LOCUS       sequence-369170-        6949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LbuCas13a Negative Control (RFP targeting).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6949)
  AUTHORS   Adler BA, Hessler T, Cress BF, Lahiri A, Mutalik VK, Barrangou R, 
            Banfield J, Doudna JA
  TITLE     Broad-spectrum CRISPR-Cas13a enables efficient phage genome editing.
  JOURNAL   Nat Microbiol. 2022 Dec;7(12):1967-1979. doi: 
            10.1038/s41564-022-01258-x. Epub 2022 Oct 31.
  PUBMED    36316451
REFERENCE   2  (bases 1 to 6949)
  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"; doi: 
            "10.1038/s41564-022-01258-x"; journalName: "Nat Microbiol"; date: 
            "2022-12"; volume: "7"; issue: "12"; pages: "1967-1979"
COMMENT     Sequence Label: pBA620
FEATURES             Location/Qualifiers
     source          1..6949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(138..181)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        190..224
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     CDS             303..332
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             348..365
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(421..438)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      591..637
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(675..1301)
                     /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     1206..1225
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        1317..1372
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    1353..1371
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1390..1401
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      complement(5064..5098)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(5169..5828)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5743..5762
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(6067..6084)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6201..6746)
                     /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(6281..6300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctagactga actggccgat aattgcagac ggtttaaaca tccctcggta ccaaattcca
       61 gaaaagaggc ctcccgaaag gggggccttt tttcgttttg gtccgctgcc aaaccagatg
      121 tcaacacagc tacaacgatt tcagataaaa aaaatcctta gctttcgcta aggatgattt
      181 ctggaattct tgacagctag ctcagtccta ggtataatac tagtatttag accaccccaa
      241 aaatgaaggg gactaaaaca actctttgat aacgtcttcg ctactcgcca atgcttgggc
      301 ccgaacaaaa actcatctca gaagaggatc tgaatagcgc cgtcgaccat catcatcatc
      361 atcattgagt ttaaacgctc tccagcttgg gtgttttggc ggatgagaga agattttcag
      421 cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg
      481 cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc
      541 cgatggtagt gtggggtctg cccatgcgag agtaggcaac tgccaggcat caaataaaac
      601 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaactcgc
      661 cagatatcga cgtcttaaga cccactttca catttaagtt gtttttctaa tccgcatatg
      721 atcaattcaa ggccgaataa gaaggctggc tctgcacctt ggtgatcaaa taattcgata
      781 gcttgtcgta ataatggcgg catactatca gtagtaggtg tttccctttc ttctttagcg
      841 acttgatgct cttgatcttc caatacgcaa cctaaagtaa aatgccccac agcgctgagt
      901 gcatataatg cattctctag tgaaaaacct tgttggcata aaaaggctaa ttgattttcg
      961 agagtttcat actgtttttc tgtaggccgt gtacctaaat gtacttttgc tccatcgcga
     1021 tgacttagta aagcacatct aaaactttta gcgttattac gtaaaaaatc ttgccagctt
     1081 tccccttcta aagggcaaaa gtgagtatgg tgcctatcta acatctcaat ggctaaggcg
     1141 tcgagcaaag cccgcttatt ttttacatgc caatacaatg taggctgctc tacacctagc
     1201 ttctgggcga gtttacgggt tgttaaacct tcgattccga cctcattaag cagctctaat
     1261 gcgctgttaa tcactttact tttatctaat ctagacatca ttaattccta atttttgttg
     1321 acactctatc gttgatagag ttattttacc actccctatc agtgatagag aaaagaattc
     1381 aaaagatcta aagaggagaa aggatcttac atgaaagtga cgaaggtagg aggcatttcg
     1441 cataagaagt acacgtccga aggccgctta gtgaagtcag aatcggaaga aaatcgcaca
     1501 gacgaacgtc tgtcggcgtt gcttaatatg cgccttgaca tgtatatcaa gaatcccagc
     1561 agcacggaaa ccaaggaaaa tcaaaaacgc attgggaaat taaagaaatt cttctcaaac
     1621 aaaatggtct atcttaaaga caataccttg agtttgaaga atgggaaaaa ggagaacatt
     1681 gatcgtgagt attctgagac tgacatcctt gagagcgatg tccgtgacaa gaaaaacttc
     1741 gccgtgttga aaaagatcta tctgaatgaa aacgtgaact cggaggaatt ggaagttttt
     1801 cgtaacgaca ttaagaagaa actgaacaaa atcaacagcc tgaagtactc atttgaaaag
     1861 aataaggcga attatcaaaa gattaatgag aataacatcg agaaggttga aggtaagtca
     1921 aagcgtaaca ttatttacga ttattatcgt gagtcagcga aacgtgacgc ttatgtaagc
     1981 aatgtgaaag aagcctttga taagctttac aaggaagagg acattgcaaa acttgttctt
     2041 gaaattgaga accttacgaa gttagagaaa tacaagattc gcgagttcta ccacgaaatt
     2101 attggacgta agaatgacaa ggaaaacttt gcaaaaatca tctacgaaga aatccagaat
     2161 gttaataaca tgaaagagtt gatcgagaag gtaccggaca tgagtgagtt gaaaaagagc
     2221 caagtatttt acaagtatta cttagacaaa gaagagttga acgacaagaa catcaaatac
     2281 gcgttttgtc atttcgtgga aatcgaaatg agtcagttgc tgaagaacta cgtatataag
     2341 cgcttaagta atatctcgaa tgacaaaatt aagcgtatct ttgaatacca gaacttgaaa
     2401 aaattgatcg aaaataagct gttaaacaaa cttgacacgt acgtccgtaa ttgtggaaag
     2461 tataattatt atttgcaaga cggcgaaatt gccacttcag atttcatcgc ccgcaaccgt
     2521 cagaatgaag cgtttcttcg caacatcatt ggggtgtcat ctgtggccta cttttctctt
     2581 cgcaacattc ttgaaacgga gaacgagaat gatattactg ggcgtatgcg cggcaaaaca
     2641 gttaagaaca ataaaggtga agagaagtac gtgtccggag aagttgataa gatctataat
     2701 gaaaataaga agaacgaggt taaggagaac ttaaaaatgt tctattcgta cgatttcaat
     2761 atggacaaca agaatgaaat cgaagatttc ttcgccaaca tcgacgaggc gatttcttcc
     2821 atccgtcacg gtattgtcca cttcaacttg gaattagaag gtaaggatat ctttgcgttc
     2881 aagaacattg cgccatccga aatctcaaag aagatgtttc agaatgagat taacgagaaa
     2941 aaactgaaat tgaagatctt tcgtcaactg aactctgcca acgtgttccg ctatctcgaa
     3001 aagtataaaa ttctgaatta ccttaaacgt acacgcttcg agtttgtcaa taaaaatatc
     3061 ccattcgtcc cgtctttcac caaattatat tcgcgcattg atgacctgaa gaatagtctt
     3121 gggatttact ggaaaactcc gaaaacaaac gacgacaata agactaagga gattattgat
     3181 gcccaaatct atttgcttaa aaacatctat tacggggagt tcctgaatta tttcatgtcg
     3241 aacaatggta atttctttga gatttctaaa gaaatcatcg aattgaacaa gaacgataaa
     3301 cgcaacttaa agactgggtt ttacaagctg caaaagtttg aagacatcca ggagaagatt
     3361 ccaaaggaat acttggcgaa tatccagtcc ctgtacatga ttaatgccgg taatcaggac
     3421 gaagaagaaa aggacactta tattgatttc attcaaaaga tcttcttaaa gggatttatg
     3481 acgtatcttg ctaataacgg tcgtttaagt ctgatttaca tcggctcgga tgaagaaaca
     3541 aatacgtcat tagcagaaaa gaagcaagag tttgacaagt tcttgaagaa gtacgagcag
     3601 aacaataata tcaagatccc ctatgagatc aatgaattcc tgcgtgagat caaactggga
     3661 aacatcctga agtatactga gcgtttaaac atgttctacc ttatcttaaa gcttttgaat
     3721 cacaaggagc tgacaaatct gaagggtagt cttgaaaaat atcagtctgc caataaggaa
     3781 gaagcgttct ctgaccaatt ggagttaatt aacctgctta accttgacaa caaccgcgtg
     3841 acggaagact tcgaattaga ggccgacgag attggaaaat ttcttgattt caatggcaac
     3901 aaagttaagg ataacaagga actgaaaaag ttcgatacaa acaagatcta ctttgacggc
     3961 gagaacatta tcaaacaccg tgccttctac aatattaaga aatatggcat gttaaactta
     4021 ctggagaaaa ttgccgacaa ggctggatac aagatctcga tcgaagagct gaagaaatac
     4081 tccaataaaa agaatgagat cgagaagaac cataagatgc aggaaaatct gcaccgcaaa
     4141 tacgctcgtc cccgtaaaga cgagaagttt acagatgagg actatgaaag ttacaagcaa
     4201 gctattgaga atattgagga gtacacccac cttaagaaca aggtagaatt caatgagctg
     4261 aatttactgc agggcctgtt gctgcgcatt ttacatcgtt tagtcggata tacctcaatt
     4321 tgggaacgcg atctgcgctt ccgccttaaa ggtgagttcc cagaaaacca atacatcgaa
     4381 gagatcttca actttgaaaa taagaagaac gtgaagtaca aagggggtca gattgtagag
     4441 aaatacatta aattctacaa ggaattacat caaaatgatg aagttaagat caacaagtac
     4501 agttccgcga atatcaaggt gttgaagcaa gaaaagaagg acctttatat tcgcaattac
     4561 atcgcccact tcaattatat tcctcacgcc gagatctcac tgctggaagt ccttgaaaat
     4621 ttgcgtaaat tgctgtccta cgatcgcaaa ctgaaaaatg ccgtaatgaa atcagtagtt
     4681 gatatcctta aggagtatgg ttttgtagcc acattcaaaa tcggggcgga caagaagatc
     4741 ggtattcaga cactggagag cgaaaaaatc gtgcatctta agaatcttaa gaagaagaag
     4801 ttaatgactg accgcaattc cgaggaactt tgcaaattgg tgaagattat gtttgaatac
     4861 aaaatggaag agaaaaagtc tgaaaactaa taaatccctc ggtaccaaag acgaacaata
     4921 agacgctgaa aagcgtcttt tttcgttttg gtccgctgag cagttacaga gatgttacga
     4981 accactagtg cactgcagta cacctttggt cgaaaaaaaa agcccgcact gtcaggtgcg
     5041 ggcttttttc tgtgtttcca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca
     5101 aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg
     5161 atatcaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt
     5221 ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc
     5281 accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc
     5341 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgaaacgaaa
     5401 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca
     5461 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
     5521 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
     5581 accagctcac cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca
     5641 agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag
     5701 gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc
     5761 tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt
     5821 ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt
     5881 agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgcccgat caagcgctag
     5941 cggagtgtat actggcttac tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg
     6001 tggcaggaga aaaaaggctg caccggtgcg tcagcagaat atgtgataca ggatatattc
     6061 cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc
     6121 ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag
     6181 agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc
     6241 tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6301 cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg
     6361 ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg
     6421 ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg
     6481 taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac
     6541 tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa
     6601 ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag
     6661 ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga
     6721 ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata aaatatttct
     6781 agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata
     6841 agttgtttca gccaaaaaac ttaagaccgc cggtcttgtc cactaccttg cagtaatgcg
     6901 gtggacagga tcggcggttt tcttttctct tctcaaccct gaattcgca
//
