LOCUS       sequence-488906-        4795 bp    DNA     circular SYN 09-OCT-2025
DEFINITION  Bacterial expression of N4gp48 under control of the pJex promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4795)
  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 4795)
  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: pBA1866
FEATURES             Location/Qualifiers
     source          1..4795
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      502..573
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      589..616
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1207..2157)
                     /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      2205..2427
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(2970..3785)
                     /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(2970..3785)
                     /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     3693..3712
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3777..3796
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     RBS             4554..4562
                     /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 actgatgaac
      121 cgtattactg aagaacatat tgatagcatc attgaaggtg tttactactt caatatggct
      181 aaagctgtag agaatgaagc agttctgatt cctgaggacc atttaaaagc attaagtaca
      241 actactgttt gtgcactggt tctttctaat ggattcacta ttactggtaa atctgcctgt
      301 gttgacccta acaccttcaa tgaggagtta ggtaagaaat atgcttacga acaggccgtt
      361 gataaagtat gggaattgga aggctatctg cttaagcaaa acctgcatga gaaaactcag
      421 agccaggaaa tgctaaaagg gttcctgcaa gatgacaact gtgaaggtgg gggctgtaaa
      481 atctaaagga tctccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
      541 cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg
      601 ggtgggcctt tctgcgttta tagaagacca aggaggactt tggcaggtgc tctcctgagt
      661 aggacaaatc cgccgcccta gacctagggt acgggttttg ctgcccgcaa acgggctgtt
      721 ctggtgttgc tagtttgtta tcagaatcgc agatccggct tcagccggtt tgccggctga
      781 aagcactatt tcttccagaa ttgccatgat tttttcccca cgggaggcgt cactggctcc
      841 cgtgttgtcg gcagctttga ttcgataagc agcatcgcct gtttcaggct gtctatgtgt
      901 gactgttgag ctgtaacaag ttgtctcagg tgttcaattt catgttctag ttgctttgtt
      961 ttactggttt cacctgttct attaggtgtt acatgctgtt catctgttac attgtcgatc
     1021 tgttcatggt gaacagcttt gaatgcacca aaaactcgta aaagctctga tgtatctatc
     1081 ttttttacac cgttttcatc tgtgcatatg gacagttttc cctttgatat gtaacggtga
     1141 acagttgttc tacttttgtt tgttagtctt gatgcttcac tgatagatac aagagccata
     1201 agaacctcag atccttccgt atttagccag tatgttctct agtgtggttc gttgtttttg
     1261 cgtgagccat gagaacgaac cattgagatc atacttactt tgcatgtcac tcaaaaattt
     1321 tgcctcaaaa ctggtgagct gaatttttgc agttaaagca tcgtgtagtg tttttcttag
     1381 tccgttatgt aggtaggaat ctgatgtaat ggttgttggt attttgtcac cattcatttt
     1441 tatctggttg ttctcaagtt cggttacgag atccatttgt ctatctagtt caacttggaa
     1501 aatcaacgta tcagtcgggc ggcctcgctt atcaaccacc aatttcatat tgctgtaagt
     1561 gtttaaatct ttacttattg gtttcaaaac ccattggtta agccttttaa actcatggta
     1621 gttattttca agcattaaca tgaacttaaa ttcatcaagg ctaatctcta tatttgcctt
     1681 gtgagttttc ttttgtgtta gttcttttaa taaccactca taaatcctca tagagtattt
     1741 gttttcaaaa gacttaacat gttccagatt atattttatg aattttttta actggaaaag
     1801 ataaggcaat atctcttcac taaaaactaa ttctaatttt tcgcttgaga acttggcata
     1861 gtttgtccac tggaaaatct caaagccttt aaccaaagga ttcctgattt ccacagttct
     1921 cgtcatcagc tctctggttg ctttagctaa tacaccataa gcattttccc tactgatgtt
     1981 catcatctga gcgtattggt tataagtgaa cgataccgtc cgttctttcc ttgtagggtt
     2041 ttcaatcgtg gggttgagta gtgccacaca gcataaaatt agcttggttt catgctccgt
     2101 taagtcatag cgactaatcg ctagttcatt tgctttgaaa acaactaatt cagacataca
     2161 tctcaattgg tctaggtgat tttaatcact ataccaattg agatgggcta gtcaatgata
     2221 attactagtc cttttccttt gagttgtggg tatctgtaaa ttctgctaga cctttgctgg
     2281 aaaacttgta aattctgcta taccctctgt aaattccgct agacctttgt gtgttttttt
     2341 tgtttatatt caagtggtta taatttatag aataaagaaa gaataaaaaa agataaaaag
     2401 aatagatccc agccctgtgt ataactcact actttagtca gttccgcagt attacaaaag
     2461 gatgtcgcaa acgctgtttg ctcctctaca aaacagacct taaaacccta aaggcttaag
     2521 tagcaccctc gcaagctcgg gcaaatcgct gaatattcct tttgtctccg accatcaggc
     2581 acctgagtcg ctgtcttttt cgtgacattc agttcgctgc gctcacggct ctggcagtga
     2641 atgggggtaa atggcactac aggcgccttt tatggattca tgcaaggaaa ctacccataa
     2701 tacaagaaaa gcccgtcacg ggcttctcag ggcgttttat ggcgggtctg ctatgtggtg
     2761 ctatctgact ttttgctgtt cagcagttcc tgccctctga ttttccagtc tgaccacttc
     2821 ggattatccc gtgacaggtc attcagactg gctaatgcac ccagtaaggc agcggtatca
     2881 tcaacaggct tacccgtctt actgtccctg tgtcagattt cttactttat tcaggcgatt
     2941 acgtgttaca accaattaac caattctgat tagaaaaact catcgagcat caaatgaaac
     3001 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     3061 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     3121 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     3181 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc
     3241 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     3301 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga ggcgaaatac gcgatcgctg
     3361 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     3421 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     3481 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     3541 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     3601 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     3661 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     3721 tcagcatcca tgttggaatt taatcgcggc ctggagcaag acgtttcccg ttgaatatgg
     3781 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat
     3841 atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttgttga
     3901 ataaatcgaa cttttgctga gttgaaggat cagcacctgc ccgaaccaga gacttatctc
     3961 aggacgtctt acgaaaataa ctcaagctga ataacgtgct gcagatggcg ggtgaatgcg
     4021 gcatcgtcta cctccggcat tcccagcacg taaatcccgt ccagtccaca gaccagcgaa
     4081 atcagccgcc aggcgatatt ttcggcgcta tcgcgcaggg taaattcgcc agcggcatgg
     4141 cccgcgcgaa tgatcctgac cgcttcgtca tgccacaggt tcatggtcag caggtacgcg
     4201 cttttgattt ccggatcgct gtcggccagc agctgcgcct ggcgccacag gcggatatag
     4261 ggctccagcc gtccgtcctc gctgccgagg gcggaaaaca gctgttcacg ccagccggcg
     4321 gtgcgcgaaa ggcgctgcag atccatcatc tcgcggatca cgcggataaa cgcctgggat
     4381 tttagctcgc cggaggaggt gaaatggtgg tggacctggc ctgcggcgac gcccgcggcg
     4441 gtggcgatgt tgcggaccgt cataccggta aacccttgat ccagcgcgac gcgcatggcg
     4501 gcctgcatga tggtttcccg acgttcttcg cgattcagat agcccatatt gtcacctcct
     4561 tatccacaca ttatacgagc cggatgatta atggggaacc ttaccagagg gcgccccagc
     4621 tggcaattcc tcttaagtaa gtaagagtat acgtatatcg gctaataacg tattaaggcg
     4681 cttcggcgcc tttttttatg ggggtatttt catcccaatc cacacgtcca acgcacagca
     4741 aacaccacgt cgaccctatc agctgcgtgc tttctatgag tcgttgctgc ataac
//
