LOCUS       sequence-444234-        5940 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of the type I CBASS system encoded by Rhodanobacter 
            denitrificans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5940)
  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 5940)
  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_Rhodanobacter_denitrificans
FEATURES             Location/Qualifiers
     source          1..5940
                     /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      3779..3850
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3911..3938
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3957..4048
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4049..4117,4118..4909)
                     /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(4267..4286)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5080..5668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5569..5588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5822..5839
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(5854..5940,1..54)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5920..5939)
                     /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 atgagcacgt ccgcagtggc
     1561 tatcgggcag ggttcggaac gcccacctaa ccagcgattc cagattcttg ccctctccgg
     1621 cgggggttac cgcgggttat ataccgcgaa gattttggcc gacctcgaac aacatatcgg
     1681 ggcgcccatt ggccgccatt tcgatctcat cgccggcaca tccatcgggg gcatcctggc
     1741 cttggcggtg gcgctggaaa ttcccgcgga gcgtatggtg acgcttttcg agcggcacgg
     1801 ggaagcgatc tttcggcgtc ggtggtcgtt gcgcggcatc gtccgcgccc cttactcgca
     1861 ggcaccgctg gccgcgttgc tggcgcagga tgacctgttc ggtgatcgcc gtttggaggc
     1921 gtgccttcac ccggtgctgg tacccaccat caactacagc actggcctcc cggtgctctt
     1981 caagacaccg catcacccga atttctcccg cgatttccgc taccagctga ttgatgtggc
     2041 cttggccacc agtgcagccc ccgcatactt tccgcgccac gtcttcgatc atcgtcagta
     2101 catcgatggc ggactctttg ccaacgcccc gggtttgctc gccctgcatg aggcccaaca
     2161 tttcctcggc cgcccccggg aagacatctg cctagtcgcc atcggcacca tgtccgcgcg
     2221 attcaccgtt gacccacggc gcaatcggag tggtggcacc tacgactggg gcggctggca
     2281 cccggcggaa acgcccaagc ggttgtttgg cgtcgccatc tccgcgcagg aatcgctggt
     2341 ccatcacctg ctgggccatc agctggctcc tggccaatac ttccacgtgg atgacgacct
     2401 gaccgatgca cgtgctcgcg cggtggcact cgacaaggcc aacgcggccg cacgcgaagt
     2461 gctgcttggt gtgggcgccg aacgcgcgaa atggtgtctt ggtgacccag cttttcggac
     2521 cgtcctccaa cactccccca tcgccccccg cttctaccac ggtatccatg cccaccacga
     2581 agaaggagtc gctcgcgatg ctgtcactgc ataagttgtt tttgaacgcg ggcgcgcccg
     2641 acagcttcga tgaggtgatt gcgccgacgg atagtcagcg tcggacgctg atggccgcca
     2701 aaaatgccat tcgcgagcat cttcgcgcgg acattcgggc agctacaact accgtgcttg
     2761 gcatggatcg catggtgacg ccgcgctttc gcactcaggg ttcgtgggcc tacaagacct
     2821 gcgtgcaagc cgcccaccgc ccaccgcagg agatggactg ggatttcggc gtctatctgc
     2881 cggtgaccgt gtgggacgag catgccccgc cggcccgcat ggccaagttg tattttgagc
     2941 tggtggaacg ttcgctgggc caattgtgcg aacgcgaggg ctggcgactg gagcgtggca
     3001 acacacgctg cgttcggatc cgaattgccg attgggctca catcgatgtg cccctgtatg
     3061 ccgcccccga ggccaagttt caccaggtaa tggaaaaagc cgtggcgttg aatagtgccc
     3121 accatgccta ccttcaccta cgcgaaagcg cggcgctgga cgaagcgccg gagcccttct
     3181 gggagctgct ggacgagatc catgtagcaa cgcgtgacgg ccgctggctg ccgtccgatc
     3241 cggaggccgt ggcgaagtgg ttcgatgatc aagtgctggt gcatggcgag cagctgcggc
     3301 gtgtgtgctg ctacctcaag gcatggcgcg actatcactg gcgcgatggc ggcggtccga
     3361 gttcggtatt gatcatgatc gtggttactc aagcgtttga cgccaccccg cgacgcgatg
     3421 atctggccgt cgagcgggcc gccatgacac tcgcccgggc gttgggcgac ggggtgtacg
     3481 agcgcggcat tgacggcggc gaggaggatt ttaaccgcat gtcatccggc gagcgcctga
     3541 tcgcggtgca gcgggcggag tcgctggccc gtcagatgtc ccttagccgc cacttcgggc
     3601 cgggcctcag gcagacggct gtcgataacg tgcgcgcgca gtttggcccg cgcatcgccg
     3661 ataactgcga actggtggca gccgacgatg gcaccgacgt gcgccgcgtg ccggccgccc
     3721 aagtggtgcc tccggtggtg ggtgccacga aggccggcta ataagaattc ccaggcatca
     3781 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     3841 gaacgctctc cagctggtac catatggcgg gcaggacgcc cgccataaac tgccaggcat
     3901 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg
     3961 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4021 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4081 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4141 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4201 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4261 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     4321 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4381 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4441 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4501 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4561 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4621 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4681 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4741 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4801 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4861 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4921 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4981 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5041 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5101 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5161 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5221 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5281 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5341 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5401 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5461 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5521 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5581 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5641 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5701 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5761 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     5821 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     5881 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
//
