LOCUS       sequence-173801-        4443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  guideRNA targeting exon2 of mouse TZAP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4443)
  AUTHORS   Li JS, Miralles Fuste J, Simavorian T, Bartocci C, Tsai J, Karlseder
            J, Lazzerini Denchi E
  TITLE     TZAP: A telomere-associated protein involved in telomere length 
            control.
  JOURNAL   Science. 2017 Jan 12. pii: aah6752. doi: 10.1126/science.aah6752.
  PUBMED    28082411
REFERENCE   2  (bases 1 to 4443)
  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: "Science. 
            2017 Jan 12. pii: aah6752. doi: 10.1126/science.aah6752."
COMMENT     Sequence Label: pCDNA-H1-sgRNA-mTZAP
FEATURES             Location/Qualifiers
     source          1..4443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(19..38)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     265..285
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(294..722)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(404..425)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     616..635
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        complement(919..1009)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     complement(979..998)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     promoter        1063..1167
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1168..1236,1237..2028)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1386..1405)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2199..2787
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2688..2707
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2941..2958
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3075..3096
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3111..3116,3117..3134,3135..3141)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3149..3165
                     /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)."
     primer_bind     3154..3176
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3173..3189
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3173..3189
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3238..3359
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3249..3268)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3303..3322
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3533..4327)
                     /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(3644..3663)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4254..4273
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(4394..4443,1..280))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(4408..4443,1..100)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gctcagaggc agaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg
       61 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga
      121 ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
      181 gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
      241 ctggggagcc tggggacttt ccacacccta actgacacac attccacaga attaattcgc
      301 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
      361 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
      421 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
      481 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
      541 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
      601 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
      661 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
      721 gtggggatac cccctagagc cccagctggt tctttccgcc tcagaagcca tagagcccac
      781 cgcatcccca gcatgcctaa aaaagcaccg actcggtgcc actttttcaa gttgataacg
      841 gactagcctt atttaaactt gctatgctgt ttccagcata gctcttaaac ttccggtcga
      901 atgtcccaca ggatctgagt ggtctcatac agaacttata agattcccaa atccaaagac
      961 atttcacgtt tatggtgatt tcccagaaca catagcgaca tgcaaatatt gcagctagct
     1021 cccgatccgt cgacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     1081 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1141 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     1201 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     1261 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     1321 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     1381 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     1441 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     1501 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     1561 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     1621 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     1681 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     1741 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     1801 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     1861 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     1921 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     1981 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     2041 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     2101 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     2161 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2221 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2281 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2341 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2401 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2461 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2521 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2581 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2641 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2701 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2761 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2821 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2881 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     2941 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     3001 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     3061 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     3121 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     3181 agctatgacc atgattacgc caagctctag ctagaggtcg acggtataca gacatgataa
     3241 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     3301 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagttg
     3361 gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc
     3421 cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt
     3481 gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc gctcagaaga
     3541 actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa
     3601 gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca
     3661 acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa
     3721 agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat
     3781 cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct
     3841 gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc
     3901 gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca
     3961 gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca
     4021 ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa
     4081 cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct
     4141 cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc
     4201 cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt
     4261 catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt
     4321 caatcatgcg aaacgatcct catcctgtct cttgatcaga tccgaaaatg gatatacaag
     4381 ctcccgggag ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga
     4441 ata
//
