LOCUS       sequence-344928-        7285 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Encodes DisCas7-11 protein alone with a golden gate site for spacer 
            cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1978 to 2012)
  AUTHORS   Ozcan A, Krajeski R, Ioannidi E, Lee B, Gardner A, Makarova KS, 
            Koonin EV, Abudayyeh OO, Gootenberg JS
  TITLE     Programmable RNA targeting with the single-protein CRISPR effector 
            Cas7-11.
  JOURNAL   Nature. 2021 Sep 6. pii: 10.1038/s41586-021-03886-5. doi: 
            10.1038/s41586-021-03886-5.
  PUBMED    34489594
REFERENCE   2  (bases 1 to 7285)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2021 Sep 6. pii: 10.1038/s41586-021-03886-5. doi: 
            10.1038/s41586-021-03886-5."
COMMENT     Sequence Label: pDF0229 DisCas7-11 bacterial expression vector...
FEATURES             Location/Qualifiers
     source          1..7285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(108..767)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     682..701
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(768..870)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(1262..1279)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1396..1941)
                     /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(1476..1495)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1978..2012
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     RBS             2156..2167
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      6996..7066
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gaccgggtcg aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag
       61 gcgtagcaac caggcgttta agggcaccaa taactgcctt aaaaaaatta cgccccgccc
      121 tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca
      181 caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa
      241 tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca
      301 aaactggtga aactcaccca gggattggct gacacgaaaa acatattctc aataaaccct
      361 ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga
      421 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca
      481 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt
      541 gccatacgga attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga
      601 taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg
      661 gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc
      721 cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta
      781 gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
      841 tgaaagttgg aacctcttac gtgccgatca acgtgtcatt ttcgccaaaa gttggcccag
      901 ggcttcccgg tatcaacagg gacaccagga tttatttatt ctgcgaagtg atcttccgtc
      961 acaggtattt attcggcgca aagtgcgtcg ggtgatgctg ccaacttact gatttagtgt
     1021 atgatggtgt ttttgaggtg ctccagtggc ttctgtttct atcagctgtc cctcctgttc
     1081 agctactgac ggggtggtgc gtaacggcaa aagcaccgcc ggacatcagc gctagcggag
     1141 tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct tcatgtggca
     1201 ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata tattccgctt
     1261 cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg
     1321 aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa gtgagagggc
     1381 cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg aaatctgacg
     1441 ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1501 cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca ttccgctgtt
     1561 atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca gttcgctcca
     1621 agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta tccggtaact
     1681 atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca gccactggta
     1741 attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca
     1801 agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt ggtagctcag
     1861 agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca agagattacg
     1921 cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata tttctagttg
     1981 acggctagct cagtcctagg tacagtgcta gcggttggaa agccggtttt ctttgatgtc
     2041 acggaaccga gacctactgt tggtctcggg ttggaaagcc ggttttcttt gatgtcacgg
     2101 ttgccacccc ggtcttggac aggctgctca cctagtataa tgctagctac tagagaaaga
     2161 ggagaaatac tagatgacca ctaccatgaa gatcagcatc gagttcttgg agcccttccg
     2221 catgacgaag tggcaggagt ccacgcgccg caataagaac aataaagagt ttgtgcgtgg
     2281 ccaagcgttt gcccgttggc accgcaacaa gaaagataac accaagggtc gtccgtacat
     2341 tactggtacg ctcctgcgct cagcggtgat tcgctccgcg gagaacctgc tgacactgag
     2401 tgacggtaag atctccgaga aaacctgctg cccggggaaa ttcgacacgg aagataagga
     2461 ccgtttactc cagctgcgtc aacgctccac gctgcgttgg acagacaaga atccgtgtcc
     2521 cgacaatgcg gaaacgtatt gtccgttctg tgagctcctc ggccgctccg gaaatgacgg
     2581 taagaaagca gaaaagaagg actggcgctt ccgcatccat tttggtaacc tttccctgcc
     2641 gggcaagccc gacttcgacg gtccgaaggc aattggttca cagcgcgtat taaatcgtgt
     2701 ggattttaag tcgggaaagg cacatgactt cttcaaggct tatgaggtgg accatacccg
     2761 cttccctcgc tttgaagggg agatcaccat cgacaataaa gtgtccgccg aggcacgtaa
     2821 gttgctgtgt gacagcctta aattcacgga tcgcctgtgt ggagccctgt gcgtcatccg
     2881 cttcgacgag tatacaccgg ccgcggacag cggtaagcaa accgagaatg tgcaggcaga
     2941 gcccaacgca aacctggcgg agaaaactgc cgagcaaatt atctcgatcc tcgatgacaa
     3001 caagaaaacg gaatacaccc gccttttagc ggacgccatc cgctcattgc gtcgctcctc
     3061 aaaacttgtt gcaggtttgc ccaaggatca cgatggtaag gacgatcatt acttgtggga
     3121 cattgggaag aagaagaagg atgagaattc ggttaccatc cgtcagattc ttaccacgag
     3181 tgccgacacg aaggagctga agaacgctgg taagtggcgc gagttttgtg agaagcttgg
     3241 tgaggcgctt tatttgaaga gtaaggatat gagcggcggg ttgaagatta cacgccgtat
     3301 tctcggggac gcagagtttc atggcaagcc agatcgcttg gaaaagtccc gtagtgtctc
     3361 gatcggaagc gtgttaaagg aaaccgtggt gtgcggagaa ctcgtcgcaa agaccccgtt
     3421 cttctttggc gcgatcgacg aggatgcaaa acaaactgac ttacaggtac tgcttactcc
     3481 ggacaataag tatcgcctgc cgcgcagcgc cgtccgcggc atccttcgcc gcgatcttca
     3541 aacgtacttt gacagtccct gtaatgcgga gttaggcggt cgtccatgta tgtgcaagac
     3601 ctgtcgtatc atgcgtggca ttacggttat ggacgcgcgc tccgagtata acgcgccccc
     3661 ggagattcgt caccgtaccc gcattaatcc gtttacggga acagttgctg agggcgccct
     3721 gtttaatatg gaagtggccc cggaagggat cgtattccca ttccaattac gctatcgcgg
     3781 ctctgaggac gggctcccag acgcgctcaa gaccgtgctg aagtggtggg cagaggggca
     3841 agcttttatg tcaggggccg ccagtaccgg gaaaggccgt tttcgtatgg agaatgccaa
     3901 atatgaaacc ttggatctga gcgatgagaa tcaacgcaat gactacttga agaattgggg
     3961 ctggcgtgac gagaagggtc ttgaggagct taagaaacgt ttaaattcag gtctgccgga
     4021 gcctggcaat taccgtgacc cgaagtggca tgaaatcaac gtgtcaatcg agatggcgag
     4081 tcccttcatc aacggcgacc ctatccgcgc agctgtagac aagcgtggga cggacgtcgt
     4141 gaccttcgtt aagtacaagg cggaagggga agaagcaaag cccgtgtgcg catataaggc
     4201 ggagtcgttt cgcggcgtca ttcgttcggc tgtagcacgc attcacatgg aagatggagt
     4261 gcccctgact gagcttactc actcagattg cgagtgtttg ctctgtcaaa tcttcgggtc
     4321 ggaatatgag gctggtaaga ttcgtttcga agatttagta tttgagagtg acccggagcc
     4381 ggtcaccttc gatcacgttg caatcgatcg cttcaccggc ggtgccgctg acaagaagaa
     4441 gttcgacgac agtcctttgc cagggtcgcc ggcgcgccct ctgatgctga agggttcctt
     4501 ctggattcgc cgtgacgttc tcgaagatga ggagtattgc aaagcgctcg ggaaagcatt
     4561 agccgatgta aataatggtt tatatcctct gggtggaaaa tccgccatcg gatacggaca
     4621 agttaagagc ctgggtatca agggcgacga taagcgtatc agccgtttaa tgaatccggc
     4681 cttcgacgaa accgacgtgg cggttccaga gaagcctaaa accgatgccg aggtccgtat
     4741 cgaagctgag aaggtatatt atccccacta ctttgtggaa ccgcacaaga aggtagagcg
     4801 cgaggagaag ccgtgcggtc atcaaaagtt tcatgaggga cgccttaccg gtaagattcg
     4861 ttgtaaactt attacgaaaa cgcctctcat tgtgccggac accagcaatg atgacttctt
     4921 tcgccctgcg gacaaagagg cccgcaagga gaaggacgag tatcacaaat cctatgcctt
     4981 ctttcgcctg cataagcaaa tcatgatccc cggtagtgaa ctccgcggga tggtgtcctc
     5041 tgtttacgaa acggttacaa attcatgttt ccgtatcttt gacgagacta agcgtcttag
     5101 ttggcgcatg gatgccgatc accaaaacgt tttacaagat ttcttaccgg gccgtgttac
     5161 ggccgacggc aagcacattc aaaagttttc cgagactgca cgtgtaccgt tttatgacaa
     5221 aacgcaaaag cactttgaca ttctcgacga gcaggaaatt gccggtgaaa agccggtccg
     5281 catgtgggtg aagcgtttca ttaagcgcct gagcctggtg gaccctgcaa agcacccaca
     5341 aaagaagcaa gacaataagt ggaaacgccg caaggaaggt attgcgacgt tcattgagca
     5401 gaagaatggg tcttactatt ttaacgtggt caccaacaat ggctgcacca gctttcactt
     5461 gtggcacaag ccagacaact tcgaccaaga gaagttagag ggcatccaaa acggggagaa
     5521 gttagattgc tgggttcgtg atagccgtta ccaaaaggca tttcaagaaa tcccggagaa
     5581 cgatccggac ggatgggagt gtaaggaagg ttatctgcat gtggtgggcc cttcgaaggt
     5641 tgaatttagc gacaagaagg gcgatgtcat taataacttc caagggactc tcccatcagt
     5701 gcccaatgat tggaaaacga ttcgtacaaa cgactttaag aaccgcaaac gtaagaacga
     5761 gcccgttttc tgttgcgagg atgacaaggg gaactactac acgatggcca aatattgtga
     5821 aacgttcttc ttcgacctga aagagaacga ggagtatgag atccccgaga aggcacatat
     5881 caagtacaag gagttattac gcgtgtacaa caacaatccg caagcggttc ccgagagcgt
     5941 gttccagagt cgcgtagcgc gcgagaacgt agaaaagctg aagtctgggg atttggtgta
     6001 tttcaagcac aacgagaaat atgttgagga catcgtaccg gttcgtattt cacgtaccgt
     6061 agacgaccgc atgatcggaa agcgtatgtc ggctgatttg cgtccatgtc acggtgactg
     6121 ggtagaggac ggggacctct ctgcgttgaa tgcatatcca gagaagcgcc ttttattgcg
     6181 tcaccctaag ggcctttgcc cggcatgtcg ccttttcgga acgggtagct acaagggccg
     6241 cgttcgtttt ggcttcgcga gcctggagaa cgacccggag tggttgattc ctggaaagaa
     6301 tcccggtgat ccattccacg gcgggccagt gatgctgagc ttgctggagc gcccgcgtcc
     6361 cacctggagc attcctggca gtgataataa gttcaaagtt ccaggccgta agttctacgt
     6421 tcaccatcat gcttggaaaa cgattaagga cggtaaccat ccgaccaccg gcaaagccat
     6481 tgaacaatcg ccaaataacc gcacggttga agcgctcgcc ggtggcaaca gcttcagctt
     6541 cgaaattgcg ttcgaaaatt tgaaggagtg ggagttgggc ctgctgatcc atagcctgca
     6601 gcttgaaaag ggcttggccc ataagctcgg tatggctaag agcatgggct ttggctcagt
     6661 agagatcgac gttgagagtg tccgcctgcg taaagattgg aagcagtggc gcaatggtaa
     6721 ttcagagatc ccaaattggt tgggcaaggg gtttgcaaag ctgaaggagt ggttccgcga
     6781 cgagctggac ttcatcgaga atttgaagaa attactgtgg ttccccgaag gtgatcaagc
     6841 tccgcgcgta tgttacccca tgcttcgcaa gaaggatgac cctaatggaa actcaggtta
     6901 tgaggagttg aaagacggcg aatttaagaa ggaagatcgt cagaagaagc ttacaacacc
     6961 atggacgccg tgggcttgat actagagcca ggcatcaaat aaaacgaaag gtcagtcgaa
     7021 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact agagtcacac
     7081 tggctcacct ttcgggtggg cctttctgcg tttataccac gccttggacc ccttgggcct
     7141 gatcttctgt gccgcctgat actctccctt cgtaagatga ttcagcatga aaagttcatt
     7201 atttcagccc ggaagttcat tttcgggctg aaacctgttg gtttcctccg ataacccgct
     7261 tcagcgggtt tcaataactg agccg
//
