tatccaagat gagatgttta cctaccgagc
     3361 ctcatgtccg cccgttccca acggtattca gctgataatg aagcccaaaa atgaacgaat
     3421 ttatgttgat tactttgaaa caaatctgaa ggagctgcac cttcagaagt aataagaatt
     3481 cccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     3541 tgtttgtcgg tgaacgctct ccagctggta ccatatggcg ggcaggacgc ccgccataaa
     3601 ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac
     3661 aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3721 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3781 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     3841 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3901 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     3961 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     4021 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4081 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4141 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4201 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4261 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4321 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4381 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4441 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4501 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4561 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4621 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4681 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4741 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4801 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4861 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4921 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     4981 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5041 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5101 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5161 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5221 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5281 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5341 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5401 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5461 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     5521 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     5581 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc
     5641 tctgatgccg c
//
