LOCUS       sequence-379165-        2354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded plasmid with BsaI sites to clone sgRNA. The 6-nucleotide 
            barcode (sequence: CCTAGT) near the sgRNA insert region enables 
            demultiplexing replicates while sequencing the sgRNA region..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2354)
  AUTHORS   Otto RM, Turska-Nowak A, Brown PM, Reynolds KA
  TITLE     A continuous epistasis model for predicting growth rate given 
            combinatorial variation in gene expression and environment.
  JOURNAL   Cell Syst. 2024 Feb 21;15(2):134-148.e7. doi: 
            10.1016/j.cels.2024.01.003. Epub 2024 Feb 9.
  PUBMED    38340730
REFERENCE   2  (bases 1 to 2354)
  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"; doi: 
            "10.1016/j.cels.2024.01.003"; journalName: "Cell Syst"; date: 
            "2024-02-21- 21"; volume: "15"; issue: "2"; pages: "134-148.e7"
COMMENT     Sequence Label: pCRISPR3_BC6
FEATURES             Location/Qualifiers
     source          1..2354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(285..303)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             482..1276
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(536..555)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1146..1165
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      1307..1401
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1489..2077
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1978..1997
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2242..2328)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggaacccgac gaacgtgtat gtttcactac gtgtggcgtc ctggacaggg gatcgcttaa
       61 caccaactgg ttttcaggcg atgggtagcc gcagcacagt gaagacacgg ctatccatac
      121 tagatagaga acctccgata atgctgctac cgttttctaa tcttgacgat ctgtcggcca
      181 ttattagtac agcgaggcaa cactaggacg gatccccacc gagacctttg gtctcaacct
      241 ctcgaggtga agacgaaagg ggcatacata acatggagtc gtacgcctcg tgatacgcct
      301 atttttatag gttaatgtca tgataataat ggtttcttag acgtcggaat tgccagctgg
      361 ggcgccctct ggtaaggttg ggaagccctg caaagtaaac tggatggctt tcttgccgcc
      421 aaggatctga tggcgcaggg gatcaagatc tgatcaagag acaggatgag gatcgtttcg
      481 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
      541 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
      601 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
      661 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
      721 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
      781 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
      841 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
      901 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
      961 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     1021 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     1081 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     1141 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     1201 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     1261 tgacgagttc ttctgagcgg gactctgggg ttcgagagct cgcttggact cctgttgata
     1321 gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg
     1381 gcgtttttta ttggtgagaa tccaagcact agtcatgacc aaaatccctt aacgtgagtt
     1441 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     1501 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     1561 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     1621 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     1681 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     1741 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     1801 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     1861 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     1921 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     1981 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2041 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2101 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2161 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     2221 gccctaggtc tagggcggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     2281 gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctctagca
     2341 cgcgtaccac aatt
//
