LOCUS       sequence-380412-        5867 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor template for insertion of hTERT promoter (-146 C to T) driven 
            Gaussia luciferase reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5867)
  AUTHORS   Sharma S, Mukherjee AK, Roy SS, Bagri S, Lier S, Verma M, Sengupta 
            A, Kumar M, Nesse G, Pandey DP, Chowdhury S
  TITLE     Human telomerase is directly regulated by non-telomeric 
            TRF2-G-quadruplex interaction.
  JOURNAL   Cell Rep. 2021 May 18;35(7):109154. doi: 
            10.1016/j.celrep.2021.109154.
  PUBMED    34010660
REFERENCE   2  (bases 1 to 5867)
  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: "2021-05-18"; volume: "35(7):109154. doi"; pages: " 
            10.1016/j.celrep.2021.109154"
COMMENT     Sequence Label: TERTp146_Gluc_CCR5
FEATURES             Location/Qualifiers
     source          1..5867
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(38..60)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     160..179
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     373..395
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     387..404
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     388..404
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    558..569
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     regulatory      2293..2302
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2299..2856
                     /codon_start=1
                     /product="enhanced variant of secreted Gaussia luciferase"
                     /label=Gaussia-Dura Luc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPLEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     CDS             2299..2856
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPLEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     polyA_signal    2880..3001
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2891..2910)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2945..2964
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3655..3671)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3655..3671)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3668..3690)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3679..3695
                     /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(3703..3709,3710..3727,3728..3733))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3748..3769
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3886..3903)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4057..4645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4137..4156)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4816..5607,5608..5676))
                     /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     5439..5458
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5677..5781)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5849..5867
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
       61 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
      121 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
      181 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
      241 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
      301 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
      361 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattc gagctcggta
      421 cctcgcgaat gcatctagat gacgaccgcc ggcgtttgct gctagcttcc ctgtccacta
      481 gatggagagc ttggctctgt tgggggaatt catgaaagca ccatctgttt aaacttcagc
      541 cttttgcagt ttatcaggat gaggatgacc agcatgttgc ccacaaaacc aaagatgaac
      601 accagtgagt agagcggagg caggaggcgg gctgcgattt gcttcacatt gattttttgg
      661 cagggctccg atgtataata attgatgtca tagattggac ttgacacttg ataatccatc
      721 ttgttccacc ctgtgcataa ataaaaagtg atcttttata aagtcctaga atgtatttag
      781 ttgccctcca tgaatgcaaa ctgttttata catcaatagg tttttaattg cctacataga
      841 tgtctacatt gaattaactc tctttttggc caagcaatga agttttgtag tgaagggaag
      901 gtttgctgct agcttccctg tccactagat ggagaagctt gttaacacgc gtagatctca
      961 gcccagggag ggtgcgaggc ctgttcaaat gctagctcca taaataaagc aatttcctcc
     1021 ggcagtttct gaaagtagga aaggttacat ttaaggttgc gtttgttagc atttcagtgt
     1081 ttgccgacct cagctacagc atccctgcaa ggcctcggga gacccagaag tttctcgccc
     1141 cttagatcca aacttgagca acccggagtc tggattcctg ggaagtcctc agctgtcctg
     1201 cggttgtgcc ggggccccag gtctggaggg gaccagtggc cgtgtggctt ctactgctgg
     1261 gctggaagtc gggcctccta gctctgcagt ccgaggcttg gagccaggtg cctggacccc
     1321 gaggctgccc tccaccctgt gcgggcggga tgtgaccaga tgttggcctc atctgccaga
     1381 cagagtgccg gggcccaggg tcaaggccgt tgtggctggt gtgaggcgcc cggtgcgcgg
     1441 ccagcaggag cgcctggctc catttcccac cctttctcga cgggaccgcc ccggtgggtg
     1501 attaacagat ttggggtggt ttgctcatgg tggggacccc tcgccgcctg agaacctgca
     1561 aagagaaatg acgggcctgt gtcaaggagc ccaagtcgcg gggaagtgtt gcagggaggc
     1621 actccgggag gtcccgcgtg cccgtccagg gagcaatgcg tcctcgggtt cgtccccagc
     1681 cgcgtctacg cgcctccgtc ctccccttca cgtccggcat tcgtggtgcc cggagcccga
     1741 cgccccgcgt ccggacctgg aggcagccct gggtctccgg atcaggccag cggccaaagg
     1801 gtcgccgcac gcacctgttc ccagggcctc cacatcatgg cccctccctc gggttacccc
     1861 acagcctagg ccgattcgac ctctctccgc tggggccctc gctggcgtcc ctgcaccctg
     1921 ggagcgcgag cggcgcgcgg gcggggaagc gcggcccaga cccccgggtc cgcccggagc
     1981 agctgcgctg tcggggccag gccgggctcc cagtggattc gcgggcacag acgcccagga
     2041 ccgcgcttcc cacgtggcgg agggactggg gacccgggca cccgtcctgc cccttcacct
     2101 tccagctccg cctcctccgc gcggaccccg ccccgtcccg accccttccg ggtccccggc
     2161 ccagccccct ccgggccctc ccagcccctc cccttccttt ccgcggcccc gccctctcct
     2221 cgcggcgcga gtttcaggca gcgctgcgtc ctgctgcgca cgtgggaaag cttggtaccg
     2281 agctcggatc cagccaccat gggagtcaaa gttctgtttg ccctgatctg catcgctgtg
     2341 gccgaggcca agcccaccga gaacaacgaa gacttcaaca tcgtggccgt ggccagcaac
     2401 ttcgcgacca cggatctcga tgctgaccgc gggaagttgc ccggcaagaa gctgccgctg
     2461 gaggtgctca aagagctgga agccaatgcc cggaaagctg gctgcaccag gggctgtctg
     2521 atctgcctgt cccacatcaa gtgcacgccc aagatgaaga agttcatccc aggacgctgc
     2581 cacacctacg aaggcgacaa agagtccgca cagggcggca taggcgaggc gatcgtcgac
     2641 attcctgaga ttcctgggtt caaggacttg gagcccctgg agcagttcat cgcacaggtc
     2701 gatctgtgtg tggactgcac aactggctgc ctcaaagggc ttgccaacgt gcagtgttct
     2761 gacctgctca agaagtggct gccgcaacgc tgtgcgacct ttgccagcaa gatccagggc
     2821 caggtggaca agatcaaggg ggccggtggt gactaagcgg ccgcttcgag cagacatgat
     2881 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     2941 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     3001 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     3061 ttaaagcaag taaaacctct acaaatgtgg tactcgagac gcgtgctctg ttgggggaat
     3121 tcatgaaagc accatctcac caaataaaat cttgtgctct atagcaccat ggagtgaatg
     3181 aagctttgac aacaatttta agaaactggg tcaagcatgg tgcagctcag cctttttgtt
     3241 gttgttgttg ttgttgtgag acagggtctc actctgtcgc ccaggctgga gtgcagtggc
     3301 acgatcatgg ctcactgcag cctggaccat cccgggctca agcaatcctc tcacctcacc
     3361 cctccaagtg gctggggact acaggcgcca ccacaccaag ctaattttgt ttatttttat
     3421 agatacaaga tcacactatg ttgcccaggc tggtctcgaa ctcctgggct caagcgatcc
     3481 tcctgcctca gcctcctaga atgcatttaa attttgctgc tagcttccct gtccactaga
     3541 tggagagctt ggctctgttg ggggaattca tgaaagcacc atctgcggcc gcgtcgtatc
     3601 ggatcccggg cccgtcgact gcagaggcct gcatgcaagc ttggcgtaat catggtcata
     3661 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3721 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3781 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3841 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3901 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3961 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4021 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4081 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4141 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4201 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4261 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4321 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4381 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4441 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4501 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4561 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4621 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4681 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4741 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4801 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4861 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4921 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4981 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5041 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5101 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5161 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5221 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5281 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5341 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5401 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5461 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5521 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5581 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5641 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5701 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5761 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     5821 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggc
//
