LOCUS       sequence-150463-        4494 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Cas1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4494)
  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 4494)
  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: pWURA Cas1
FEATURES             Location/Qualifiers
     source          1..4494
                     /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             1027..1071
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1109..1127)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1123..1170
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(1341..2132,2133..2201))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1964..1983
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2202..2293)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2341..3079)
                     /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(3289..4371)
                     /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     4333..4352
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4372..4449)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg acctggcttc cccttaatcc cattccactc
      121 aaagatcgcg tctccatgat ctttctgcaa tatgggcaga tcgatgtaat agatggcgcg
      181 tttgtactta tcgacaagac agggatccgc actcatattc ctgttggctc ggttgcctgc
      241 atcatgctgg aacctggtac acgggtttcg catgcagctg tacgcctggc tgcgcaagtt
      301 ggaacattgt tggtatgggt gggggaagcg ggcgttcgtg tttatgcttc tggtcagcct
      361 ggaggtgcgc gttcagataa gctgctctat caggcaaaac ttgctctgga tgaagatttg
      421 cgtctgaagg tcgtacgtaa aatgtttgaa cttcggtttg gagaacctgc gcctgcccgg
      481 cgctccgtag agcaactcag aggtatagaa ggcagtcgcg tgcgggcaac ctacgcactt
      541 ctggcgaagc aatacggcgt gacatggaat ggacgtcgct acgatccgaa agactgggaa
      601 aagggcgata cgatcaacca atgcattagc gctgcaactt cctgtttata cggcgtaact
      661 gaagcggcga tacttgcagc tggttatgca ccagctattg ggtttgtgca tacaggaaag
      721 cctctttcct ttgtttacga tattgcagac atcattaaat ttgacactgt tgtaccgaaa
      781 gcttttgaga tagcgcgtcg taaccctggt gagccggacc gggaagtccg tttggcgtgc
      841 agggatattt ttcgcagtag taaaacatta gccaaattga ttccgcttat agaggacgtg
      901 cttgccgctg gagaaataca accgccggcc ccacctgaag atgcacagcc tgttgccatt
      961 ccgcttcctg tttcactggg agatgcaggc catcggagta gctgagcggc cgcactcgag
     1021 tctggtaaag aaaccgctgc tgcgaaattt gaacgccagc acatggactc gtctactagc
     1081 gcagcttaat taacctaggc tgctgccacc gctgagcaat aactagcata accccttggg
     1141 gcctctaaac gggtcttgag gggttttttg ctgaaacctc aggcatttga gaagcacacg
     1201 gtcacactgc ttccggtagt caataaaccg gtaaaccagc aatagacata agcggctatt
     1261 taacgaccct gccctgaacc gacgaccggg tcatcgtggc cggatcttgc ggcccctcgg
     1321 cttgaacgaa ttgttagaca ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     1381 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1441 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     1501 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1561 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1621 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1681 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1741 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1801 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1861 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1921 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1981 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     2041 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2101 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     2161 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     2221 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     2281 aaaaataaac aaatagctag ctcactcggt cgctacgctc cgggcgtgag actgcggcgg
     2341 gcgctgcgga cacatacaaa gttacccaca gattccgtgg ataagcaggg gactaacatg
     2401 tgaggcaaaa cagcagggcc gcgccggtgg cgtttttcca taggctccgc cctcctgcca
     2461 gagttcacat aaacagacgc ttttccggtg catctgtggg agccgtgagg ctcaaccatg
     2521 aatctgacag tacgggcgaa acccgacagg acttaaagat ccccaccgtt tccggcgggt
     2581 cgctccctct tgcgctctcc tgttccgacc ctgccgttta ccggatacct gttccgcctt
     2641 tctcccttac gggaagtgtg gcgctttctc atagctcaca cactggtatc tcggctcggt
     2701 gtaggtcgtt cgctccaagc tgggctgtaa gcaagaactc cccgttcagc ccgactgctg
     2761 cgccttatcc ggtaactgtt cacttgagtc caacccggaa aagcacggta aaacgccact
     2821 ggcagcagcc attggtaact gggagttcgc agaggatttg tttagctaaa cacgcggttg
     2881 ctcttgaagt gtgcgccaaa gtccggctac actggaagga cagatttggt tgctgtgctc
     2941 tgcgaaagcc agttaccacg gttaagcagt tccccaactg acttaacctt cgatcaaacc
     3001 acctccccag gtggtttttt cgtttacagg gcaaaagatt acgcgcagaa aaaaaggatc
     3061 tcaagaagat cctttgatct tttctactga accgctctag atttcagtgc aatttatctc
     3121 ttcaaatgta gcacctgaag tcagccccat acgatataag ttgtaattct catgttagtc
     3181 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     3241 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     3301 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3361 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     3421 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     3481 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     3541 ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt
     3601 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     3661 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     3721 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     3781 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     3841 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     3901 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     3961 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     4021 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     4081 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     4141 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     4201 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     4261 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     4321 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     4381 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     4441 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aaat
//
