LOCUS       sequence-370669-        3565 bp    DNA     circular SYN 15-APR-2025
DEFINITION  sgRNA plasmid encoding 7SK promoter driving an anti-safe targeting 
            sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3565)
  AUTHORS   Brewer WJ, Xet-Mull AM, Yu A, Sweeney MI, Walton EM, Tobin DM
  TITLE     Macrophage NFATC2 mediates angiogenic signaling during mycobacterial
            infection.
  JOURNAL   Cell Rep. 2022 Dec 13;41(11):111817. doi: 
            10.1016/j.celrep.2022.111817.
  PUBMED    36516756
REFERENCE   2  (bases 1 to 3565)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep.";
            date: "2022-12-13"; volume: "41(11):111817. doi"; pages: " 
            10.1016/j.celrep.2022.111817"
COMMENT     Sequence Label: p7SK ST
FEATURES             Location/Qualifiers
     source          1..3565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        328..403
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(459..479)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(459..477)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(498..514)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(498..514)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(511..533)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    522..538
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(546..552,553..570,571..576))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    591..612
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(729..746)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(900..1488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(980..999)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1625..2419)
                     /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"
     CDS             complement(1625..2419)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1736..1755)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2346..2365
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2454..2811)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2468..2603
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        2620..2811
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     2768..2788
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(2813..2917)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2943..3398)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3080..3101)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3292..3311
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3525..3547
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3539..3556
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3540..3556
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattgggt acctttaatt ctagtactat gcatcgtctc
       61 attgtctgca gtatttagca tgccccaccc atctgcaagg cattctggat agtgtcaaaa
      121 cagccggaaa tcaagtccgt ttatctcaaa ctttagcatt ttgggaataa atgatatttg
      181 ctatgctggt taaattagat tttagttaaa tttcctgctg aagctctagt acgataagca
      241 acttgaccta agtgtaaagt tgagacttcc ttcaggttta tatagcttgt gcgccgcttg
      301 ggtacctcgg tggataactt cctgagtgtt ttagagctag aaatagcaag ttaaaataag
      361 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt ctcggactga
      421 tgacctgaga cgccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg
      481 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
      541 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
      601 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
      661 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
      721 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
      781 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
      841 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
      901 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
      961 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1021 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     1081 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     1141 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     1201 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     1261 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     1321 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     1381 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     1441 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     1501 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     1561 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcatc
     1621 atgatcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg
     1681 cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat
     1741 cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga
     1801 tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg
     1861 tcacgacgag atcctcgccg tcgggcatgc tcgccttgag cctggcgaac agttcggctg
     1921 gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc
     1981 gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat
     2041 caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa
     2101 ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg
     2161 cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata
     2221 gccgcgctgc ctcgtcttgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa
     2281 gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct
     2341 gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca
     2401 atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatcg atctttgcaa
     2461 aagcctaggc cttcaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg
     2521 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
     2581 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
     2641 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
     2701 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
     2761 tttccacacc ctaactgaca cacattccac agctggttct ttccgcctca ggactcttcc
     2821 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     2881 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     2941 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3001 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3061 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3121 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3181 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3241 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3301 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3361 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
     3421 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     3481 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     3541 taaaacgacg gccagtgagc gcgcg
//
