LOCUS       sequence-444246-        5668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of the type I CBASS system encoded by Escherichia 
            coli CVM N37067PS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5668)
  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 5668)
  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_CVM_N37067PS
FEATURES             Location/Qualifiers
     source          1..5668
                     /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      3507..3578
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3639..3666
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3685..3776
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3777..3845,3846..4637)
                     /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(3995..4014)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4808..5396
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5297..5316
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5550..5567
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(5582..5668,1..54)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5648..5667)
                     /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 atgcaaacag cagagatgtt
     1561 tcgaaatttt ttgaacaatt tgaaaatttc agacgaacgt atgcaggcaa ttgcaacaca
     1621 ttataaaaat gccacaagaa gacttaacat cgagtttcgg ggggtagcga atggattcaa
     1681 caatcgattg aaagtcggtt cagcaggtcg ccgaacagcg gttagcaaaa catcagactt
     1741 ggatatgctg tatattatgc ccgcgagatt gtggggagat taccaatccg gtgttaatcc
     1801 acagcgccgt ttgcttcgcg atgtaaaaga agcattaaaa ctatctttta cccagcaaga
     1861 gatcaaggtt gatagattag ttgtacaaat catttttgac tcttttcata tagaggtgca
     1921 accagttttt gagactgatg aagggcattt taaatatcct gatactaagg ccaacgatgg
     1981 taatggaggg tggcgagtaa ctaaacctcg gatggaaatt accgaaatga gaaatttccg
     2041 taataacaaa agtagtaatc ttcataatct ttgccgcatg ttaagagcct ggaaaaatag
     2101 aaacaccgtt gatatggggg ggcttctcat tgacacattg gcttggagat ttctgaaaca
     2161 aacagatgat tatgataata ccggaatggt cagttatggg cttatgtgtc gagacttctt
     2221 tgactttttg catcaagaag atgtccatga acattatgca gcgataggaa gtgtgcagag
     2281 agttaaggtc tatagaaatt ttcagcggca agcaaaacga gcctttaaat tatgcacaaa
     2341 agcgattgaa gcttatgaga atggctcgac aaagaaatgc cacgaacact ggcgagaagt
     2401 ttttgggcta actttcccta aagcagttgt agaaatgcgt gactctcagg ggttagagtc
     2461 agctaatttc atcaatgagt cctatacggc tcagacgtgg cgcaatacgg aagagtttac
     2521 agatgaaaag tttgatgatg ttgatataag ataccccctc gaagttaact gcattgtcaa
     2581 agaaaacggg tatagggacc gtgcaattag ggcttacctg tcggatatgg ctcgttattg
     2641 gttaccaccg aatagaactt tatcattttc gatcaaccaa gcaagtttgg atatcattcc
     2701 tgagccttat gatatttact ggaaagtact gaatagaggt ttagaagctg agcagcgtga
     2761 tgaagttcga ggtcaaatag ttttaggtca agtaactcac actgaacata caaaatttag
     2821 aggtgatcac aaggtttggt gttacatagt taagaaccat atagtaattg cccaaaacgt
     2881 tatcgacata ccaatcgagt aattatcatg caaaaaaatg acttattatc tcagattgcc
     2941 tctgccggat ataatgttgg atttggcgct aagaaaaatt ttgctacatt tgatattgtt
     3001 agcaaactcc ctggctggat tggtattata tcactagcag tctctatttt aggcctttat
     3061 atggatgaaa tgagttctaa gaatattgca gccattttta tcattttaag cattggcagt
     3121 ttgtacatca atttctatga tggtgataaa aataaatatg aagaggctgg aaagaagcaa
     3181 acagcaatct ttagaaagat tgaaagattg tattacgatg caaaatcaga aagcgaaccg
     3241 gattctgtta aatatattgg gcaacttgac attctgttaa aagaatttga tgaaaccact
     3301 atcaccaaac aaatttgtgg aagcaattgg tatgctcatt ataaattctt tagtgaaatg
     3361 gaaaaaaggt ggcttgaaga tcagttgtca ttgactttct ggaaggataa agtccctaac
     3421 actttgaaat ttctcattct gatagttttc tgcgtcgttt tgctttcgat aatttatagg
     3481 tgtggttgat aagaattccc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     3541 cctttcgttt tatctgttgt ttgtcggtga acgctctcca gctggtacca tatggcgggc
     3601 aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg
     3661 cctttttgcg tttctacaaa ctcttttgtt tatttttcta aatacattca aatatgtatc
     3721 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3781 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3841 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3901 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3961 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     4021 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4081 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4141 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4201 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4261 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4321 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4381 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4441 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4501 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4561 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4621 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4681 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4741 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4801 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4861 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4921 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     4981 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5041 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5101 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5161 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5221 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5281 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5341 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5401 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5461 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5521 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     5581 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
     5641 ctctcagtac aatctgctct gatgccgc
//
