LOCUS       sequence-381604-        2354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded plasmid with BsaI sites to clone sgRNA. The 6-nucleotide 
            barcode (sequence: ATCATG) 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_BC2
FEATURES             Location/Qualifiers
     source          1..2354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(279..297)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             476..1270
                     /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(530..549)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1140..1159
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      1301..1395
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1483..2071
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1972..1991
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2236..2322)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cgacgaacgt gtatgtttca ctacgtgtgg cgtcctggac aggggatcgc ttaacaccaa
       61 ctggttttca ggcgatgggt agccgcagca cagtgaagac acggctatcc atactagata
      121 gagaacctcc gataatgctg ctaccgtttt ctaatcttga cgatctgtcg gccattatta
      181 gtacagcgag gcaaccatga tacggatccc caccgagacc tttggtctca acctctcgag
      241 gtgaagacga aaggggcata cataacatgg agtcgtacgc ctcgtgatac gcctattttt
      301 ataggttaat gtcatgataa taatggtttc ttagacgtcg gaattgccag ctggggcgcc
      361 ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat
      421 ctgatggcgc aggggatcaa gatctgatca agagacagga tgaggatcgt ttcgcatgat
      481 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
      541 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
      601 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
      661 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
      721 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
      781 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
      841 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
      901 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
      961 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     1021 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     1081 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     1141 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     1201 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     1261 gttcttctga gcgggactct ggggttcgag agctcgcttg gactcctgtt gatagatcca
     1321 gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt
     1381 tttattggtg agaatccaag cactagtcat gaccaaaatc ccttaacgtg agttttcgtt
     1441 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1501 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1561 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     1621 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     1681 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     1741 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     1801 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     1861 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     1921 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     1981 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2041 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2101 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     2161 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgcccta
     2221 ggtctagggc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
     2281 acgaaaggcc cagtctttcg actgagcctt tcgttttatt tgatgcctct agcacgcgta
     2341 ccacaattgg aacc
//
