LOCUS       sequence-99597-e        3399 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive repeat-only E. coli CRISPR array.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3399)
  AUTHORS   Luo ML, Mullis AS, Leenay RT, Beisel CL
  TITLE     Repurposing endogenous type I CRISPR-Cas systems for programmable 
            gene repression.
  JOURNAL   Nucleic Acids Res. 2014 Oct 17. pii: gku971.
  PUBMED    25326321
REFERENCE   2  (bases 1 to 3399)
  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: "Nucleic 
            Acids Res. 2014 Oct 17. pii: gku971."
COMMENT     Sequence Label: pcrRNA.con
FEATURES             Location/Qualifiers
     source          1..3399
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        33..67
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     primer_bind     complement(203..220)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      373..459
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      551..578
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        597..688
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(689..757,758..1549)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(907..926)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1591..2046
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1678..1697)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2157..2745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2646..2665
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2899..2916
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2931..3071
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2997..3016)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3157..3179
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3318..3337
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tgctcatgtt tgacagctta tcatcgatgc atttgacagc tagctcagtc ctaggtataa
       61 tgctagcgct agcgaattcg agctcggtac cacctcgagt tccccgcgcc agcggggata
      121 aaccgaaaaa aaaaccccgc ccctgacagg gcggggtttt ttttaagctt ggctgttttg
      181 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga
      241 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact
      301 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga
      361 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
      421 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
      481 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat
      541 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg
      601 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      661 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      721 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      781 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      841 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      901 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
      961 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     1021 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     1081 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     1141 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     1201 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     1261 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1321 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     1381 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     1441 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1501 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1561 agtttactca tatatacttt agattgattt acgcgccctg tagcggcgca ttaagcgcgg
     1621 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     1681 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     1741 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     1801 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     1861 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaacttga acaacactca
     1921 accctatctc gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     1981 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     2041 caatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     2101 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2161 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2221 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2281 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     2341 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2401 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     2461 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2521 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2581 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2641 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2701 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2761 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2821 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2881 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     2941 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     3001 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     3061 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3121 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3181 tttcaccgtc atcaccgaaa cgcgcgaggc agcaaggaga tggcgcccaa cagtcccccg
     3241 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
     3301 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
     3361 ccggtgatgc cggccacgat gcgtccggcg tagaggatc
//
