LOCUS       sequence-444244-        5635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of the type I CBASS system encoded by 
            Pseudoalteromonas sp. A2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5635)
  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 5635)
  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_Pseudoalteromonas_sp._A2
FEATURES             Location/Qualifiers
     source          1..5635
                     /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      3474..3545
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3606..3633
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3652..3743
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3744..3812,3813..4604)
                     /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(3962..3981)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4775..5363
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5264..5283
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5517..5534
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(5549..5635,1..54)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5615..5634)
                     /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 atgagttgta atgaactatt
     1561 tttttctagt aatgataaag atgaaactct tcaccaaaga acacaattat cagatgttgt
     1621 tttgtcaaaa ggcattacta aaaaggatga gttaattgaa tttcttcgat ctgaattaag
     1681 tgaagccttt gaatgtaaag ttgggttttg gttacaaggt tcttataaaa gtcgtacatt
     1741 aattaaacca gtagataaat tttcttccta tgatatcgat atcggtgttt atttgttttt
     1801 tgacgcagaa tctgaagaga ttgattctaa ggatgtgaaa tcaacactaa aagatgcctt
     1861 gtcttcatat tttgaaataa acgatgaagc agagcctcaa aaatcaaaaa acgcttgcga
     1921 aggactaaaa tattcgacat tccttactat tgatacacct atttatttta ttgatggctc
     1981 taatattaaa ttagctacgg atgacgggtg ggttgaaagt gatccaaaag ctattcagga
     2041 ttggcttacg aactactata gcgaaaaagc ggatcgagca ttgatgaagc gtttggttcg
     2101 ctacttcaaa gcttggatta atgttaaatg gcaaggtaca gggtttaaaa aaataccctc
     2161 gatagcagtt aacgttttag tcgcacagca catgcaaaag catgatagag aagatgactc
     2221 ttttcttcat acagctttaa gtatatgtga agaactagag tccacattta tagtaaataa
     2281 cccattgggt ggaagtaacc tcctgacaat gcctgaagac tcagaaagtt ttgctcatca
     2341 gaaacttgat gaattaaaag ctatatgcct taactgtatt gattcaagtg aatttgatag
     2401 aggaattaat ttttcaaacc tttttcaaca ttacttccct caagtaaata taagttcttc
     2461 tgagggaaca acaggtttac ctgccgtagt tactgtacct gaaatatcaa tttgcagata
     2521 tgacaaagat ggaaatcatg ttgaaacaac taccactgat aatttaacag ttaaaaaagg
     2581 tgactctctt acctttacta tatgtaatca agattcattt aatacttatt cagatgcaca
     2641 atggacagtt aggaatgtag gcggtcaagc aacagatgct aatgatattg ggcataaggt
     2701 tattgggagg ctaaatgaaa gccataaaag ggatacatca tatacaggct cacacactat
     2761 ggagtgtatg attactcata acggagtggt aaaaggtttt caggctattc atgtccaagt
     2821 gaagccagcc agtaccctaa gccgaaaaaa aacatttaaa ggttggaggc gctaattatg
     2881 tgggatcttt ttccatcaaa ggttaagtat ggtctcccag ctgtaattgc aattctaatt
     2941 tatttatttt gtaatcaata tctcgatcaa ccattaatta gatctatttc ttatacgatt
     3001 acagctataa caattttagc ttggcttttt ggtaaatatc tttggaagtt tgtatacata
     3061 gactttttca aaaacaaatt ttgtcctgat tttaatggca aatggatagg tactattgag
     3121 tcaaatttta atggtggcac caccgtccag ttcccaatta caattgaagc tgatttcttt
     3181 tcaattagaa tgaaaggtaa caccacagta ggtagaactt acgcaaatta ctgtaaggta
     3241 atcagaactg atgatgattg ttttgaactt gaatatatgt ttaaagggat aaatgatacg
     3301 ccgtccgaaa ctgatactag cttctatgag ggggcagcta ggcttagagt tctagaaaca
     3361 tcgactatgg aaatgaaagg ggtattctgg actaatagat gttggcaaaa cggaaaaaat
     3421 acagcaggtg tattagagct taagaaagac tcataataag aattcccagg catcaaataa
     3481 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     3541 ctctccagct ggtaccatat ggcgggcagg acgcccgcca taaactgcca ggcatcaaat
     3601 taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat
     3661 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3721 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3781 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3841 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3901 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3961 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     4021 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4081 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4141 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4201 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4261 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4321 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4381 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4441 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4501 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4561 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4621 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4681 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4741 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4801 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4861 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4921 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4981 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5041 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5101 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5161 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5221 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5281 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5341 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5401 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5461 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     5521 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     5581 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgc
//
