LOCUS       sequence-258212-        3442 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 3442)
  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 3442)
  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..3442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        36..70
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     primer_bind     complement(250..267)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      420..506
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      598..625
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        644..735
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(736..804,805..1596)
                     /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(954..973)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1638..2093
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1725..1744)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1935..1956
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2204..2792
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2693..2712
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2946..2963
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2978..3118
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3044..3063)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3204..3226
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3364..3383
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atctgctcat gtttgacagc ttatcatcga tgcatttgac agctagctca gtcctaggta
       61 taatgctagc gctagcgaat tcgagctcgg taccaagacg tcaactcgag tctaagaact
      121 ttaaataatt tctactgttg tagatcttta cactttatgc ttccggctcg atcgagtcta
      181 agaactttaa ataatttcta ctgttgtaga taagcttggc tgttttggcg gatgagagaa
      241 gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt
      301 gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg
      361 ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc
      421 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
      481 tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac
      541 ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga
      601 aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttttgttt atttttctaa
      661 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
      721 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
      781 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
      841 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
      901 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
      961 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     1021 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     1081 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     1141 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     1201 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     1261 cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa
     1321 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     1381 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     1441 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     1501 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     1561 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     1621 atactttaga ttgatttacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     1681 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     1741 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     1801 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     1861 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     1921 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg
     1981 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     2041 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa tttaaaagga
     2101 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     2161 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2221 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2281 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2341 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     2401 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2461 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     2521 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     2581 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     2641 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     2701 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     2761 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     2821 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     2881 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     2941 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     3001 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     3061 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     3121 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3181 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3241 accgaaacgc gcgaggcaga aggagatggc gcccaacagt cccccggcca cggggcctgc
     3301 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     3361 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     3421 cacgatgcgt ccggcgtaga gg
//
