LOCUS       sequence-252341-        3443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Single spacer array with spacer #1 for FnCas12a..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3443)
  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 3443)
  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
FEATURES             Location/Qualifiers
     source          1..3443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        33..67
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     primer_bind     complement(247..264)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      417..503
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      595..622
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        641..732
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(733..801,802..1593)
                     /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(951..970)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1635..2090
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1722..1741)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2201..2789
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2690..2709
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2943..2960
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2975..3115
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3041..3060)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3201..3223
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3362..3381
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tgctcatgtt tgacagctta tcatcgatgc atttgacagc tagctcagtc ctaggtataa
       61 tgctagcgct agcgaattcg agctcggtac caagacgtca actcgagtct aagaacttta
      121 aataatttct actgttgtag atctttacac tttatgcttc cggctcgatc gagtctaaga
      181 actttaaata atttctactg ttgtagataa gcttggctgt tttggcggat gagagaagat
      241 tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc
      301 tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg
      361 tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa
      421 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
      481 acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc
      541 ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg
      601 ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt tttctaaata
      661 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
      721 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
      781 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
      841 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
      901 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
      961 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     1021 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     1081 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     1141 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     1201 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     1261 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     1321 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     1381 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     1441 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     1501 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     1561 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     1621 ctttagattg atttacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     1681 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     1741 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     1801 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     1861 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     1921 gttctttaat agtggactct tgttccaaac ttgaacaaca ctcaacccta tctcgggcta
     1981 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     2041 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt aaaaggatct
     2101 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     2161 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2221 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2281 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2341 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2401 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2461 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2521 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2581 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2641 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2701 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2761 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2821 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2881 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     2941 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     3001 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
     3061 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     3121 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     3181 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     3241 gaaacgcgcg aggcagcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     3301 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     3361 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     3421 cgatgcgtcc ggcgtagagg atc
//
