LOCUS       sequence-128453-        4596 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homologues recombination donor plasmid fo the TERT promotor region 
            introducing a SV-40 driven GFP marker in the TERT promotor. 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 4596)
  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 4596)
  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-SV40-GFP
FEATURES             Location/Qualifiers
     source          1..4596
                     /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(838..858)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        843..1172
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1023..1158
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(1179..1181,1182..1184,1185..1898)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1179..1181,1182..1184,1185..1895)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1224..1245)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1485..1504)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1832..1853
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1899..1947
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2420..2436)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2420..2436)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2433..2455)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2444..2460
                     /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(2468..2474,2475..2492,2493..2498))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2513..2534
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2651..2668)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2822..3410)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2902..2921)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3581..4396)
                     /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(3581..4396)
                     /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     4304..4323
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4397..4501)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4569..4587
                     /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 cccggagccc gacgccccgc
      421 gtccggacct ggaggcagcc ctgggtctcc ggatcaggcc agcggccaaa gggtcgccgc
      481 acgcacctgt tcccagggcc tccacatcat ggcccctccc tcgggttacc ccacagccta
      541 ggccgattcg acctctctcc gctggggccc tcgctggcgt ccctgcaccc tgggagcgcg
      601 agcggcgcgc gggcggggaa gcgcggccca gacccccggg tccgcccgga gcagctgcgc
      661 tgtcggggcc aggccgggct cccagtggat tcgcgggcac agacgcccag gaccgcgctt
      721 cccacgtggc ggagggactg gggacccggg cacccgtcct gccccttcac cttccagctc
      781 cgcctcctcc gcgcggaccc cgccccgtcc cgacccctcc cgggtggtcg accctgtgga
      841 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
      901 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
      961 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     1021 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     1081 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     1141 gaggcttttt tggaggccta ggcttttgca aaaagctcat ggtgagcaag ggcgaggagc
     1201 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1261 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     1321 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     1381 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1441 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     1501 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1561 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1621 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     1681 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1741 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     1801 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1861 ccgggatcac tctcggcatg gacgagctgt acaagtaaaa taaaatatct ttattttcat
     1921 tacatctgtg tgttggtttt ttgtgtgagg atctaaatga gtcttcggac ctcgcggccc
     1981 cggcccagcc ccctccgggc cctcccagcc cctccccttc ctttccgcgg ccccgccctc
     2041 tcctcgcggc gcgagtttca ggcagcgctg cgtcctgctg cgcacgtggg aagccctggc
     2101 cccggccacc cccgcgatgc cgcgcgctcc ccgctgccga gccgtgcgct ccctgctgcg
     2161 cagccactac cgcgaggtgc tgccgctggc cacgttcgtg cggcgcctgg ggccccaggg
     2221 ctggcggctg gtgcagcgcg gggacccggc ggctttccgc gcgctggtgg cccagtgcct
     2281 ggtgtgcgtg ccctgggacg cacggccgcc ccccgccgcc ccctccttcc gccaggtggg
     2341 cctccccggg gtcggcgtcc ggctggggtt gagggcggcc ggggggaacc agcgacatgc
     2401 gaagcttggc tcgagcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     2461 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     2521 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2581 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2641 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2701 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2761 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2821 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2881 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2941 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3001 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3061 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3121 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3181 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3241 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3301 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3361 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3421 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3481 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3541 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttagaaaaac tcatcgagca
     3601 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     3661 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     3721 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     3781 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     3841 aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     3901 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
     3961 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     4021 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     4081 ctgttttccc agggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     4141 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     4201 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     4261 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     4321 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa gacgtttccc
     4381 gttgaatatg gctcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4441 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4501 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     4561 cctataaaaa taggcgtatc acgaggccct ttcgtc
//
