LOCUS       sequence-201861-        2700 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Can be used as PCR template for a STAgR reaction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2700)
  AUTHORS   Breunig CT, Durovic T, Neuner AM, Baumann V, Wiesbeck MF, Koferle A,
            Gotz M, Ninkovic J, Stricker SH
  TITLE     One step generation of customizable gRNA vectors for multiplex 
            CRISPR approaches through string assembly gRNA cloning (STAgR).
  JOURNAL   PLoS One. 2018 Apr 27;13(4):e0196015. doi: 
            10.1371/journal.pone.0196015. eCollection 2018.
  PUBMED    29702666
REFERENCE   2  (bases 1 to 2700)
  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"; journalName: "PLoS One.";
            date: "2018-04-27"; volume: "13(4):e0196015. doi"; pages: " 
            10.1371/journal.pone.0196015. eCollection 2018"
COMMENT     Sequence Label: STAGR_SAMScaffold_hU6
FEATURES             Location/Qualifiers
     source          1..2700
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(474..1268)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(1269..1373)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1674..1696
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1688..1705
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1689..1705
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1716..1735
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1716..1734
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1739..1750
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_RNA        1783..1801
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        1853..1871
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     promoter        1938..2186
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1938..1958
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2109..2128
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(2302..2319)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(2412..2700,1..300))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2492..2511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
       61 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
      121 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
      181 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
      241 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
      301 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
      361 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      421 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttattagaaa
      481 aattcatcca gcagacgata aaacgcaata cgctggctat ccggtgccgc aatgccatac
      541 agcaccagaa aacgatccgc ccattcgccg cccagttctt ccgcaatatc acgggtggcc
      601 agcgcaatat cctgataacg atccgccacg cccagacggc cgcaatcaat aaagccgcta
      661 aaacggccat tttccaccat aatgttcggc aggcacgcat caccatgggt caccaccaga
      721 tcttcgccat ccggcatgct cgctttcaga cgcgcaaaca gctctgccgg tgccaggccc
      781 tgatgttctt catccagatc atcctgatcc accaggcccg cttccatacg ggtacgcgca
      841 cgttcaatac gatgtttcgc ctgatgatca aacggacagg tcgccgggtc cagggtatgc
      901 agacgacgca tggcatccgc cataatgctc actttttctg ccggcgccag atggctagac
      961 agcagatcct gacccggcac ttcgcccagc agcagccaat cacggcccgc ttcggtcacc
     1021 acatccagca ccgccgcaca cggaacaccg gtggtggcca gccagctcag acgcgccgct
     1081 tcatcctgca gctcgttcag cgcaccgctc agatcggttt tcacaaacag caccggacga
     1141 ccctgcgcgc tcagacgaaa caccgccgca tcagagcagc caatggtctg ctgcgcccaa
     1201 tcatagccaa acagacgttc cacccacgct gccgggctac ccgcatgcag gccatcctgt
     1261 tcaatcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     1321 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     1381 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     1441 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     1501 caaaagaata gaccgagata gggttgagtg gccgctacag ggcgctccca ttcgccattc
     1561 aggctgcgca actgttggga agggcgtttc ggtgcgggcc tcttcgctat tacgccagct
     1621 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     1681 acgacgttgt aaaacgacgg ccagtgagcg cgacgtaata cgactcacta tagggcgaat
     1741 tgaaggaagg ccgtcaaggc cgcatgtttt agagctaggc caacatgagg atcacccatg
     1801 tctgcagggc ctagcaagtt aaaataaggc tagtccgtta tcaacttggc caacatgagg
     1861 atcacccatg tctgcagggc caagtggcac cgagtcggtg ctttttttcg gtgctttttt
     1921 ttaaggtcgg gcaggaagag ggcctatttc ccatgattcc ttcatatttg catatacgat
     1981 acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag atattagtac
     2041 aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt
     2101 ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat
     2161 atatcttgtg gaaaggacga aacaccgctg ggcctcatgg gccttccttt cactgcccgc
     2221 tttccagtcg ggaaacctgt cgtgccagct gcattaacat ggtcatagct gtttccttgc
     2281 gtattgggcg ctctccgctt cctcgctcac tgactcgctg cgctcggtcg ttcgggtaaa
     2341 gcctggggtg cctaatgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2401 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2461 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2521 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2581 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2641 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
//
