LOCUS       sequence-258219-        3446 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Single spacer array with spacer #2 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-2
FEATURES             Location/Qualifiers
     source          1..3446
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..27
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     165..184
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        279..313
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     primer_bind     complement(497..514)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      667..753
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      845..872
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        891..982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(983..1051,1052..1843)
                     /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(1201..1220)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1885..2340
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1972..1991)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2182..2203
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2451..3039
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2940..2959
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3193..3210
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3225..3365
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3291..3310)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
       61 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
      121 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
      181 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
      241 aggatctgct catgtttgac agcttatcat cgatgcattt gacagctagc tcagtcctag
      301 gtataatgct agcgctagcg aattcgagct cggtaccaag acgtcaactc gaccctgtct
      361 aagaacttta aataatttct actgttgtag atcgtatgtt gcatcacctt caccctctaa
      421 cggtctaaga actttaaata atttctactg ttgtagataa gcttggctgt tttggcggat
      481 gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac
      541 agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag
      601 tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
      661 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
      721 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
      781 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt
      841 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt
      901 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
      961 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     1021 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     1081 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     1141 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     1201 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     1261 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     1321 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     1381 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     1441 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     1501 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     1561 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     1621 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     1681 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     1741 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     1801 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     1861 ctcatatata ctttagattg atttacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     1921 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     1981 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2041 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2101 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2161 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2221 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2281 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     2341 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2401 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2461 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2521 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2581 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     2641 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2701 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2761 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2821 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2881 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2941 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3001 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3061 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3121 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3181 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     3241 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc
     3301 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     3361 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3421 catccgctta cagacaagct gtgacg
//
