LOCUS       sequence-258222-        3578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Three-spacer array 2 for FnCas12a..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3578)
  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 3578)
  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-1/3/2
FEATURES             Location/Qualifiers
     source          1..3578
                     /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(351..368)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      521..607
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      699..726
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        745..836
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(837..905,906..1697)
                     /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(1055..1074)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1739..2194
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1826..1845)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2036..2057
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2305..2893
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2794..2813
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3047..3064
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3079..3219
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3145..3164)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3305..3327
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3465..3484
                     /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 agatctttac
      121 actttatgct tccggctcgt gctggtctaa gaactttaaa taatttctac tgttgtagat
      181 ctgtacataa ccttcgggca tggcacgagt gtctaagaac tttaaataat ttctactgtt
      241 gtagatcgta tgttgcatca ccttcaccct ctaacggtct aagaacttta aataatttct
      301 actgttgtag ataagcttgg ctgttttggc ggatgagaga agattttcag cctgatacag
      361 attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg
      421 gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt
      481 gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca
      541 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
      601 gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc
      661 aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg
      721 cctttttgcg tttctacaaa ctcttttgtt tatttttcta aatacattca aatatgtatc
      781 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
      841 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
      901 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
      961 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     1021 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     1081 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     1141 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     1201 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     1261 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     1321 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     1381 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     1441 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     1501 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     1561 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     1621 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     1681 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttac
     1741 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1801 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1861 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1921 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     1981 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2041 ctcttgttcc aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa
     2101 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2161 gcgaatttta acaaaatatt aacgtttaca atttaaaagg atctaggtga agatcctttt
     2221 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2281 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2341 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2401 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     2461 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2521 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2581 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2641 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2701 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2761 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2821 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2881 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2941 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3001 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3061 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3121 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     3181 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     3241 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacg
     3301 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     3361 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     3421 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     3481 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     3541 aggatctgct catgtttgac agcttatcat cgatgcat
//
