LOCUS       sequence-444242-        5636 bp    DNA     circular SYN 15-APR-2025
DEFINITION  For expression of the type I CBASS system encoded by Alteromonas sp.
            ALT199.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5636)
  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 5636)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_Alteromonas_sp._ALT199
FEATURES             Location/Qualifiers
     source          1..5636
                     /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      3475..3546
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3607..3634
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3653..3744
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3745..3813,3814..4605)
                     /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(3963..3982)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4776..5364
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5265..5284
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5518..5535
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(5550..5636,1..54)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5616..5635)
                     /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 tttccatacc cgttttttgg gctaacagga ggaattaacc atggactgct caactttact
     1561 atatagtcaa aacaaagaat taaacaccct tcacaatagg ttaaaaatat ctgaagttgt
     1621 tttaacgaaa ggtgtctata aaaaaaacga gctaattgat tttcttagag aagagcttaa
     1681 agaaacattt gattgcgaag tcaggttttg gcttcaaggc tcctacaaaa gtcatacact
     1741 tataaaacct tctgataaat attcttcata tgatattgat attgggattt atctatcctt
     1801 tgaaatagag gaagctgatg catctgaagt aaaacaggct gtaaaggaag cgctagagtc
     1861 tttctgtaag gttaatgatg aatgtaatct acagcaatca aaaaacgcat gtgaaggact
     1921 taagtataaa cagttgctta ccattgatac tcctgtatat gcattctcta gtacagctat
     1981 taagttagca actgatgatg gttgggtgga tagtgacccg aagttaattc aagactattt
     2041 gactaatgcg ttcgagaatc ccgttgatcg tgcaattatg aaaagggttg ttagatacct
     2101 gaaggcttgg atcaatgtaa agtggaatgg aactgattat aagaaaatac cgtcattggc
     2161 agtaaatatc ctcgttgctg accatttaat aattgaagag aacgaggacg acacttttat
     2221 taacactgtt ttatccattt gcgagtctct agataaagta tttcgagtta atagccctat
     2281 tgatagcagt aaccttcttt caatgccaga tgaagcgata tcttttgctt ataaaaaact
     2341 tgatgaacta aaaagaacct gtaataaatg cgttaagtct cctgactaca gacctttatt
     2401 gctagcaaat ttatttgagc attattttcc accgctaact gaagagagtg gttcaggaag
     2461 catcaattta cctgcattgt caaaagttcc gaacatatcg atcagtcgct attcaaaagg
     2521 cgataaacat attgagacag ttgatgagaa tgaaatcaca gtatttaaag gagaatcttt
     2581 aaccttcagt atctgtaata aaaacgattt taactctgaa tcaacagttc attggacggt
     2641 taggaatatc ggtaagcaag cagatgacgc caacgatatt gggcataaaa atatatcaaa
     2701 tttagaaagc aatgagcgcc gagatgcaac atatacaggc tcgcacataa tggagtgtat
     2761 gatcgtggct aacggcagaa ttcaaggaat tagttctgtg ttagtagagg ttaaaccctc
     2821 aaaagttgta tctagaagga ggaagaaaat atttaaagga tttaggaggt agattatgtg
     2881 ggaagtaata cctagcaagg ttaaatacgc aattgtagtg gtagtttcag ctttatgttt
     2941 ttttctattc ttttcagtat gcgaattcag tttttttagg tctatacttt cggctatttc
     3001 tttagtaaca gtgttagcgc tagtaatcgg aaagtacata tggaaatact tgtattttga
     3061 cgcgctaaat aaatatcttt gccctgacct gaatgggaat tggagcgcca agattaaatc
     3121 taattttggc aaagctactg acgtagttat tcctattaag atagaggctg attttttttc
     3181 cgtgagaatg ttcgcagata cttcttttgg acaatccgaa gccaattact gtcgaatttt
     3241 caaaagtgaa agcgatagat ttcaattgga gtatatgttt caagtcagaa acgattccgc
     3301 tagagaaggg gatgctcaat tttatgaggg tgctgctaga ttgaccctca atgacgcaga
     3361 gaatcagata tggaaaggtg tttactggac aaacagatgt tggaataagg ggaaaaatac
     3421 tgctggagaa atagagcttt caaggccaga aaagtaataa gaattcccag gcatcaaata
     3481 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     3541 gctctccagc tggtaccata tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa
     3601 ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact cttttgttta
     3661 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3721 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     3781 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     3841 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     3901 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     3961 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     4021 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     4081 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     4141 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     4201 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     4261 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     4321 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     4381 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     4441 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     4501 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     4561 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4621 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4681 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4741 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4801 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4861 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4921 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4981 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5041 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5101 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5161 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5221 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5281 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5341 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5401 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5461 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     5521 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     5581 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgc
//
