LOCUS       sequence-488905-        4882 bp    DNA     circular SYN 09-OCT-2025
DEFINITION  Bacterial expression of T4 Acb1 under control of the pJex promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4882)
  AUTHORS   Doherty EE, Adler BA, Yoon PH, Hsieh K, Loi K, Armbuster EG, Lahiri 
            A, Bolling CS, Wilcox XE, Akkati A, Iavarone AT, Pogliano J, Doudna 
            JA
  TITLE     A miniature CRISPR-Cas10 enzyme confers immunity by an inverse 
            signaling pathway.
  JOURNAL   bioRxiv [Preprint]. 2025 Apr 1:2025.03.28.646030. doi: 
            10.1101/2025.03.28.646030.
  PUBMED    40196485
REFERENCE   2  (bases 1 to 4882)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 9, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2025 Apr 1:2025.03.28.646030. doi: 
            10.1101/2025.03.28.646030."
COMMENT     Sequence Label: pBA1398
FEATURES             Location/Qualifiers
     source          1..4882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      589..660
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      676..703
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1294..2244)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2292..2514
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(3057..3872)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3057..3872)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3780..3799
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3864..3883
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     RBS             4641..4649
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
ORIGIN
        1 aaaaaactgc aaaaaatagt ttgacaggac acgtgtccaa ctttaagatg tacccaaagt
       61 tggacacgtg tccaactttg aattcaaaag atcttttaag aaggagatat actgatgatg
      121 gaatttaaag acttttcaac gggtctttat gtagcagcta agttttcaga attaacactt
      181 gatgcgctgg aagaacttca gcgctcttta cgtgttccta atccagttcc tagagaaaaa
      241 attcattcga ctatatgtta ttcaagagta aatgttccat atgttccatc gagtggaagt
      301 tttgaagtag cttcttctgg acatttagaa gtatggaaaa cacaagatgg atcgactctt
      361 gtacttgtgc tagattctga atatctgcgc tgtcgacaca tgtatgcgcg ggcactaggt
      421 gctacacatg attttgatga ttacacaccg catataacat tgtcttataa tgttgggccc
      481 ttatcattta gcggtgatgt acaaattccg gtcgtattag atcgtgaata caaagagcct
      541 cttaaactcg attgggcaga tgatttaaaa taaaggatct ccaggcatca aataaaacga
      601 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
      661 tactagagtc acactggctc accttcgggt gggcctttct gcgtttatag aagaccaagg
      721 aggactttgg caggtgctct cctgagtagg acaaatccgc cgccctagac ctagggtacg
      781 ggttttgctg cccgcaaacg ggctgttctg gtgttgctag tttgttatca gaatcgcaga
      841 tccggcttca gccggtttgc cggctgaaag cactatttct tccagaattg ccatgatttt
      901 ttccccacgg gaggcgtcac tggctcccgt gttgtcggca gctttgattc gataagcagc
      961 atcgcctgtt tcaggctgtc tatgtgtgac tgttgagctg taacaagttg tctcaggtgt
     1021 tcaatttcat gttctagttg ctttgtttta ctggtttcac ctgttctatt aggtgttaca
     1081 tgctgttcat ctgttacatt gtcgatctgt tcatggtgaa cagctttgaa tgcaccaaaa
     1141 actcgtaaaa gctctgatgt atctatcttt tttacaccgt tttcatctgt gcatatggac
     1201 agttttccct ttgatatgta acggtgaaca gttgttctac ttttgtttgt tagtcttgat
     1261 gcttcactga tagatacaag agccataaga acctcagatc cttccgtatt tagccagtat
     1321 gttctctagt gtggttcgtt gtttttgcgt gagccatgag aacgaaccat tgagatcata
     1381 cttactttgc atgtcactca aaaattttgc ctcaaaactg gtgagctgaa tttttgcagt
     1441 taaagcatcg tgtagtgttt ttcttagtcc gttatgtagg taggaatctg atgtaatggt
     1501 tgttggtatt ttgtcaccat tcatttttat ctggttgttc tcaagttcgg ttacgagatc
     1561 catttgtcta tctagttcaa cttggaaaat caacgtatca gtcgggcggc ctcgcttatc
     1621 aaccaccaat ttcatattgc tgtaagtgtt taaatcttta cttattggtt tcaaaaccca
     1681 ttggttaagc cttttaaact catggtagtt attttcaagc attaacatga acttaaattc
     1741 atcaaggcta atctctatat ttgccttgtg agttttcttt tgtgttagtt cttttaataa
     1801 ccactcataa atcctcatag agtatttgtt ttcaaaagac ttaacatgtt ccagattata
     1861 ttttatgaat ttttttaact ggaaaagata aggcaatatc tcttcactaa aaactaattc
     1921 taatttttcg cttgagaact tggcatagtt tgtccactgg aaaatctcaa agcctttaac
     1981 caaaggattc ctgatttcca cagttctcgt catcagctct ctggttgctt tagctaatac
     2041 accataagca ttttccctac tgatgttcat catctgagcg tattggttat aagtgaacga
     2101 taccgtccgt tctttccttg tagggttttc aatcgtgggg ttgagtagtg ccacacagca
     2161 taaaattagc ttggtttcat gctccgttaa gtcatagcga ctaatcgcta gttcatttgc
     2221 tttgaaaaca actaattcag acatacatct caattggtct aggtgatttt aatcactata
     2281 ccaattgaga tgggctagtc aatgataatt actagtcctt ttcctttgag ttgtgggtat
     2341 ctgtaaattc tgctagacct ttgctggaaa acttgtaaat tctgctatac cctctgtaaa
     2401 ttccgctaga cctttgtgtg ttttttttgt ttatattcaa gtggttataa tttatagaat
     2461 aaagaaagaa taaaaaaaga taaaaagaat agatcccagc cctgtgtata actcactact
     2521 ttagtcagtt ccgcagtatt acaaaaggat gtcgcaaacg ctgtttgctc ctctacaaaa
     2581 cagaccttaa aaccctaaag gcttaagtag caccctcgca agctcgggca aatcgctgaa
     2641 tattcctttt gtctccgacc atcaggcacc tgagtcgctg tctttttcgt gacattcagt
     2701 tcgctgcgct cacggctctg gcagtgaatg ggggtaaatg gcactacagg cgccttttat
     2761 ggattcatgc aaggaaacta cccataatac aagaaaagcc cgtcacgggc ttctcagggc
     2821 gttttatggc gggtctgcta tgtggtgcta tctgactttt tgctgttcag cagttcctgc
     2881 cctctgattt tccagtctga ccacttcgga ttatcccgtg acaggtcatt cagactggct
     2941 aatgcaccca gtaaggcagc ggtatcatca acaggcttac ccgtcttact gtccctgtgt
     3001 cagatttctt actttattca ggcgattacg tgttacaacc aattaaccaa ttctgattag
     3061 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     3121 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     3181 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     3241 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     3301 tccggtgaga atggcaaaag cttatgcatt tctttccaga cttgttcaac aggccagcca
     3361 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     3421 tgagcgaggc gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc
     3481 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     3541 tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca
     3601 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
     3661 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     3721 tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta
     3781 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctg
     3841 gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa
     3901 gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga
     3961 ttttgagaca caacgtggct ttgttgaata aatcgaactt ttgctgagtt gaaggatcag
     4021 cacctgcccg aaccagagac ttatctcagg acgtcttacg aaaataactc aagctgaata
     4081 acgtgctgca gatggcgggt gaatgcggca tcgtctacct ccggcattcc cagcacgtaa
     4141 atcccgtcca gtccacagac cagcgaaatc agccgccagg cgatattttc ggcgctatcg
     4201 cgcagggtaa attcgccagc ggcatggccc gcgcgaatga tcctgaccgc ttcgtcatgc
     4261 cacaggttca tggtcagcag gtacgcgctt ttgatttccg gatcgctgtc ggccagcagc
     4321 tgcgcctggc gccacaggcg gatatagggc tccagccgtc cgtcctcgct gccgagggcg
     4381 gaaaacagct gttcacgcca gccggcggtg cgcgaaaggc gctgcagatc catcatctcg
     4441 cggatcacgc ggataaacgc ctgggatttt agctcgccgg aggaggtgaa atggtggtgg
     4501 acctggcctg cggcgacgcc cgcggcggtg gcgatgttgc ggaccgtcat accggtaaac
     4561 ccttgatcca gcgcgacgcg catggcggcc tgcatgatgg tttcccgacg ttcttcgcga
     4621 ttcagatagc ccatattgtc acctccttat ccacacatta tacgagccgg atgattaatg
     4681 gggaacctta ccagagggcg ccccagctgg caattcctct taagtaagta agagtatacg
     4741 tatatcggct aataacgtat taaggcgctt cggcgccttt ttttatgggg gtattttcat
     4801 cccaatccac acgtccaacg cacagcaaac accacgtcga ccctatcagc tgcgtgcttt
     4861 ctatgagtcg ttgctgcata ac
//
