LOCUS       sequence-220285-        7061 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MrRT-Cas1-Cas2 under pT7lac promoter, encodes MrCRISPR 
            array 1, compatible with SENECA acquisition readout.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7061)
  AUTHORS   Schmidt F, Cherepkova MY, Platt RJ
  TITLE     Transcriptional recording by CRISPR spacer acquisition from RNA.
  JOURNAL   Nature. 2018 Oct;562(7727):380-385. doi: 10.1038/s41586-018-0569-1. 
            Epub 2018 Oct 3.
  PUBMED    30283135
REFERENCE   2  (bases 1 to 7061)
  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/s41586-018-0569-1"; journalName: "Nature"; date: "2018-10";
            volume: "562"; issue: "7727"; pages: "380-385"
COMMENT     Sequence Label: 
            pFS_0357_pET30-RT-CRISPR_Micromonospora_rosaria_ARRAY_1
FEATURES             Location/Qualifiers
     source          1..7061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(411..1226)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(411..1226)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1134..1153
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1218..1237
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1348..1936
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1837..1856
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2090..2107
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2122..2261
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2187..2206)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2347..2369
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2363..2554)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3363..4445)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4407..4426
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4446..4523)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4729..4748
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4836..4855
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4836..4854
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4855..4879
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             4894..4916
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(6974..6992)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6988..7035
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ccaggtagca cagatgcgcg caaacctctt ctcgcaagtc aagccagcat ctcccaacgc
       61 ctgcgacctg gaccacaccc gccacagcac ctcttcgcca cggtccgcgc gacgatctcc
      121 atcccagcac gtcagtccca ccgattttcg ggtagcatcg ccatccacag cccaacccag
      181 cgggcataga aacgtcccgc ggccgctatt cttttgattt ataagggatt ttgccgattt
      241 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
      301 tattaacgct tacaatttca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
      361 gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac
      421 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
      481 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
      541 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
      601 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
      661 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
      721 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
      781 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
      841 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
      901 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
      961 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     1021 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     1081 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     1141 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa
     1201 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     1261 agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     1321 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1381 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1441 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     1501 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1561 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1621 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1681 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1741 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1801 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1861 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1921 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg tcttttgctg
     1981 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     2041 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     2101 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     2161 ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     2221 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
     2281 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     2341 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     2401 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     2461 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     2521 ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc
     2581 tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg
     2641 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc
     2701 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag
     2761 gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg
     2821 gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagatc attcatgttg
     2881 ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg
     2941 attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga
     3001 gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga
     3061 aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata
     3121 ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga
     3181 cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagata gtcataagtg
     3241 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     3301 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
     3361 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3421 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
     3481 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
     3541 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
     3601 taacatgagc tatcttcggt atcgtcgtat cccactaccg agatgtccgc accaacgcgc
     3661 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
     3721 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
     3781 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
     3841 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
     3901 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc atcttcatgg
     3961 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
     4021 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
     4081 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
     4141 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
     4201 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
     4261 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
     4321 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
     4381 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
     4441 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
     4501 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat
     4561 taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc
     4621 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
     4681 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     4741 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     4801 gtagaggatc gagatcgatc tcgatcccgc gaaattaata cgactcacta taggggaatt
     4861 gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata
     4921 catatgacca gggtgtctat ccctaaaccc gatggcggaa tacgtagctt ggctatagga
     4981 gctattgaag atagaattgt cgaaagagct gttctggatg ttcttgatcc agttgtggac
     5041 cccacacttt ccccgtggag ttttgcatac aggagaggct taggcgtgag ggatgcagtt
     5101 cgagctttgg cagaagctag ggaatctggt ctagcttttg ttgttcgttg tgatattgat
     5161 gattgttttg attcgatccc gagatggcct ctacttagga gactaagaga attggtttct
     5221 gacgctgaat tggttgcctt ggtagaaaga ttggttggca gaccagtcac tggagaaagg
     5281 gcttccggtg gtaggggatt gcatcagggc ggatcgctca gtcctttgtt ggccaaccta
     5341 tatttggata cctttgatcg agcattgatg cgtcatggac accgtgtagt tagatatgga
     5401 gatgatatag ctatatctgt tcccgatagg ccaactggac ttagagtatt ggatttggct
     5461 gatgcagaag cagaagccct gtctttgaga ctaaatactg atgatagaca agtcatagct
     5521 tttgatgaag gagtcccttt ctgtggtcag gttgttactg catcaagcgg gccaacagct
     5581 gatttacaag ctaaacctct gcaaggtaca gtctttgtta caacccaagg agcattgcta
     5641 agagtaaaag gagaacgact tcgtgtcgaa gatggagaca ggctactggc taatgtaaat
     5701 ttgaagcgtg tgaggcaaat agtctgtttt ggtagagtag gagtgacttc taccttgtta
     5761 caaagaattg tagaacgagg aatagaactt gcttggttat atgaggatgg tagacatgca
     5821 gctagagtaa gtgggttgga tggtactgat ccagaggtcc gtttagccca atatagagct
     5881 gctgatgatg cacgtcaggc actgagaata gctaggcagt tagtcgctgg taaagtaaca
     5941 aatatgagag ttggtctttt gcgagctgct agggctcaac aagctccaga gcttgctgat
     6001 cgtcaggcac gattggccac agctcgacag tcagctttga ttgcagattc tactgctgaa
     6061 ttgatgggtt atgagggttc agctactaga gattattttg ctggtctttc acaaattttg
     6121 ggtcctgagt ggggctttac tactcgtcaa agaaggcctc ctcctgaccc cgtgaatgcc
     6181 atgcttagtt ttggatatac cttgctgact aatgaggcac tcacggcctg tcagctggct
     6241 ggacttgatc catatctcgg tatgttacat tctcctagaa gaaatagacc ttctttggca
     6301 ttggatttaa tcgaagaact ccgacccgtt gttgttgatg ctaccgttat taggctagta
     6361 cgaacaggac aagttactcc taagaacttt actctgacag atgatagagg atgtagattg
     6421 gatgatcatg gtagacgtgc ctttctggat gcttatgagc gtagaatgct aacacttgtc
     6481 catcatcctg tagaacaaag aagaataccg tggagacatg ttttgttagc acaagctaga
     6541 acacttgctg ctgtattgtc atcgcgtcgt cccgaatata gacccgtagt atggcgttaa
     6601 cgtacgttaa ctttaagaag gagaacttaa gatgtcccgt gtttgttatg tagttgccta
     6661 tgatatagcc gatgacgacc gaagggccga tctcgctatg tttctttcag ggtacggtcc
     6721 gcgtgttcag ttgtctgtgt ttgaagtcga attgccggat gctgatacag ctgttgcttt
     6781 tagggagcgt ttgcgtagtt taatagaccc taacgacgat caagttcgtc tgtatcgact
     6841 cactccacag gcattgggac agagaattat ttacggaaga aggaccattg aggagagagt
     6901 tgacttttgg atcgtgtaac tcgagatccg gctgctaaca aagcccgaaa ggaagctgag
     6961 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     7021 ttgaggggtt ttttgctgaa aggaggaact atatccggat a
//
