LOCUS       sequence-381844-        3039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Consensus I-C CRISPR Leader and a CRISPR locus with two consensus 
            I-C repeats, for integration assays..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3039)
  AUTHORS   Santiago-Frangos A, Henriques WS, Wiegand T, Gauvin CC, Buyukyoruk 
            M, Graham AB, Wilkinson RA, Triem L, Neselu K, Eng ET, Lander GC, 
            Wiedenheft B
  TITLE     Structure reveals why genome folding is necessary for site-specific 
            integration of foreign DNA into CRISPR arrays.
  JOURNAL   Nat Struct Mol Biol. 2023 Nov;30(11):1675-1685. doi: 
            10.1038/s41594-023-01097-2. Epub 2023 Sep 14.
  PUBMED    37710013
REFERENCE   2  (bases 1 to 3039)
  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/s41594-023-01097-2"; journalName: "Nat Struct Mol Biol"; 
            date: "2023-11"; volume: "30"; issue: "11"; pages: "1675-1685"
COMMENT     Sequence Label: pCon-ICl_Con-ICr
FEATURES             Location/Qualifiers
     source          1..3039
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(818..834)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(818..834)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(831..853)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    842..858
                     /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)."
     promoter        complement(join(866..872,873..890,891..896))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    911..932
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1049..1066)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1220..1808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1300..1319)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1979..2770,2771..2839))
                     /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     2602..2621
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2840..2944)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3012..3030
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
      421 tgcatctaga tccatgggag tgcgaacgtg tagcgaccgg gattttacgg ggaggttcgc
      481 ggatttgcac ggcgttgaat ttgttaggga tttttttaag cacggtggga atttgaggac
      541 ggaggcgtgt ttgggccaca gaagagcaga ggttcgcgga attgggcggt ttttgtctgg
      601 cagtacagtg ggttaggtta ggtgggtcgc gcccttcacg ggcgcgtgga ttgaaacatc
      661 agccggacgt tgtagtagtc gagcgcggtg tcgcgccctt cacgggcgcg tggattgaaa
      721 cgccgggtgg tgacttctgg acggcgttcg tcaaggatcc atcggatccc gggcccgtcg
      781 actgcagagg cctgcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
      841 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
      901 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
      961 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1021 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1081 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1141 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1201 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1261 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1321 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     1381 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     1441 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     1501 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     1561 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     1621 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     1681 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     1741 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     1801 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     1861 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     1921 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     1981 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     2041 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     2101 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     2161 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     2221 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     2281 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     2341 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     2401 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     2461 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     2521 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     2581 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     2641 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     2701 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     2761 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     2821 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     2881 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     2941 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     3001 taacctataa aaataggcgt atcacgaggc cctttcgtc
//
