LOCUS       sequence-370674-        3452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA plasmid encoding hH1 promoter driving an anti-safe targeting 
            sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3452)
  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 3452)
  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: "Cell Rep.";
            date: "2022-12-13"; volume: "41(11):111817. doi"; pages: " 
            10.1016/j.celrep.2022.111817"
COMMENT     Sequence Label: phH1 ST
FEATURES             Location/Qualifiers
     source          1..3452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..215
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     136..155
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     misc_RNA        243..318
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(363..383)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(363..381)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(402..418)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(402..418)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(415..437)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    426..442
                     /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(450..456,457..474,475..480))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    495..516
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(633..650)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(804..1392)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(884..903)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1469..2263)
                     /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(1469..2263)
                     /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(1580..1599)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2190..2209
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2298..2655)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2312..2447
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        2464..2655
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     2612..2632
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(2657..2761)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2787..3242)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2924..2945)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3136..3155
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3369..3391
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3383..3400
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3384..3400
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3410..3429
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3410..3428
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gaacgctgac gtcatcaacc cgctccaagg aatcgcgggc ccagtgtcac taggcgggaa
       61 cacccagcgc gcgtgcgccc tggcaggaag atggctgtga gggacagggg agtggcgccc
      121 tgcaatattt gcatgtcgct atgtgttctg ggaaatcacc ataaacgtga aatgtctttg
      181 gatttgggaa tcttataagt tctgtatgag accactcttt cccagtgcag ttctccgggt
      241 tggttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa
      301 gtggcaccga gtcggtgctt tttttggtct gagacgccgc ggtggagctc cagcttttgt
      361 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
      421 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
      481 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
      541 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
      601 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
      661 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
      721 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      781 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      841 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      901 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
      961 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1021 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1081 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1141 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1201 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1261 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1321 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1381 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1441 aactcacgtt aagggatttt ggtcatgatc agaagaactc gtcaagaagg cgatagaagg
     1501 cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt
     1561 cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg
     1621 ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat
     1681 tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgctcgcct
     1741 tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct
     1801 gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt
     1861 ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga
     1921 tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc
     1981 ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa
     2041 cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac
     2101 cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg
     2161 cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc
     2221 aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc
     2281 ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
     2341 tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
     2401 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
     2461 ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     2521 gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt
     2581 ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgg
     2641 ttctttccgc ctcaggactc ttcctttttc aatattattg aagcatttat cagggttatt
     2701 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     2761 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     2821 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     2881 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     2941 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     3001 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     3061 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     3121 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     3181 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     3241 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     3301 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     3361 agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc
     3421 actatagggc gaattgggta cccgtctcac ct
//
