LOCUS       sequence-444233-        5632 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of the type I CBASS system encoded by Escherichia 
            albertii.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5632)
  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 5632)
  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_albertii
FEATURES             Location/Qualifiers
     source          1..5632
                     /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      3471..3542
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3603..3630
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3649..3740
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3741..3809,3810..4601)
                     /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(3959..3978)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4772..5360
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5261..5280
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5514..5531
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(5546..5632,1..54)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5612..5631)
                     /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 atgaattgca gtgacctttt
     1561 ttatgctgat acaaatacag aaaacacact tcatcagaga actcaattat ctgaagttat
     1621 attatctaaa ggtattgcaa agaaaaatga gttaatagaa tttctaagac aagagttaaa
     1681 ggaagcgttt gactgtgatg tgcgattttg gttacaaggt tcatataaaa gccatacgct
     1741 gataaaaccg gtagataagt tttcatcata tgatatcgat atcggcgtat atttattttt
     1801 cgatgctgaa aatgaggggg ttgattctaa agatgttaag gaaacgctga gggatgcact
     1861 attgtcttat tgttccatta ataatgaagc aaaactgcaa gagtcaaaaa atgcttgtga
     1921 aggactaaag ttctctactt ttcttactgt tgatactcct atttattata aaacagatac
     1981 taagataaaa ttagcgacag acaaaggctg gagtgacagt gatccaaaag ctattcagga
     2041 ttggattaca aattattata aagacaaatc tgacagagct ttaatgaagc gactcgttcg
     2101 atacttcaaa gcttgggtaa atgtaaagtg gcaaaacacc gggtttaaaa aaataccctc
     2161 attagctatt aatgttttgg tagcccagca tatgaaacag catgtgcgag aagatgattg
     2221 ttttatatat acggcgttaa gtatttgcga ggaactagaa tctacattaa tagttagaaa
     2281 ccctttaaat aatagcaact taatttccat gcctcaagat gctgagtgct ttgcacatca
     2341 aaaacttgat gaattaaagc aagtatgcct tagctgcatt aagtccgatg atattaaaag
     2401 gggagcccat ttttcaaatc ttttccaaca ttacttccca caaatatcat tagattctgc
     2461 tacaggtagc actggtctgc ctacggtagt taatgttcct gaaatctcag tttgtagata
     2521 tgataaaaat ggtaatcatg ttgaaacaat aattactgat agattgactg ttaataaagg
     2581 ggattcacta acttttacaa tccgtaatca ttatgatttt aatatctatt ctagtgcgca
     2641 atggacagtc agaaatattg gctcacaagc aaatgatgct aatgatattg gacactcagt
     2701 cacaggtaaa cctagtgaaa gtcataaacg aggcacttcg tatacggggt ctcatactat
     2761 ggaatgtatg attttacata atggggcaat aataggattt aaaactatac atgtaatagt
     2821 aaaacctgcg cgtacagtaa gaagaaaaac acttaaattc tggagggcat gagaatgtgg
     2881 gagcttttac catcgaagat aaaatatggg atatctacta taatatctat tattgttttt
     2941 ttgttttttt tggagtttct cgggcaacca gttttgaagt cggtttctta tactattact
     3001 actattacga ttttggcttt tatatttggt aagtatttat ggaagtattt ttatatagac
     3061 tatttaaaac ataaattctg ccctgatttt aatgggcgat ggataggtaa aatagagtca
     3121 aattacagtg gtggaactaa ggttgagttt ccactggaga ttaaagctga ttttttttca
     3181 attaaaatga aaggaaatac cacaataggg agaacttact caaattactg taaggttgta
     3241 agggctgaag atgatagttt tgaacttgta tacatgttta aagttttcaa tgatacacct
     3301 tcaataactg atactagctt ttacgaaggt gcagccagat tacgagtgat tgatattaag
     3361 acaatgaaca tgaaaggggt gttttggact aatcgatgtt gggaaaatgg caagaataca
     3421 gcaggtataa ttgaactttc aaaagaagtt taataagaat tcccaggcat caaataaaac
     3481 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     3541 tccagctggt accatatggc gggcaggacg cccgccataa actgccaggc atcaaattaa
     3601 gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt
     3661 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3721 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3781 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3841 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3901 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3961 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     4021 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4081 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4141 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4201 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4261 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     4321 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4381 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4441 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4501 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4561 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     4621 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4681 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4741 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4801 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4861 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     4921 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4981 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5041 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5101 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5161 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5221 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5281 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5341 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5401 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5461 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     5521 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     5581 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gc
//
