LOCUS       sequence-252327-        3579 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Three-spacer array 5 for FnCas12a..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3579)
  AUTHORS   Liao C, Ttofali F, Slotkowski RA, Denny SR, Cecil TD, Leenay RT, 
            Keung AJ, Beisel CL
  TITLE     Modular one-pot assembly of CRISPR arrays enables library generation
            and reveals factors influencing crRNA biogenesis.
  JOURNAL   Nat Commun. 2019 Jul 3;10(1):2948. doi: 10.1038/s41467-019-10747-3.
  PUBMED    31270316
REFERENCE   2  (bases 1 to 3579)
  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: "Nat 
            Commun."; date: "2019-07-3"; volume: "10(1):2948. doi"; pages: " 
            10.1038/s41467-019-10747-3"
COMMENT     Sequence Label: pcF-2/3/1
FEATURES             Location/Qualifiers
     source          1..3579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      197..785
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     686..705
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     939..956
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    971..1111
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1037..1056)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1197..1219
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1358..1377
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1472..1506
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     primer_bind     complement(1822..1839)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1992..2078
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2170..2197
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2216..2307
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2308..2376,2377..3168)
                     /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(2526..2545)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3210..3579,1..86)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3297..3316)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
       61 taacaaaata ttaacgttta caatttaaaa ggatctaggt gaagatcctt tttgataatc
      121 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
      181 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      241 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      301 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
      361 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      421 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      481 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      541 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      601 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      661 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      721 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      781 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      841 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      901 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      961 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     1021 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc
     1081 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
     1141 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga cggtctccgg
     1201 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agcaaggaga
     1261 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     1321 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     1381 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatct
     1441 gctcatgttt gacagcttat catcgatgca tttgacagct agctcagtcc taggtataat
     1501 gctagcgcta gcgaattcga gctcggtacc aagacgtcaa ctcgaccctg tctaagaact
     1561 ttaaataatt tctactgttg tagatcgtat gttgcatcac cttcaccctc tgctggtcta
     1621 agaactttaa ataatttcta ctgttgtaga tctgtacata accttcgggc atggcacgag
     1681 tgtctaagaa ctttaaataa tttctactgt tgtagatctt tacactttat gcttccggct
     1741 cgtaacggtc taagaacttt aaataatttc tactgttgta gataagcttg gctgttttgg
     1801 cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat
     1861 aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc
     1921 agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa
     1981 ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     2041 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     2101 ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc
     2161 aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt
     2221 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2281 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     2341 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     2401 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     2461 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     2521 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     2581 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     2641 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     2701 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     2761 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     2821 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     2881 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     2941 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3001 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     3061 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     3121 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     3181 gtttactcat atatacttta gattgattta cgcgccctgt agcggcgcat taagcgcggc
     3241 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3301 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3361 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3421 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3481 gacgttggag tccacgttct ttaatagtgg actcttgttc caaacttgaa caacactcaa
     3541 ccctatctcg ggctattctt ttgatttata agggatttt
//
