LOCUS       sequence-227117-        7486 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses RPCC7116RT-Cas1-Cas2 under pT7lac promoter, encodes 
            RPCC7116CRISPR array 1 RC, compatible with SENECA acquisition 
            readout.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7486)
  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 7486)
  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_0386_pET30-RT-CRISPR_Rivularia_sp._PCC_7116_ARRAY_1_RC
FEATURES             Location/Qualifiers
     source          1..7486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(409..1224)
                     /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(409..1224)
                     /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     1132..1151
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1216..1235
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1346..1934
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1835..1854
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2088..2105
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2120..2259
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2185..2204)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2345..2367
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2361..2552)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3361..4443)
                     /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     4405..4424
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4444..4521)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4727..4746
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4834..4853
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4834..4852
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4853..4877
                     /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             4892..4914
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(7399..7417)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7413..7460
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ccaggtattt gcgtcttagg acaagttatg cttccattgt tagaatttgt tgacaatcga
       61 ttaacgcaac aataagggtt tcagccctta aattacttgg ttatgcttcc aggggctgac
      121 agcttttggg tatagtgaca gaacctgatt acgaaggttt ccaactaatc cgaattaacc
      181 caatcggtgg gggtcccgcg gccgctattc ttttgattta taagggattt tgccgatttc
      241 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
      301 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
      361 ttatttttct aaatacattc aaatatgtat ccgctcatga attaattctt agaaaaactc
      421 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
      481 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
      541 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
      601 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
      661 gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc
      721 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
      781 acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
      841 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
      901 ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
      961 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
     1021 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     1081 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     1141 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga
     1201 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag
     1261 ttttattgtt catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     1321 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     1381 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     1441 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     1501 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     1561 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     1621 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     1681 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     1741 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     1801 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     1861 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     1921 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggtc ttttgctggc
     1981 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2041 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2101 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     2161 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     2221 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     2281 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     2341 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     2401 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     2461 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     2521 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     2581 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     2641 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     2701 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     2761 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     2821 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagatcat tcatgttgtt
     2881 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     2941 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     3001 acgatcatgc gcacccgtgg ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa
     3061 cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc
     3121 gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc
     3181 cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagatagt cataagtgcg
     3241 gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     3301 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     3361 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3421 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     3481 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     3541 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     3601 acatgagcta tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag
     3661 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     3721 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     3781 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     3841 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     3901 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccat cttcatggga
     3961 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     4021 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     4081 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     4141 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     4201 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     4261 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     4321 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     4381 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     4441 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     4501 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     4561 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     4621 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     4681 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     4741 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     4801 agaggatcga gatcgatctc gatcccgcga aattaatacg actcactata ggggaattgt
     4861 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatataca
     4921 tatgggatac gagaacatca gataccttca caagccgtac ttcgcagaaa cgattactga
     4981 gaagccacca cagactgagg aagatgactc accattccag aagaacatct ggaacgctga
     5041 gatggctgca atctatctta tcgagcaggg tacaaacatc tacaaagact accagcgttt
     5101 cataatacac gttagtgaga agccaaaact tgaggtccct atcagggacg tacaacagat
     5161 catcgtcttt ggaaatatcc agctaagcac gcctgttatc caggcctgcc tgaaggagca
     5221 gatcccagtt gttttcctgt ctcagaccgg aacataccat ggccatctgt ggtcggagaa
     5281 gtcaatccac cttgataacc agttggttca ggctgagcga aggaacgacg acttattcag
     5341 attctcagtt agccgagcag tggttctagg gaagcttctg aactctaagc aactactgat
     5401 gagattcaac agaaggagaa agattggtaa ggtcgaagag gcaatctacg gtatcaacca
     5461 ggacatagac gcattgaatt acgtggacaa cctggacacg ctgaggggat acgagggtat
     5521 cgcagccgcc cgttacttcc ccgccttcgg taacttgata acaaacccaa agttcagctt
     5581 cagtcaaagg ttcaggcagc caccaactga cgaaattaac tctctactgt ccttcggtta
     5641 cacattgtta ttcaacaacg ttttatcctt catcatcact gagggcttgt cgccgtacat
     5701 aggacacttc cattacggtg acaagcagaa aacttacctc gcattcgacc ttatggaaga
     5761 gttcagatcg cctatcgtag actctcttgt attgaagata attaacaaga gcctattcaa
     5821 gccccaagac tttgacgttg tcgcctctac cggcggtgtt tatttgtctc agaccagccg
     5881 tagggtgttc ctgaagcaat tcgagaacag aatgaacgag gaaatctccc acccggacct
     5941 gatctcccag gttacttacc gtcatgctat ccagttgcag gtcagacgat acaaaaggtg
     6001 cctgctgagt gacaacatct acgagtcttt cctaagggct gactaattaa ctttaagaag
     6061 gagatataca tatgatgata tcatctcagt tcatagacat aaacaacttc cagagagcat
     6121 gggagaaagt cgcagacaag agagggtgtg ctggagtaga cggagagaca atctcctcat
     6181 tcgccagtaa ccagactgtg aacgtctacc aactcatgaa tagtgtggca gacggatctt
     6241 accagccgtt cccatgtaaa caagtcatca tcccaaagag gaatgggtcc cagagggaac
     6301 taaagatacc gaccatcagg gataggatag tgcagcaggc tctcctaaac gtcatctcac
     6361 ctcttatgga agaaaagttc tctccagtct ctttcgcata caggccaaac ttgagctaca
     6421 ttaacgccgt ggagaagatc gcagattgga gagatatggg gtacgtttgg gtcctggacg
     6481 ccgacattgt gaaattcttc gacaacatcg atcaccacag acttttgcag caagtgcgac
     6541 tacacatcga tcaccctggc atcctgtgcc tgataaaggc atggatctct gtcggcgtcg
     6601 agacacgaga gggtttgatt cttccacaga aaggaatccc acagggcgct gtcatctcac
     6661 ctatcttagc taacatctac ctgcacgagt tcgacgagat aatctctgca tctgacctag
     6721 agatagtaag atacgccgac gacttcttag tactatccac cagccaggag agaatagcca
     6781 tagccaagtc ccaggttata gacttgctcg actcattggg tctagagatt aacacagaca
     6841 agacacagat cacgtctttc gagagaggat tcagattcct tggacacggg tttctgtccg
     6901 acgcaatatt tccagtggac accaacaagg ctaaactgaa aagtggaatc gagacaaaca
     6961 gagaaaagac taggacccga aagacgagca agaagaagct ctaccataat ccttacagaa
     7021 ataagaaggt cgtatgacgt acgttaactt taagaaggag aacttaagat gcttgtggtg
     7081 gtggtttatg acattcccaa tgacaagagg cgaacaaaac tgtctaactt cttggaaggg
     7141 tacggacagc gtgtgcagtt ctcagttttc gagtgcttct taaacctgga cgaaatgaga
     7201 cagctattcg agaaaagcaa gaagattgta aagccgtctg aggataacgt gagattctac
     7261 tggatatcag aggacgcagt gtctcgtgtg cttaccatag gctctgagca ccccaatgcc
     7321 ccacctaatt attacgtcat ctaactcgag atccggctgc taacaaagcc cgaaaggaag
     7381 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     7441 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggata
//
