LOCUS       sequence-252330-        3579 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Three-spacer array 4 for FnCas12a..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3579)
  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 3579)
  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-3/1/2
FEATURES             Location/Qualifiers
     source          1..3579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..27
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     166..185
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        280..314
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     primer_bind     complement(630..647)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      800..886
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      978..1005
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1024..1115
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1116..1184,1185..1976)
                     /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(1334..1353)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2018..2473
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2105..2124)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2584..3172
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3073..3092
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3326..3343
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3358..3498
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3424..3443)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
       61 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
      121 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
      181 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
      241 gaggatctgc tcatgtttga cagcttatca tcgatgcatt tgacagctag ctcagtccta
      301 ggtataatgc tagcgctagc gaattcgagc tcggtaccaa gacgtcaact cgaccctgtc
      361 taagaacttt aaataatttc tactgttgta gatctgtaca taaccttcgg gcatggcacg
      421 ctggtctaag aactttaaat aatttctact gttgtagatc tttacacttt atgcttccgg
      481 ctcgtgagtg tctaagaact ttaaataatt tctactgttg tagatcgtat gttgcatcac
      541 cttcaccctc taacggtcta agaactttaa ataatttcta ctgttgtaga taagcttggc
      601 tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc
      661 ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
      721 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga
      781 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
      841 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga
      901 tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc
      961 caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac
     1021 tcttttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     1081 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     1141 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     1201 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     1261 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     1321 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     1381 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     1441 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     1501 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     1561 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     1621 aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc
     1681 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     1741 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     1801 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     1861 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     1921 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     1981 cagaccaagt ttactcatat atactttaga ttgatttacg cgccctgtag cggcgcatta
     2041 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2101 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2161 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2221 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2281 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aacttgaaca
     2341 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
     2401 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     2461 acgtttacaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2521 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2581 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2641 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2701 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     2761 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2821 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2881 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2941 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3001 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3061 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3121 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3181 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3241 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3301 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     3361 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     3421 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     3481 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3541 tgtctgctcc cggcatccgc ttacagacaa gctgtgacg
//
