LOCUS       sequence-388293-        6949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  eLbuCas13a 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: pBA615
FEATURES             Location/Qualifiers
     source          1..6949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(3..20)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(36..65)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(144..178)
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     terminator      187..230
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     rep_origin      571..1116
                     /direction=RIGHT
                     /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     1017..1036
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1233..1250
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             1489..2148
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1555..1574)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2219..2253
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     RBS             5916..5927
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        5945..6000
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    5946..5964
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             6016..6642
                     /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     complement(6092..6111)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      complement(6680..6726)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     6879..6896
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
ORIGIN
        1 caatgatgat gatgatgatg gtcgacggcg ctattcagat cctcttctga gatgagtttt
       61 tgttcgggcc caagcattgg cgagtagcga agacgttatc aaagagttgt tttagtcccc
      121 ttcatttttg gggtggtcta aatactagta ttatacctag gactgagcta gctgtcaaga
      181 attccagaaa tcatccttag cgaaagctaa ggattttttt tatctgaaat cgttgtagct
      241 gtgttgacat ctggtttggc agcggaccaa aacgaaaaaa ggcccccctt tcgggaggcc
      301 tcttttctgg aatttggtac cgagggatgt ttaaaccgtc tgcaattatc ggccagttca
      361 gtctagatgc gaattcaggg ttgagaagag aaaagaaaac cgccgatcct gtccaccgca
      421 ttactgcaag gtagtggaca agaccggcgg tcttaagttt tttggctgaa acaacttata
      481 tcgtatgggg ctgacttcag gtgctacatt tgaagagata aattgcactg aaatctagaa
      541 atattttatc tgattaataa gatgatcttc ttgagatcgt tttggtctgc gcgtaatctc
      601 ttgctctgaa aacgaaaaaa ccgccttgca gggcggtttt tcgaaggttc tctgagctac
      661 caactctttg aaccgaggta actggcttgg aggagcgcag tcaccaaaac ttgtcctttc
      721 agtttagcct taaccggcgc atgacttcaa gactaactcc tctaaatcaa ttaccagtgg
      781 ctgctgccag tggtgctttt gcatgtcttt ccgggttgga ctcaagacga tagttaccgg
      841 ataaggcgca gcggtcggac tgaacggggg gttcgtgcat acagtccagc ttggagcgaa
      901 ctgcctaccc ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca taacagcgga
      961 atgacaccgg taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc gccaggggga
     1021 aacgcctggt atctttatag tcctgtcggg tttcgccacc actgatttga gcgtcagatt
     1081 tcgtgatgct tgtcaggggg gcggagccta tggaaaaacg gctttgccgc ggccctctca
     1141 cttccctgtt aagtatcttc ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca
     1201 tttccgctcg ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag gaagcggaat
     1261 atatcctgta tcacatattc tgctgacgca ccggtgcagc cttttttctc ctgccacatg
     1321 aagcacttca ctgacaccct catcagtgcc aacatagtaa gccagtatac actccgctag
     1381 cgcttgatcg ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg
     1441 ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa
     1501 atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca
     1561 tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt
     1621 ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc
     1681 cgcctgatga atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata
     1741 tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg
     1801 ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg
     1861 gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc
     1921 gtttcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac
     1981 aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg
     2041 atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg
     2101 cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tgatatcgag
     2161 ctcgcttgga ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt
     2221 cagaacgctc ggttgccgcc gggcgttttt tattggtgga aacacagaaa aaagcccgca
     2281 cctgacagtg cgggcttttt ttttcgacca aaggtgtact gcagtgcact agtggttcgt
     2341 aacatctctg taactgctca gcggaccaaa acgaaaaaag acgcttttca gcgtcttatt
     2401 gttcgtcttt ggtaccgagg gatttattag ttttcagact ttttctcttc cattttgtat
     2461 tcaaacataa tcttcaccaa ttttgaaagt tcctcggaat tgcggtcagt cattaacttc
     2521 ttcttcttaa gattcttaag atgcacgatt ttttcgctct ccagtgtctg aataccgatc
     2581 ttcttgtccg ccccgatttt gaatgtggct acaaaaccat actccttaag gatatcaact
     2641 actgatttca ttacggcatt tttcagtttg cgatcgtagg acagcaattt acgcaaattt
     2701 tcaaggactt ccagcagtga gatctcggcg tgaggaatat aattgaagtg ggcgatgtaa
     2761 tttcgaatat aaaggtcctt cttttcttgc ttcaacacct tgatattcgc ggaactgtac
     2821 ttgttgatct taacttcatc attttgatgt aattccttgt agaatttaat gtatttctct
     2881 acaatctgac cccctttgta cttcacgttc ttcttatttt caaagttgaa gatctcttcg
     2941 atgtattggt tttctgggaa ctcaccttta aggcggaagc gcagatcgcg ttcccaaatt
     3001 gaggtatatc cgactaaacg atgtaaaatg cgcagcaaca ggccctgcag taaattcagc
     3061 tcattgaatt ctaccttgtt cttaaggtgg gtgtactcct caatattctc aatagcttgc
     3121 ttgtaacttt catagtcctc atctgtaaac ttctcgtctt tacggggacg agcgtatttg
     3181 cggtgcagat tttcctgcat cttatggttc ttctcgatct cattcttttt attggagtat
     3241 ttcttcagct cttcgatcga gatcttgtat ccagccttgt cggcaatttt ctccagtaag
     3301 tttaacatgc catatttctt aatattgtag aaggcacggt gtttgataat gttctcgccg
     3361 tcaaagtaga tcttgtttgt atcgaacttt ttcagttcct tgttatcctt aactttgttg
     3421 ccattgaaat caagaaattt tccaatctcg tcggcctcta attcgaagtc ttccgtcacg
     3481 cggttgttgt caaggttaag caggttaatt aactccaatt ggtcagagaa cgcttcttcc
     3541 ttattggcag actgatattt ttcaagacta cccttcagat ttgtcagctc cttgtgattc
     3601 aaaagcttta agataaggta gaacatgttt aaacgctcag tatacttcgg gatgtttccc
     3661 agtttgatct cacgcaggaa ttcattgatc tcatagggga tcttgatatt attgttctgc
     3721 tcgtacttct tcaagaactt gtcaaactct tgcttctttt ctgctaatga cgtatttgtt
     3781 tcttcatccg agccgatgta aatcagactt aaacgaccgt tattagcaag atacgtcata
     3841 aatcccttta agaagatctt ttgaatgaaa tcaatataag tgtccttttc ttcttcgtcc
     3901 tgattaccgg cattaatcat gtacagggac tggatattcg ccaagtattc ctttggagtc
     3961 ttctcctgga tgtcttcaaa cttttgcagc ttgtaaaacc cagtctttaa gttgcgttta
     4021 tcgttcttgt tcaattcgat gatttcttta gaaatctcga agaaattacc attgttcgac
     4081 atgaaataat tcaggaactc cccgtaatag atgtttttaa gcaaatagat ttgggcatca
     4141 ataatctcct tagtcttatt gtcgtcgttt gttttcggag ttttccagta aatcccaaga
     4201 ctattcttca ggtcatcaat gcgcgaatat aatttggtga aagacgggac gaatgggata
     4261 tttttattga caaactcgaa gcgtgtacgt ttaaggtaat tcagaatttt atacttttcg
     4321 agatagcgga acacgttggc agagttcagt tgacgaaaga tcttcaattt cagttttttc
     4381 tcgttaatct cattctgaaa catcttcttt gagatttcgg atggcgcaat gttcttgaac
     4441 gcaaagatat ccttaccttc tgattccaag ttgaagtgga caataccgtg acggatggaa
     4501 gaaatcgcct cgtcgatgtt ggcgaagaaa tcttcgattt cattcttgtt gtccatattg
     4561 aaatcgtacg aatagaacat ttttaagttc tccttaacct tgttcttctt attttcatta
     4621 tagatcttat caacttctcc ggacacgtac ttctcttcac ctttattgtt cttaactgtt
     4681 ttgccgcgca tacgcccagt aatatcattc tcgttctccg tttcaagaat gttgcgaaga
     4741 gaaaagtagg ccacagatga caccccaatg atgttgcgaa gaaacgcttc attctgacgg
     4801 ttgcgggcga tgaaatctga agtggcaatt tcgccgtctt gcaaataata attatacttt
     4861 ccgctattac ggacgtacgt gtcaagtttg tttaacagct tattttcgat caattttttc
     4921 aagttctggt attcaaagat acgcttaatt ttgtcattcg agatattact taagcgctta
     4981 tatacgtagt tcttcagcaa ctgactcatt tcgatttcca cgaaatgcga aaacgcgtat
     5041 ttgatgttct tgtcgttcaa ctcttctttg tctaagtaat acttgtaaaa tacttggctc
     5101 tttttcaact cactgatgtc cggtaccttc tcgatcaact ctttcatgtt attaacattc
     5161 tggatttctt cgtagatgat ttttgcaaag ttttccttgt cattcttacg tccaataatt
     5221 tcgtggtaga actcgcgaat cttgtatttc tctgacttcg taaggttctc aatttcaaga
     5281 acaagttttg caatgtcctc ttccttgtaa agcttatcaa aggcttcttt cacattgctt
     5341 acataagcgt cacgtttcgc tgactcacga taataatcgt aaataatgtt acgctttgac
     5401 ttaccttcaa ccttctcgat gttattctca ttaatctttt gataattcgc cttattcttt
     5461 tcaaatgagt acttcaggct gttgattttg ttcagtttct tcttaatgtc gttacgaaaa
     5521 acttccaatt cctccgagtt cacgttttca ttcagataga tctttttcaa cacggcgaag
     5581 tttttcttgt cacggacatc gctctcaagg atgtcagtct cagaatactc acgatcaatg
     5641 ttctcctttt tcccattctt caaactcaag gtattgtctt taagatagac cattttgttt
     5701 gagaagaatt tctttaattt cccaatgcgt ttttgatttt ccttggtttc cgtgctgctg
     5761 ggattcttga tatacatgtc aaggcgcata ttaagcaacg ccgacagacg ttcgtctgtg
     5821 cgattttcgt ccgattctga cttcactaag cggccttcgg acgtgtactt cttatgcgaa
     5881 atgcctccta ccttcgtcac tttcatgtaa gatcctttct cctctttaga tcttttgaat
     5941 tcttttctct atcactgata gggagtggta aaataactct atcaacgata gagtgtcaac
     6001 aaaaattagg aattaatgat gtctagatta gataaaagta aagtgattaa cagcgcatta
     6061 gagctgctta atgaggtcgg aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta
     6121 ggtgtagagc agcctacatt gtattggcat gtaaaaaata agcgggcttt gctcgacgcc
     6181 ttagccattg agatgttaga taggcaccat actcactttt gccctttaga aggggaaagc
     6241 tggcaagatt ttttacgtaa taacgctaaa agttttagat gtgctttact aagtcatcgc
     6301 gatggagcaa aagtacattt aggtacacgg cctacagaaa aacagtatga aactctcgaa
     6361 aatcaattag cctttttatg ccaacaaggt ttttcactag agaatgcatt atatgcactc
     6421 agcgctgtgg ggcattttac tttaggttgc gtattggaag atcaagagca tcaagtcgct
     6481 aaagaagaaa gggaaacacc tactactgat agtatgccgc cattattacg acaagctatc
     6541 gaattatttg atcaccaagg tgcagagcca gccttcttat tcggccttga attgatcata
     6601 tgcggattag aaaaacaact taaatgtgaa agtgggtctt aagacgtcga tatctggcga
     6661 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct ttcgactgag cctttcgttt
     6721 tatttgatgc ctggcagttg cctactctcg catgggcaga ccccacacta ccatcggcgc
     6781 tacggcgttt cacttctgag ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc
     6841 caggcaaatt ctgttttatc agaccgcttc tgcgttctga tttaatctgt atcaggctga
     6901 aaatcttctc tcatccgcca aaacacccaa gctggagagc gtttaaact
//
