LOCUS       sequence-173874-        4443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  guideRNA targeting exon1 of human 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-hTZAP
FEATURES             Location/Qualifiers
     source          1..4443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    287..408
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(324..343)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     378..397
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(457..473)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(457..473)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(470..492)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    481..497
                     /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(505..511,512..529,530..535))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    550..571
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(688..705)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(859..1447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(939..958)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1618..2409,2410..2478))
                     /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     2241..2260
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2479..2583)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2637..2727
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     2648..2667
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     rep_origin      2924..3352
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3011..3030)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3221..3242
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3361..3381)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3366..3695
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3546..3681
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3608..3627
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             join(3762..4443,1..113)
                     /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(3816..3835)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     join(4426..4443,1..2)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
       61 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
      121 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
      181 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
      241 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
      301 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
      361 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
      421 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
      481 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
      541 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
      601 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
      661 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
      721 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      781 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      841 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      901 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      961 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1021 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1081 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1141 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1201 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1261 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1321 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1381 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1441 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1501 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1561 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1621 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1681 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1741 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     1801 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1861 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1921 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1981 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     2041 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     2101 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2161 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2221 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2281 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2341 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2401 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2461 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2521 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2581 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagcta gctgcaatat
     2641 ttgcatgtcg ctatgtgttc tgggaaatca ccataaacgt gaaatgtctt tggatttggg
     2701 aatcttataa gttctgtatg agaccactca gatcccttcg tccagcacag tgtgagttta
     2761 agagctatgc tggaaacagc atagcaagtt taaataaggc tagtccgtta tcaacttgaa
     2821 aaagtggcac cgagtcggtg cttttttagg catgctgggg atgcggtggg ctctatggct
     2881 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     2941 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3001 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3061 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3121 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3181 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3241 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3301 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3361 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3421 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3481 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3541 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3601 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3661 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     3721 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3781 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3841 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3901 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     3961 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4021 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4081 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4141 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4201 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4261 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4321 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4381 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4441 ata
//
