LOCUS       sequence-379164-        2354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded plasmid with BsaI sites to clone sgRNA. The 6-nucleotide 
            barcode (sequence: AGTCTA) 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_BC5
FEATURES             Location/Qualifiers
     source          1..2354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(274..292)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             471..1265
                     /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(525..544)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1135..1154
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      1296..1390
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1478..2066
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1967..1986
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2231..2317)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 aacgtgtatg tttcactacg tgtggcgtcc tggacagggg atcgcttaac accaactggt
       61 tttcaggcga tgggtagccg cagcacagtg aagacacggc tatccatact agatagagaa
      121 cctccgataa tgctgctacc gttttctaat cttgacgatc tgtcggccat tattagtaca
      181 gcgaggcaac tagactacgg atccccaccg agacctttgg tctcaacctc tcgaggtgaa
      241 gacgaaaggg gcatacataa catggagtcg tacgcctcgt gatacgccta tttttatagg
      301 ttaatgtcat gataataatg gtttcttaga cgtcggaatt gccagctggg gcgccctctg
      361 gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat
      421 ggcgcagggg atcaagatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
      481 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
      541 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
      601 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
      661 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
      721 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
      781 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
      841 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
      901 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
      961 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     1021 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     1081 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     1141 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     1201 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     1261 tctgagcggg actctggggt tcgagagctc gcttggactc ctgttgatag atccagtaat
     1321 gacctcagaa ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat
     1381 tggtgagaat ccaagcacta gtcatgacca aaatccctta acgtgagttt tcgttccact
     1441 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1501 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1561 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1621 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1681 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1741 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     1801 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     1861 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     1921 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     1981 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2041 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2101 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2161 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg ccctaggtct
     2221 agggcggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa
     2281 aggcccagtc tttcgactga gcctttcgtt ttatttgatg cctctagcac gcgtaccaca
     2341 attggaaccc gacg
//
