LOCUS       sequence-444237-        5639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of the type I CBASS system encoded by Escherichia 
            coli 703128.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5639)
  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 5639)
  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_703128
FEATURES             Location/Qualifiers
     source          1..5639
                     /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      3478..3549
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3610..3637
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3656..3747
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3748..3816,3817..4608)
                     /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(3966..3985)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4779..5367
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5268..5287
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5521..5538
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(5553..5639,1..54)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5619..5638)
                     /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 atggctctta cggtgactgc
     1561 tgcgtttaac gagtttcagc gcaatattgt aaatctagat agctctcaga ctgatcgagc
     1621 cagagccagt agagattggt tattgggcag aatgaacact tttccaaata atgatgttta
     1681 tttccccata atataccctg atatacatac aggttttggt tcttttgcta gacgcacaaa
     1741 aatccgccct ttagatgata tagatcttat gtttggtctc gatggtgatg attgtgtata
     1801 cagcgagagt gatggaaaaa taataatcac tgcgcgtgaa ggaaccgttc gcttaaaata
     1861 ctataagcat gatggcacaa attttattaa ctctagaaaa ataataaaca gttttatctc
     1921 aagcctaaat caaattccgc agtatgataa ggctgatgta aaaagaaatc aggaagcagc
     1981 tactctgaag ctaaaatcct atgactggaa ttttgatata gttcctgcat tcattactac
     2041 tccagattca tttggcaaga catattattt aataccagat gggaatggtc attggaagaa
     2101 gacagatcca cgaattgaca agcaaaaagt tactgatctt aatgttaagc tctcaggaaa
     2161 catgttaaat gtcattcgtg taataaaata ttggcaaaaa cgtccgacca tgccttcaat
     2221 gagtagttac ttgttagaga caatgctact taactacttc aacggtcgat cagaatgtta
     2281 tcaatgggtc gatttagaga ttgtcaatgt tttaggttac ttagcctctg ctatatttag
     2341 tccagtgtac gatcacaaag gcatacaagg aaacataaac aaccttagtg atgatgataa
     2401 attaaagata tctattcgct gttatcttga ccaaggaaaa gcaacggaag cccgtaatta
     2461 cgaaagccaa aataaccaca gagcgtccat taacaaatgg caggatgttt ttggtgtcaa
     2521 cttcccatca tacggatgaa aaatgagtca aatatttttg cgccaaaatg atagtaaaaa
     2581 cattgagtta ctaatggcaa tgaggcggac ttactcacag gttaaatttg ttaactgggt
     2641 agtgtttttt cttagtgtcg taatgtctgt cgctattgta atatttacta cagtgataaa
     2701 gaaaatgggg ctattccctc atttgactat agctccttat ctcggttatt acggtataat
     2761 cgtgttggtc atcacaatta ttttttccac aatgatttcc tcaatgaaaa aatgtgcagc
     2821 cactattcag gaaatgtttg actgtgatgt cttacacatt ccatggagtg aattaaaagt
     2881 agggaagccg gttggcagag aaaatgtctt taaggcttca aggtattaca aaaaaagaaa
     2941 caaaaatgat gaatttatca actggtatct aaataaagat tacgatgcta atgaaaacgt
     3001 attggcatta ttatgccatg cgaaaaattt tggctgggat aaggcacagc gcgatgtaat
     3061 gagcaaaata tattttacca ctatgttact ttcctttttg gtactccttg catatggttt
     3121 atggtctaaa tcctcgttgg aagatttttt gttttatatt gttttcatgc tcccgttttt
     3181 tagacatgtg ataatgttat acgttgaaaa caacaaatct attaatcgaa ttatgcgtgt
     3241 taaagatttt attgagaaaa aaatacagaa tataaagata agtggcatta taaacgatga
     3301 catcttatct catgatttaa gatcgattca ggatgaggtt tatgcccatc gatcgacctc
     3361 gaatcctgtg cccaattgtc tccacagatt catgcggaag aataatgagg ctatttatga
     3421 tgattatttt gaagagaact tgaaaatttt accccgatag taagaattcc caggcatcaa
     3481 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     3541 aacgctctcc agctggtacc atatggcggg caggacgccc gccataaact gccaggcatc
     3601 aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt
     3661 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3721 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3781 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3841 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3901 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     3961 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     4021 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4081 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4141 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4201 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4261 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4321 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4381 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4441 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4501 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4561 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4621 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4681 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4741 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4801 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4861 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4921 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4981 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5041 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5101 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5161 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5221 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5281 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5341 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5401 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5461 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     5521 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     5581 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgc
//
