LOCUS       sequence-444241-        7397 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of the type I CBASS system encoded by Escherichia 
            coli K71-77.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7397)
  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 7397)
  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_K71-77
FEATURES             Location/Qualifiers
     source          1..7397
                     /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      5236..5307
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5368..5395
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5414..5505
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5506..5574,5575..6366)
                     /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(5724..5743)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6537..7125
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7026..7045
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7279..7296
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(7311..7397,1..54)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7377..7396)
                     /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 atgacttact ctgtttcgcc
     1561 gtcatcacta ctcaccgagt acggcaacga caacatatgc cgtgttctcg cacttgatgg
     1621 tggcggtgca aagggctttt atacgctggg tgtcctcaag gaaattgagg ctatgcttgg
     1681 gtgcccatta tataagaggt ttgatcttgt atttggtact agcaccggtg cgattattgc
     1741 cgcgctcatc gctctaggat acgaagtaga tcaaatacac gcactgtaca ccgaacatgt
     1801 accacgagtt atgtccagcc ggtcggctgc ggcaagaaca atggcgctcc aggatttggc
     1861 taaggaagtt tttcaagaca agacattcga agatgttctg atgggtattg gcatcgtcgc
     1921 aactcgatgg atgaccgaaa gaccaatgat cttcaaaggt aatgttgtgc aggcccatgg
     1981 tcgcaaggga accttcagtc ctggattcgg ggtcagcatc gctgatgcag tgcaagcgtc
     2041 ttgctcggct tatccgttct tcgaaaggaa agtgattgtt accgcagcag gagataaggt
     2101 tgagttgatc gatggaggtt actgcgcaaa caaccccacc ttgtttgcaa tcgccgacgc
     2161 cactgtcgct ctgaaaaaag atcacaagga tatccgggtt ataaatgtcg gtgttggtat
     2221 ctatccagaa cctaaaccag gtctccttat gcggattgct aagaaatggc tcgcagtcca
     2281 gctgcttcaa aaaactttag agatcaatac acaatcgatg gatcaactac gcgacattct
     2341 attcaaagat attccgacga ttcggatcag cgacacattc gaacggcctg agatggcgac
     2401 agatttactt gagtacaatt tagataaact gaataccctt cgacagcgtg gtagggagtc
     2461 ttttggagct cgtgaggcac aattaagaga gtttctgata tgagtgaggc gaaataatat
     2521 gggaatacct gaatcacagc tcgatacatg gtcccaccaa ggatctattg cacagtctgc
     2581 ctcgacttat agcattataa agaatgcatt agagagcgca aacactaagt atcatggaaa
     2641 gaattttaaa gtattccttc agggctccta tggaaacgat actaacattt atgctgaaag
     2701 tgacgttgat gtagtcatat gtcttgatga tgtctactac agtgacctca cacagttatc
     2761 accagaagac aaagatgcgt atgaccgtgc atttgttcct gcaacctact cgtatactca
     2821 atttaagcaa gatgtgcttg aggctcttac agagcgcttc gggtctgatg ttaaggtcgg
     2881 agataaggcc atagttgtag cggcaaatgg aagtaggcgc aaagctgacg ttatcgcatc
     2941 aatgcagttt cgtcgttatt ggaaattcaa ggggcattac gactcacaat acgatgaggg
     3001 catctgtttc tttaatggcg ctggtgaacg gattgccaat taccccaagc agcattcaga
     3061 aaatctcacc ttaaaacatc aggccagtaa taaatggcta aagcccatgg ttcgcgtact
     3121 gaagaacctt cgaagtaaac tcattgctga cggaaaattg aagtcaggac ttgcaccttc
     3181 ctattacctt gaaggtctac tctacaacgt gccaaatgaa aagtttggca ccagttatgc
     3241 tgattgtttt gtcaatgcca tgaattggat tcagacagaa gcagataaag acaagctggt
     3301 atgcgccaat gaacagtatt acttgctttg ggaggggaca catacctcat gggagaaagc
     3361 cgatgcggaa gcgtttatcg acgctgcaat aaaaatgtgg aatgaatggt gaagtgagtc
     3421 gacaaaactg ctggtcttag ggcacggcaa acagcaagca ggagaaatcg aggtggcagg
     3481 tagcatcatt aaccttttta tgtggggata tcaggacagc tacagaattc acatccagat
     3541 tctggcacgt aatgtactga aaaaattggg tgcaccggct gatgcagaag tgctacttgt
     3601 tggtgctcgc cgccctggca gtaaaaatgc gaatccgata tgcgtagagc cagaagatgg
     3661 taagtggcat ctatctttgt ttgaaggatt gctggattca gttgaatcga tttacaagaa
     3721 tcatcggtta cagagtatga tttttggtga tgggccaagc atgcgcgaca agccagagtg
     3781 gatacgtcgc gactcagtgc gaacatcggt tagtaaggca cttgatgctt ttgataccga
     3841 gcaaaacgta acgtcatttt gcggagagtc taggcgaata gacgactatt atgtcacccc
     3901 agtcatacag attccaaacg acactttcga gaaattccct cctttgctct ccaagtctgt
     3961 tgacgaaagc caacaaggac atggatttcg tagtctgata catgcagcga tgtatgcggt
     4021 tcttcgtgag gctaccgagg aattgcataa ccctgatcca ggaagattta tccatggcag
     4081 tatgagagat gccgaggaaa taatacggat tgctgcaaac gactttctgc acaccgcagg
     4141 cctctcgatc gaacgaagat acattcatac tgatcttttc gatgctttga accttgtctc
     4201 ctctcttatg tacgaggggg cgaaaggcat aggtcagctc attctcgtcg atccagataa
     4261 cgaggcggtt gagttcctgg cgaagtttgt tgagccagtt ccctttcgtg agccacgatg
     4321 ggttcggaaa gtcttgcaga tggcggcatc aggtgtcggc attatcgcaa atagtcagta
     4381 catctatgga cttgggagac tgaagaagtc gcacgatccc agtgcacaag atgcgttcac
     4441 ggtagatttt atagatcatt atcactggca gttgtactgc ggtaaccaag cactgcttct
     4501 cagtcaatat gctgtgccca aacttccaca ggagcctttt gacaaagctg ccttcctggc
     4561 aaattacgcg agactatttc cgttgtcttc acaagatagt ggattgcact tgtggaacct
     4621 tctgatgacc caaaccagtc aggggcatgg gagcatgatt gtcgttgcgg aagatgcggc
     4681 ctgcgaggcg aatcgacttt gcaagcaagg tactagaatc gtgccaacta agctcactga
     4741 atcactcctt cgaagtgtca gcggtattga tggcacaata ctgcttgatc ctgctggtct
     4801 ttgccatgct atcgggatta ttctcgacgg tgaagcgacc gatgagtgta cgccatcaag
     4861 gggatctcgg tataattccg gagtgcgtta cgtacaaacg agcggcactc gaaggcttgc
     4921 gatcgtcgtg tcggatgatc gaacagtcga tatcattcct caaatcaagc gactgtgcag
     4981 tcgttcaaaa atagaacaat gtgtcgtaac actggaggcg gcaacgctcg acaactatca
     5041 tgattctcgg aattggttaa acgaccatcg cttctatatc aacgctgagc aatgcgctcg
     5101 aattaatgcc gtactcgaca aactagatgc cgttccaaaa gaagtaggtc ttatctattt
     5161 tgaaactgag agatttgaag ttgacccaga gatggacgaa agctatctca tagactgata
     5221 agaattccca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     5281 atctgttgtt tgtcggtgaa cgctctccag ctggtaccat atggcgggca ggacgcccgc
     5341 cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt
     5401 ttctacaaac tcttttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5461 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5521 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5581 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5641 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5701 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     5761 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5821 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5881 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5941 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6001 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6061 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6121 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6181 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6241 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6301 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6361 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6421 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6481 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6541 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6601 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6661 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     6721 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6781 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6841 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6901 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6961 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7021 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7081 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7141 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     7201 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     7261 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     7321 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     7381 atctgctctg atgccgc
//
