LOCUS       sequence-490799-        4031 bp    DNA     circular SYN 27-OCT-2025
DEFINITION  Bacterial expression of Panoptes locus with 300bp of context 
            upstream and 150bp downstream of the ECOR31 operon cloned under the 
            pTet promoter; p15a ori.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4031)
  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 4031)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 27, 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: pBA1729
FEATURES             Location/Qualifiers
     source          1..4031
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(7..633)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     538..557
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        649..704
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    685..703
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      2196..2267
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2283..2310
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2338..2355)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2472..3017)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2552..2571)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3131..3225)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3246..3905)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3820..3839
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3906..4008)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 gacgtcttaa gacccacttt cacatttaag ttgtttttct aatccgcata tgatcaattc
       61 aaggccgaat aagaaggctg gctctgcacc ttggtgatca aataattcga tagcttgtcg
      121 taataatggc ggcatactat cagtagtagg tgtttccctt tcttctttag cgacttgatg
      181 ctcttgatct tccaatacgc aacctaaagt aaaatgcccc acagcgctga gtgcatataa
      241 tgcattctct agtgaaaaac cttgttggca taaaaaggct aattgatttt cgagagtttc
      301 atactgtttt tctgtaggcc gtgtacctaa atgtactttt gctccatcgc gatgacttag
      361 taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc tttccccttc
      421 taaagggcaa aagtgagtat ggtgcctatc taacatctca atggctaagg cgtcgagcaa
      481 agcccgctta ttttttacat gccaatacaa tgtaggctgc tctacaccta gcttctgggc
      541 gagtttacgg gttgttaaac cttcgattcc gacctcatta agcagctcta atgcgctgtt
      601 aatcacttta cttttatcta atctagacat cattaattcc taatttttgt tgacactcta
      661 tcgttgatag agttatttta ccactcccta tcagtgatag agaaaagaat tcaaaagatc
      721 ttttaagaag gagatataca ttgagaaatc tttagtgtaa agatttaata attatccttt
      781 aagtgtagat tacaagcaga acaattgaat taatcgccac attgaaaata taaaacctca
      841 acacacttga tgtatttaac aagacatagg attaaaccca ttaaaattaa aaaaatttaa
      901 aatacaaacc tcgttttggc cgggtataaa tacaatttaa accgcaacgt ttttaagtaa
      961 ataaattcat ttgcaactaa gcattcaggg ttgatataat ataaccactt agataataat
     1021 ccatgttagt ttggagttaa tatgtacata gcgatagatg gcgatgatgt ggggagaaaa
     1081 ataacctcat catacttatc caactcagaa gagaggttga catacataag caacaaatta
     1141 aatgatacca caaaaaaaat atctaaaatg cttttatcca atggatttga gataattttc
     1201 caagccgcag atggtgttac agcgaaaaca gacaacgagg tcaatcttaa ctttgttttt
     1261 gataagataa aatcctattc ctttgacgaa attacatttt cagctggtgt gggagcgaat
     1321 ctaagagaag cctatgttgc actgttaaat tctaaaagca acggtaagaa tatgatttct
     1381 atttacaaag acatattata atggtgcgaa atgtttagaa taataagatt tgactggttg
     1441 ttaaaaacgg tatcactaat ctctttgttt gtcacaatac tttcattttg gatctttaag
     1501 acagttctgt cttctgactt aagcctgctc aaattgttat caatatcatc attagcatcc
     1561 attatcataa acttctttct cctttcatca aagatatcaa gactaatatg gggaacaata
     1621 aggtttttca agaaagacat atatcctgac ttaaatggaa tttgggttgg acatataatc
     1681 acagaaacaa atgatgaatt tgaagtccgc gcaaaaataa agcaagcact ccttgtaaca
     1741 aagatagaaa tgcacggccc cacagttaaa tctgttacgc tagaagcaac gccaactaag
     1801 gaactggata acaacaaatt gtattatgta tataaatcaa cccctaaaaa ccctagttgg
     1861 agcgagtata ttggctcgac tatattcgac gtaattgaaa gtaacaatgc gctccaactt
     1921 tccggtcgat attatacaga cagaaaatcc gtggggagaa tatcaataaa aagaatatca
     1981 cttaacactg actccgatat atctttttat taactttcgg aatgtttaat taccacaatc
     2041 agccatgttc gttcttttaa aatagaagga taattcgcgc tactgatata ttaatcatgg
     2101 aaaaactcct ttcagtagcc attttcccta gaaaaatacc cctgcatacc tcatatactc
     2161 catggatcca aactcgagta aggatctcca ggcatcaaat aaaacgaaag gctcagtcga
     2221 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca
     2281 ctggctcacc ttcgggtggg cctttctgcg tttataccta gggatatatt ccgcttcctc
     2341 gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg
     2401 gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg
     2461 gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca
     2521 aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc
     2581 tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg
     2641 ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct
     2701 ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg
     2761 tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg
     2821 atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt
     2881 ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa
     2941 ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca
     3001 gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc tagatttcag
     3061 tgcaatttat ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgttac
     3121 tagtgcttgg attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat
     3181 ccagatggag ttctgaggtc attactggat ctatcaacag gagtccaagc gagctcgata
     3241 tcaaattacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg
     3301 ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc
     3361 ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata
     3421 ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac
     3481 atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct
     3541 tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa
     3601 aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc
     3661 agctcaccgt ctttcattgc catacgaaat tccggatgag cattcatcag gcgggcaaga
     3721 atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc
     3781 gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca
     3841 aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc
     3901 tccattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt
     3961 gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaac gtctcatttt
     4021 cgccagatat c
//
