LOCUS       sequence-258217-        3446 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Single spacer array with spacer #3 for FnCas12a..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3446)
  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 3446)
  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-3
FEATURES             Location/Qualifiers
     source          1..3446
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..35
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     primer_bind     complement(219..236)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      389..475
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      567..594
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        613..704
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(705..773,774..1565)
                     /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(923..942)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1607..2062
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1694..1713)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1904..1925
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2173..2761
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2662..2681
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2915..2932
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2947..3087
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3013..3032)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3173..3195
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3333..3352
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ttgacagcta gctcagtcct aggtataatg ctagcgctag cgaattcgag ctcggtacca
       61 agacgtcaac tcgaccctgt ctaagaactt taaataattt ctactgttgt agatctgtac
      121 ataaccttcg ggcatggcac aacggtctaa gaactttaaa taatttctac tgttgtagat
      181 aagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa
      241 cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct
      301 gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc
      361 catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
      421 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
      481 gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc
      541 ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt
      601 tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
      661 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
      721 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
      781 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
      841 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
      901 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc
      961 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     1021 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     1081 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     1141 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     1201 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     1261 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     1321 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     1381 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     1441 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     1501 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     1561 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttacgc gccctgtagc
     1621 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     1681 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     1741 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     1801 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     1861 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     1921 actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg
     1981 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     2041 aaaatattaa cgtttacaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     2101 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2161 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2221 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2281 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     2341 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2401 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2461 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2521 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2581 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2641 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2701 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2761 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2821 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     2881 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     2941 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
     3001 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     3061 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     3121 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgacggt ctccgggagc
     3181 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagaa ggagatggcg
     3241 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     3301 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     3361 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatctgctca
     3421 tgtttgacag cttatcatcg atgcat
//
