LOCUS       sequence-128454-        3538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homologues recombination donor plasmid fo the wild type TERT 
            promotor region. Includes 1kb of TERT coding region in right 
            homologues arm..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3538)
  AUTHORS   Xi L, Schmidt JC, Zaug AJ, Ascarrunz DR, Cech TR
  TITLE     A novel two-step genome editing strategy with CRISPR-Cas9 provides 
            new insights into telomerase action and TERT gene expression.
  JOURNAL   Genome Biol. 2015 Nov 10;16(1):231. doi: 10.1186/s13059-015-0791-1.
  PUBMED    26553065
REFERENCE   2  (bases 1 to 3538)
  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: "Genome 
            Biol."; date: "2015-11-10"; volume: "16(1):231. doi"; pages: " 
            10.1186/s13059-015-0791-1"
COMMENT     Sequence Label: HR-TERT-promotor
FEATURES             Location/Qualifiers
     source          1..3538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1362..1378)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1362..1378)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1375..1397)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1386..1402
                     /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(1410..1416,1417..1434,1435..1440))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1455..1476
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1593..1610)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1764..2352)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1844..1863)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2523..3338)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2523..3338)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3246..3265
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3339..3443)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3511..3529
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt ccccttcacg tccggcattc
      421 gtggtgcccg gagcccgacg ccccgcgtcc ggacctggag gcagccctgg gtctccggat
      481 caggccagcg gccaaagggt cgccgcacgc acctgttccc agggcctcca catcatggcc
      541 cctccctcgg gttaccccac agcctaggcc gattcgacct ctctccgctg gggccctcgc
      601 tggcgtccct gcaccctggg agcgcgagcg gcgcgcgggc ggggaagcgc ggcccagacc
      661 cccgggtccg cccggagcag ctgcgctgtc ggggccaggc cgggctccca gtggattcgc
      721 gggcacagac gcccaggacc gcgcttccca cgtggcggag ggactgggga cccgggcacc
      781 cgtcctgccc cttcaccttc cagctccgcc tcctccgcgc ggaccccgcc ccgtcccgac
      841 ccctcccggg tccccggccc agccccctcc gggccctccc agcccctccc cttcctttcc
      901 gcggccccgc cctctcctcg cggcgcgagt ttcaggcagc gctgcgtcct gctgcgcacg
      961 tgggaagccc tggccccggc cacccccgcg atgccgcgcg ctccccgctg ccgagccgtg
     1021 cgctccctgc tgcgcagcca ctaccgcgag gtgctgccgc tggccacgtt cgtgcggcgc
     1081 ctggggcccc agggctggcg gctggtgcag cgcggggacc cggcggcttt ccgcgcgctg
     1141 gtggcccagt gcctggtgtg cgtgccctgg gacgcacggc cgccccccgc cgccccctcc
     1201 ttccgccagg tgggcctccc cggggtcggc gtccggctgg ggttgagggc ggccgggggg
     1261 aaccagcgac atgcggagag cagcgcaggc gactcagggc gcttcccccg caggtgtcct
     1321 gcctgaagga gctggtggcc cgaaagcttg gctcgagcat ggtcatagct gtttcctgtg
     1381 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     1441 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     1501 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     1561 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     1621 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1681 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1741 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1801 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1861 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1921 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1981 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2041 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2101 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2161 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2221 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2281 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2341 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2401 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2461 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2521 agttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     2581 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     2641 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     2701 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     2761 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     2821 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     2881 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     2941 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     3001 tattcttcta atacctggaa tgctgttttc ccagggatcg cagtggtgag taaccatgca
     3061 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     3121 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     3181 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     3241 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     3301 ggcctagagc aagacgtttc ccgttgaata tggctcatac tcttcctttt tcaatattat
     3361 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     3421 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     3481 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
