LOCUS       sequence-379162-        2354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded plasmid with BsaI sites to clone sgRNA. The 6-nucleotide 
            barcode (sequence: GTATGA) 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_BC4
FEATURES             Location/Qualifiers
     source          1..2354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(281..299)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             478..1272
                     /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(532..551)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1142..1161
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      1303..1397
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1485..2073
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1974..1993
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2238..2324)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cccgacgaac gtgtatgttt cactacgtgt ggcgtcctgg acaggggatc gcttaacacc
       61 aactggtttt caggcgatgg gtagccgcag cacagtgaag acacggctat ccatactaga
      121 tagagaacct ccgataatgc tgctaccgtt ttctaatctt gacgatctgt cggccattat
      181 tagtacagcg aggcaactca tacacggatc cccaccgaga cctttggtct caacctctcg
      241 aggtgaagac gaaaggggca tacataacat ggagtcgtac gcctcgtgat acgcctattt
      301 ttataggtta atgtcatgat aataatggtt tcttagacgt cggaattgcc agctggggcg
      361 ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaagg
      421 atctgatggc gcaggggatc aagatctgat caagagacag gatgaggatc gtttcgcatg
      481 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
      541 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
      601 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
      661 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
      721 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
      781 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
      841 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
      901 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
      961 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     1021 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     1081 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     1141 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     1201 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     1261 gagttcttct gagcgggact ctggggttcg agagctcgct tggactcctg ttgatagatc
     1321 cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt
     1381 tttttattgg tgagaatcca agcactagtc atgaccaaaa tcccttaacg tgagttttcg
     1441 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     1501 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1561 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     1621 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1681 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1741 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1801 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     1861 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     1921 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     1981 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     2041 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     2101 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     2161 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgccc
     2221 taggtctagg gcggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata
     2281 aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct ctagcacgcg
     2341 taccacaatt ggaa
//
