LOCUS       sequence-373572-        6944 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTet-LbuCas13a crRNA entry vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6944)
  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 6944)
  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: pBA559
FEATURES             Location/Qualifiers
     source          1..6944
                     /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             298..327
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             343..360
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(416..433)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      586..632
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(670..1296)
                     /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     1201..1220
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        1312..1367
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    1348..1366
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1385..1396
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      complement(5059..5093)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(5164..5823)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5738..5757
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(6062..6079)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6196..6741)
                     /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(6276..6295)
                     /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 gagacctcgt ttacctatcg gtctcatgct tgggcccgaa
      301 caaaaactca tctcagaaga ggatctgaat agcgccgtcg accatcatca tcatcatcat
      361 tgagtttaaa cgctctccag cttgggtgtt ttggcggatg agagaagatt ttcagcctga
      421 tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta
      481 gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg
      541 gtagtgtggg gtctgcccat gcgagagtag gcaactgcca ggcatcaaat aaaacgaaag
      601 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa ctcgccagat
      661 atcgacgtct taagacccac tttcacattt aagttgtttt tctaatccgc atatgatcaa
      721 ttcaaggccg aataagaagg ctggctctgc accttggtga tcaaataatt cgatagcttg
      781 tcgtaataat ggcggcatac tatcagtagt aggtgtttcc ctttcttctt tagcgacttg
      841 atgctcttga tcttccaata cgcaacctaa agtaaaatgc cccacagcgc tgagtgcata
      901 taatgcattc tctagtgaaa aaccttgttg gcataaaaag gctaattgat tttcgagagt
      961 ttcatactgt ttttctgtag gccgtgtacc taaatgtact tttgctccat cgcgatgact
     1021 tagtaaagca catctaaaac ttttagcgtt attacgtaaa aaatcttgcc agctttcccc
     1081 ttctaaaggg caaaagtgag tatggtgcct atctaacatc tcaatggcta aggcgtcgag
     1141 caaagcccgc ttatttttta catgccaata caatgtaggc tgctctacac ctagcttctg
     1201 ggcgagttta cgggttgtta aaccttcgat tccgacctca ttaagcagct ctaatgcgct
     1261 gttaatcact ttacttttat ctaatctaga catcattaat tcctaatttt tgttgacact
     1321 ctatcgttga tagagttatt ttaccactcc ctatcagtga tagagaaaag aattcaaaag
     1381 atctaaagag gagaaaggat cttacatgaa agtgacgaag gtaggaggca tttcgcataa
     1441 gaagtacacg tccgaaggcc gcttagtgaa gtcagaatcg gaagaaaatc gcacagacga
     1501 acgtctgtcg gcgttgctta atatgcgcct tgacatgtat atcaagaatc ccagcagcac
     1561 ggaaaccaag gaaaatcaaa aacgcattgg gaaattaaag aaattcttct caaacaaaat
     1621 ggtctatctt aaagacaata ccttgagttt gaagaatggg aaaaaggaga acattgatcg
     1681 tgagtattct gagactgaca tccttgagag cgatgtccgt gacaagaaaa acttcgccgt
     1741 gttgaaaaag atctatctga atgaaaacgt gaactcggag gaattggaag tttttcgtaa
     1801 cgacattaag aagaaactga acaaaatcaa cagcctgaag tactcatttg aaaagaataa
     1861 ggcgaattat caaaagatta atgagaataa catcgagaag gttgaaggta agtcaaagcg
     1921 taacattatt tacgattatt atcgtgagtc agcgaaacgt gacgcttatg taagcaatgt
     1981 gaaagaagcc tttgataagc tttacaagga agaggacatt gcaaaacttg ttcttgaaat
     2041 tgagaacctt acgaagttag agaaatacaa gattcgcgag ttctaccacg aaattattgg
     2101 acgtaagaat gacaaggaaa actttgcaaa aatcatctac gaagaaatcc agaatgttaa
     2161 taacatgaaa gagttgatcg agaaggtacc ggacatgagt gagttgaaaa agagccaagt
     2221 attttacaag tattacttag acaaagaaga gttgaacgac aagaacatca aatacgcgtt
     2281 ttgtcatttc gtggaaatcg aaatgagtca gttgctgaag aactacgtat ataagcgctt
     2341 aagtaatatc tcgaatgaca aaattaagcg tatctttgaa taccagaact tgaaaaaatt
     2401 gatcgaaaat aagctgttaa acaaacttga cacgtacgtc cgtaattgtg gaaagtataa
     2461 ttattatttg caagacggcg aaattgccac ttcagatttc atcgcccgca accgtcagaa
     2521 tgaagcgttt cttcgcaaca tcattggggt gtcatctgtg gcctactttt ctcttcgcaa
     2581 cattcttgaa acggagaacg agaatgatat tactgggcgt atgcgcggca aaacagttaa
     2641 gaacaataaa ggtgaagaga agtacgtgtc cggagaagtt gataagatct ataatgaaaa
     2701 taagaagaac gaggttaagg agaacttaaa aatgttctat tcgtacgatt tcaatatgga
     2761 caacaagaat gaaatcgaag atttcttcgc caacatcgac gaggcgattt cttccatccg
     2821 tcacggtatt gtccacttca acttggaatt agaaggtaag gatatctttg cgttcaagaa
     2881 cattgcgcca tccgaaatct caaagaagat gtttcagaat gagattaacg agaaaaaact
     2941 gaaattgaag atctttcgtc aactgaactc tgccaacgtg ttccgctatc tcgaaaagta
     3001 taaaattctg aattacctta aacgtacacg cttcgagttt gtcaataaaa atatcccatt
     3061 cgtcccgtct ttcaccaaat tatattcgcg cattgatgac ctgaagaata gtcttgggat
     3121 ttactggaaa actccgaaaa caaacgacga caataagact aaggagatta ttgatgccca
     3181 aatctatttg cttaaaaaca tctattacgg ggagttcctg aattatttca tgtcgaacaa
     3241 tggtaatttc tttgagattt ctaaagaaat catcgaattg aacaagaacg ataaacgcaa
     3301 cttaaagact gggttttaca agctgcaaaa gtttgaagac atccaggaga agattccaaa
     3361 ggaatacttg gcgaatatcc agtccctgta catgattaat gccggtaatc aggacgaaga
     3421 agaaaaggac acttatattg atttcattca aaagatcttc ttaaagggat ttatgacgta
     3481 tcttgctaat aacggtcgtt taagtctgat ttacatcggc tcggatgaag aaacaaatac
     3541 gtcattagca gaaaagaagc aagagtttga caagttcttg aagaagtacg agcagaacaa
     3601 taatatcaag atcccctatg agatcaatga attcctgcgt gagatcaaac tgggaaacat
     3661 cctgaagtat actgagcgtt taaacatgtt ctaccttatc ttaaagcttt tgaatcacaa
     3721 ggagctgaca aatctgaagg gtagtcttga aaaatatcag tctgccaata aggaagaagc
     3781 gttctctgac caattggagt taattaacct gcttaacctt gacaacaacc gcgtgacgga
     3841 agacttcgaa ttagaggccg acgagattgg aaaatttctt gatttcaatg gcaacaaagt
     3901 taaggataac aaggaactga aaaagttcga tacaaacaag atctactttg acggcgagaa
     3961 cattatcaaa caccgtgcct tctacaatat taagaaatat ggcatgttaa acttactgga
     4021 gaaaattgcc gacaaggctg gatacaagat ctcgatcgaa gagctgaaga aatactccaa
     4081 taaaaagaat gagatcgaga agaaccataa gatgcaggaa aatctgcacc gcaaatacgc
     4141 tcgtccccgt aaagacgaga agtttacaga tgaggactat gaaagttaca agcaagctat
     4201 tgagaatatt gaggagtaca cccaccttaa gaacaaggta gaattcaatg agctgaattt
     4261 actgcagggc ctgttgctgc gcattttaca tcgtttagtc ggatatacct caatttggga
     4321 acgcgatctg cgcttccgcc ttaaaggtga gttcccagaa aaccaataca tcgaagagat
     4381 cttcaacttt gaaaataaga agaacgtgaa gtacaaaggg ggtcagattg tagagaaata
     4441 cattaaattc tacaaggaat tacatcaaaa tgatgaagtt aagatcaaca agtacagttc
     4501 cgcgaatatc aaggtgttga agcaagaaaa gaaggacctt tatattcgca attacatcgc
     4561 ccacttcaat tatattcctc acgccgagat ctcactgctg gaagtccttg aaaatttgcg
     4621 taaattgctg tcctacgatc gcaaactgaa aaatgccgta atgaaatcag tagttgatat
     4681 ccttaaggag tatggttttg tagccacatt caaaatcggg gcggacaaga agatcggtat
     4741 tcagacactg gagagcgaaa aaatcgtgca tcttaagaat cttaagaaga agaagttaat
     4801 gactgaccgc aattccgagg aactttgcaa attggtgaag attatgtttg aatacaaaat
     4861 ggaagagaaa aagtctgaaa actaataaat ccctcggtac caaagacgaa caataagacg
     4921 ctgaaaagcg tcttttttcg ttttggtccg ctgagcagtt acagagatgt tacgaaccac
     4981 tagtgcactg cagtacacct ttggtcgaaa aaaaaagccc gcactgtcag gtgcgggctt
     5041 ttttctgtgt ttccaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc
     5101 agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagcga gctcgatatc
     5161 aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc
     5221 gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt
     5281 gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt
     5341 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaaa cgaaaaacat
     5401 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg
     5461 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa
     5521 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag
     5581 ctcaccgtct ttcattgcca tacgaaattc cggatgagca ttcatcaggc gggcaagaat
     5641 gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt
     5701 aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa
     5761 atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc
     5821 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     5881 tcttatttca ttatggtgaa agttggaacc tcttacgtgc ccgatcaagc gctagcggag
     5941 tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct tcatgtggca
     6001 ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata tattccgctt
     6061 cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg
     6121 aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa gtgagagggc
     6181 cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg aaatctgacg
     6241 ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6301 cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca ttccgctgtt
     6361 atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca gttcgctcca
     6421 agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta tccggtaact
     6481 atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca gccactggta
     6541 attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca
     6601 agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt ggtagctcag
     6661 agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca agagattacg
     6721 cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata tttctagatt
     6781 tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg
     6841 tttcagccaa aaaacttaag accgccggtc ttgtccacta ccttgcagta atgcggtgga
     6901 caggatcggc ggttttcttt tctcttctca accctgaatt cgca
//
