LOCUS       sequence-379160-        2354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded plasmid with BsaI sites to clone sgRNA. The 6-nucleotide 
            barcode (sequence: GCATGG) 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_BC3
FEATURES             Location/Qualifiers
     source          1..2354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(286..304)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             483..1277
                     /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(537..556)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1147..1166
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      1308..1402
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1490..2078
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1979..1998
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2243..2329)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tggaacccga cgaacgtgta tgtttcacta cgtgtggcgt cctggacagg ggatcgctta
       61 acaccaactg gttttcaggc gatgggtagc cgcagcacag tgaagacacg gctatccata
      121 ctagatagag aacctccgat aatgctgcta ccgttttcta atcttgacga tctgtcggcc
      181 attattagta cagcgaggca acccatgcac ggatccccac cgagaccttt ggtctcaacc
      241 tctcgaggtg aagacgaaag gggcatacat aacatggagt cgtacgcctc gtgatacgcc
      301 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcggaa ttgccagctg
      361 gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc
      421 caaggatctg atggcgcagg ggatcaagat ctgatcaaga gacaggatga ggatcgtttc
      481 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
      541 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
      601 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
      661 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
      721 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
      781 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
      841 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
      901 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
      961 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     1021 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     1081 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     1141 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     1201 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     1261 ttgacgagtt cttctgagcg ggactctggg gttcgagagc tcgcttggac tcctgttgat
     1321 agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg
     1381 ggcgtttttt attggtgaga atccaagcac tagtcatgac caaaatccct taacgtgagt
     1441 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1501 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1561 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     1621 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     1681 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     1741 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     1801 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     1861 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     1921 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     1981 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2041 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2101 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     2161 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     2221 cgccctaggt ctagggcggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
     2281 agataaaacg aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctctagc
     2341 acgcgtacca caat
//
