LOCUS       sequence-150464-        3792 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Cas2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3792)
  AUTHORS   Shipman SL, Nivala J, Macklis JD, Church GM
  TITLE     Molecular recordings by directed CRISPR spacer acquisition.
  JOURNAL   Science. 2016 Jun 9. pii: aaf1175.
  PUBMED    27284167
REFERENCE   2  (bases 1 to 3792)
  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"; journalName: "Science. 
            2016 Jun 9. pii: aaf1175."
COMMENT     Sequence Label: pWUR Cas2
FEATURES             Location/Qualifiers
     source          1..3792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     CDS             394..438
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(476..494)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      490..537
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(708..1499)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(1500..1591)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1639..2377)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(2587..3669)
                     /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     3631..3650
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3670..3747)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg agtatgttgg tcgtggtcac tgaaaatgta
      121 cctccgcgct tacgaggcag attagccatc tggttgttgg aggtacgtgc aggggtatat
      181 gtaggtgatg tatccgcaaa aattcgtgaa atgatctggg aacaaatagc tggactggcg
      241 gaagaaggca atgtagtgat ggcatgggca acgaatacgg aaacgggatt tgagttccag
      301 acatttgggt taaacaggcg taccccggta gatttggatg gtttaaggtt ggtgtctttt
      361 ttacctgttt gagcggccgc actcgagtct ggtaaagaaa ccgctgctgc gaaatttgaa
      421 cgccagcaca tggactcgtc tactagcgca gcttaattaa cctaggctgc tgccaccgct
      481 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
      541 aaacctcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta
      601 aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtca
      661 tcgtggccgg atcttgcggc ccctcggctt gaacgaattg ttagacatta tttgccgact
      721 accttggtga tctcgccttt cacgtagtgg acaaattctt ccaactgatc tgcgcgcgag
      781 gccaagcgat cttcttcttg tccaagataa gcctgtctag cttcaagtat gacgggctga
      841 tactgggccg gcaggcgctc cattgcccag tcggcagcga catccttcgg cgcgattttg
      901 ccggttactg cgctgtacca aatgcgggac aacgtaagca ctacatttcg ctcatcgcca
      961 gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat ttagcgcctc aaatagatcc
     1021 tgttcaggaa ccggatcaaa gagttcctcc gccgctggac ctaccaaggc aacgctatgt
     1081 tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga tcgtggctgg ctcgaagata
     1141 cctgcaagaa tgtcattgcg ctgccattct ccaaattgca gttcgcgctt agctggataa
     1201 cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg gagaatctcg
     1261 ctctctccag gggaagccga agtttccaaa aggtcgttga tcaaagctcg ccgcgttgtt
     1321 tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg cttcaggccg
     1381 ccatccactg cggagccgta caaatgtacg gccagcaacg tcggttcgag atggcgctcg
     1441 atgacgccaa ctacctctga tagttgagtc gatacttcgg cgatcaccgc ttccctcata
     1501 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     1561 atatttgaat gtatttagaa aaataaacaa atagctagct cactcggtcg ctacgctccg
     1621 ggcgtgagac tgcggcgggc gctgcggaca catacaaagt tacccacaga ttccgtggat
     1681 aagcagggga ctaacatgtg aggcaaaaca gcagggccgc gccggtggcg tttttccata
     1741 ggctccgccc tcctgccaga gttcacataa acagacgctt ttccggtgca tctgtgggag
     1801 ccgtgaggct caaccatgaa tctgacagta cgggcgaaac ccgacaggac ttaaagatcc
     1861 ccaccgtttc cggcgggtcg ctccctcttg cgctctcctg ttccgaccct gccgtttacc
     1921 ggatacctgt tccgcctttc tcccttacgg gaagtgtggc gctttctcat agctcacaca
     1981 ctggtatctc ggctcggtgt aggtcgttcg ctccaagctg ggctgtaagc aagaactccc
     2041 cgttcagccc gactgctgcg ccttatccgg taactgttca cttgagtcca acccggaaaa
     2101 gcacggtaaa acgccactgg cagcagccat tggtaactgg gagttcgcag aggatttgtt
     2161 tagctaaaca cgcggttgct cttgaagtgt gcgccaaagt ccggctacac tggaaggaca
     2221 gatttggttg ctgtgctctg cgaaagccag ttaccacggt taagcagttc cccaactgac
     2281 ttaaccttcg atcaaaccac ctccccaggt ggttttttcg tttacagggc aaaagattac
     2341 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctactgaac cgctctagat
     2401 ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt
     2461 gtaattctca tgttagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
     2521 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
     2581 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2641 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
     2701 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
     2761 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg
     2821 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatgt ccgcaccaac
     2881 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
     2941 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
     3001 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
     3061 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
     3121 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
     3181 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
     3241 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
     3301 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
     3361 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
     3421 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
     3481 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
     3541 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
     3601 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
     3661 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
     3721 aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct
     3781 gcattaggaa at
//
