LOCUS       sequence-491110-        5847 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of Panoptes locus with 300bp of context 
            upstream and 150bp downstream of the ECOR31 operon cloned under the 
            pJex promoter; SC101 ori.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5847)
  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 5847)
  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"; journalName: "bioRxiv 
            [Preprint]. 2025 Apr 1:2025.03.28.646030. doi: 
            10.1101/2025.03.28.646030."
COMMENT     Sequence Label: pBA1751
FEATURES             Location/Qualifiers
     source          1..5847
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      253..475
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(1018..1833)
                     /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(1018..1833)
                     /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     1741..1760
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1825..1844
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     RBS             2602..2610
                     /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)"
     terminator      4397..4468
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4484..4511
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(5102..5847,1..205))
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 caccataagc attttcccta ctgatgttca tcatctgagc gtattggtta taagtgaacg
       61 ataccgtccg ttctttcctt gtagggtttt caatcgtggg gttgagtagt gccacacagc
      121 ataaaattag cttggtttca tgctccgtta agtcatagcg actaatcgct agttcatttg
      181 ctttgaaaac aactaattca gacatacatc tcaattggtc taggtgattt taatcactat
      241 accaattgag atgggctagt caatgataat tactagtcct tttcctttga gttgtgggta
      301 tctgtaaatt ctgctagacc tttgctggaa aacttgtaaa ttctgctata ccctctgtaa
      361 attccgctag acctttgtgt gttttttttg tttatattca agtggttata atttatagaa
      421 taaagaaaga ataaaaaaag ataaaaagaa tagatcccag ccctgtgtat aactcactac
      481 tttagtcagt tccgcagtat tacaaaagga tgtcgcaaac gctgtttgct cctctacaaa
      541 acagacctta aaaccctaaa ggcttaagta gcaccctcgc aagctcgggc aaatcgctga
      601 atattccttt tgtctccgac catcaggcac ctgagtcgct gtctttttcg tgacattcag
      661 ttcgctgcgc tcacggctct ggcagtgaat gggggtaaat ggcactacag gcgcctttta
      721 tggattcatg caaggaaact acccataata caagaaaagc ccgtcacggg cttctcaggg
      781 cgttttatgg cgggtctgct atgtggtgct atctgacttt ttgctgttca gcagttcctg
      841 ccctctgatt ttccagtctg accacttcgg attatcccgt gacaggtcat tcagactggc
      901 taatgcaccc agtaaggcag cggtatcatc aacaggctta cccgtcttac tgtccctgtg
      961 tcagatttct tactttattc aggcgattac gtgttacaac caattaacca attctgatta
     1021 gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc
     1081 atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag
     1141 gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat
     1201 taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga
     1261 atccggtgag aatggcaaaa gcttatgcat ttctttccag acttgttcaa caggccagcc
     1321 attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc
     1381 ctgagcgagg cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg
     1441 caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc
     1501 ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc
     1561 aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag
     1621 tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa
     1681 ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt
     1741 atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta atcgcggcct
     1801 ggagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
     1861 agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag
     1921 attttgagac acaacgtggc tttgttgaat aaatcgaact tttgctgagt tgaaggatca
     1981 gcacctgccc gaaccagaga cttatctcag gacgtcttac gaaaataact caagctgaat
     2041 aacgtgctgc agatggcggg tgaatgcggc atcgtctacc tccggcattc ccagcacgta
     2101 aatcccgtcc agtccacaga ccagcgaaat cagccgccag gcgatatttt cggcgctatc
     2161 gcgcagggta aattcgccag cggcatggcc cgcgcgaatg atcctgaccg cttcgtcatg
     2221 ccacaggttc atggtcagca ggtacgcgct tttgatttcc ggatcgctgt cggccagcag
     2281 ctgcgcctgg cgccacaggc ggatataggg ctccagccgt ccgtcctcgc tgccgagggc
     2341 ggaaaacagc tgttcacgcc agccggcggt gcgcgaaagg cgctgcagat ccatcatctc
     2401 gcggatcacg cggataaacg cctgggattt tagctcgccg gaggaggtga aatggtggtg
     2461 gacctggcct gcggcgacgc ccgcggcggt ggcgatgttg cggaccgtca taccggtaaa
     2521 cccttgatcc agcgcgacgc gcatggcggc ctgcatgatg gtttcccgac gttcttcgcg
     2581 attcagatag cccatattgt cacctcctta tccacacatt atacgagccg gatgattaat
     2641 ggggaacctt accagagggc gccccagctg gcaattcctc ttaagtaagt aagagtatac
     2701 gtatatcggc taataacgta ttaaggcgct tcggcgcctt tttttatggg ggtattttca
     2761 tcccaatcca cacgtccaac gcacagcaaa caccacgtcg accctatcag ctgcgtgctt
     2821 tctatgagtc gttgctgcat aacaaaaaac tgcaaaaaat agtttgacag gacacgtgtc
     2881 caactttaag atgtacccaa agttggacac gtgtccaact ttgaattcaa aagatctttt
     2941 aagaaggaga tatactgatt gagaaatctt tagtgtaaag atttaataat tatcctttaa
     3001 gtgtagatta caagcagaac aattgaatta atcgccacat tgaaaatata aaacctcaac
     3061 acacttgatg tatttaacaa gacataggat taaacccatt aaaattaaaa aaatttaaaa
     3121 tacaaacctc gttttggccg ggtataaata caatttaaac cgcaacgttt ttaagtaaat
     3181 aaattcattt gcaactaagc attcagggtt gatataatat aaccacttag ataataatcc
     3241 atgttagttt ggagttaata tgtacatagc gatagatggc gatgatgtgg ggagaaaaat
     3301 aacctcatca tacttatcca actcagaaga gaggttgaca tacataagca acaaattaaa
     3361 tgataccaca aaaaaaatat ctaaaatgct tttatccaat ggatttgaga taattttcca
     3421 agccgcagat ggtgttacag cgaaaacaga caacgaggtc aatcttaact ttgtttttga
     3481 taagataaaa tcctattcct ttgacgaaat tacattttca gctggtgtgg gagcgaatct
     3541 aagagaagcc tatgttgcac tgttaaattc taaaagcaac ggtaagaata tgatttctat
     3601 ttacaaagac atattataat ggtgcgaaat gtttagaata ataagatttg actggttgtt
     3661 aaaaacggta tcactaatct ctttgtttgt cacaatactt tcattttgga tctttaagac
     3721 agttctgtct tctgacttaa gcctgctcaa attgttatca atatcatcat tagcatccat
     3781 tatcataaac ttctttctcc tttcatcaaa gatatcaaga ctaatatggg gaacaataag
     3841 gtttttcaag aaagacatat atcctgactt aaatggaatt tgggttggac atataatcac
     3901 agaaacaaat gatgaatttg aagtccgcgc aaaaataaag caagcactcc ttgtaacaaa
     3961 gatagaaatg cacggcccca cagttaaatc tgttacgcta gaagcaacgc caactaagga
     4021 actggataac aacaaattgt attatgtata taaatcaacc cctaaaaacc ctagttggag
     4081 cgagtatatt ggctcgacta tattcgacgt aattgaaagt aacaatgcgc tccaactttc
     4141 cggtcgatat tatacagaca gaaaatccgt ggggagaata tcaataaaaa gaatatcact
     4201 taacactgac tccgatatat ctttttatta actttcggaa tgtttaatta ccacaatcag
     4261 ccatgttcgt tcttttaaaa tagaaggata attcgcgcta ctgatatatt aatcatggaa
     4321 aaactccttt cagtagccat tttccctaga aaaatacccc tgcatacctc atatactcca
     4381 taggatctcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     4441 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
     4501 gcctttctgc gtttatagaa gaccaaggag gactttggca ggtgctctcc tgagtaggac
     4561 aaatccgccg ccctagacct agggtacggg ttttgctgcc cgcaaacggg ctgttctggt
     4621 gttgctagtt tgttatcaga atcgcagatc cggcttcagc cggtttgccg gctgaaagca
     4681 ctatttcttc cagaattgcc atgatttttt ccccacggga ggcgtcactg gctcccgtgt
     4741 tgtcggcagc tttgattcga taagcagcat cgcctgtttc aggctgtcta tgtgtgactg
     4801 ttgagctgta acaagttgtc tcaggtgttc aatttcatgt tctagttgct ttgttttact
     4861 ggtttcacct gttctattag gtgttacatg ctgttcatct gttacattgt cgatctgttc
     4921 atggtgaaca gctttgaatg caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt
     4981 tacaccgttt tcatctgtgc atatggacag ttttcccttt gatatgtaac ggtgaacagt
     5041 tgttctactt ttgtttgtta gtcttgatgc ttcactgata gatacaagag ccataagaac
     5101 ctcagatcct tccgtattta gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga
     5161 gccatgagaa cgaaccattg agatcatact tactttgcat gtcactcaaa aattttgcct
     5221 caaaactggt gagctgaatt tttgcagtta aagcatcgtg tagtgttttt cttagtccgt
     5281 tatgtaggta ggaatctgat gtaatggttg ttggtatttt gtcaccattc atttttatct
     5341 ggttgttctc aagttcggtt acgagatcca tttgtctatc tagttcaact tggaaaatca
     5401 acgtatcagt cgggcggcct cgcttatcaa ccaccaattt catattgctg taagtgttta
     5461 aatctttact tattggtttc aaaacccatt ggttaagcct tttaaactca tggtagttat
     5521 tttcaagcat taacatgaac ttaaattcat caaggctaat ctctatattt gccttgtgag
     5581 ttttcttttg tgttagttct tttaataacc actcataaat cctcatagag tatttgtttt
     5641 caaaagactt aacatgttcc agattatatt ttatgaattt ttttaactgg aaaagataag
     5701 gcaatatctc ttcactaaaa actaattcta atttttcgct tgagaacttg gcatagtttg
     5761 tccactggaa aatctcaaag cctttaacca aaggattcct gatttccaca gttctcgtca
     5821 tcagctctct ggttgcttta gctaata
//
