LOCUS       sequence-380414-        5867 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor template for insertion of hTERT promoter 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: TERTpWT_Gluc_CCR5
FEATURES             Location/Qualifiers
     source          1..5867
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(35..57)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     157..176
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     370..392
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     384..401
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     385..401
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    555..566
                     /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      2290..2299
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2296..2853
                     /codon_start=1
                     /product="enhanced variant of secreted Gaussia luciferase"
                     /label=Gaussia-Dura Luc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPLEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     CDS             2296..2853
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPLEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     polyA_signal    2877..2998
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2888..2907)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2942..2961
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3652..3668)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3652..3668)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3665..3687)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3676..3692
                     /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(3700..3706,3707..3724,3725..3730))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3745..3766
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3883..3900)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4054..4642)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4134..4153)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4813..5604,5605..5673))
                     /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     5436..5455
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5674..5778)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5846..5864
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
       61 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
      121 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
      181 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
      241 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
      301 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
      361 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattcgag ctcggtacct
      421 cgcgaatgca tctagatgac gaccgccggc gtttgctgct agcttccctg tccactagat
      481 ggagagcttg gctctgttgg gggaattcat gaaagcacca tctgtttaaa cttcagcctt
      541 ttgcagttta tcaggatgag gatgaccagc atgttgccca caaaaccaaa gatgaacacc
      601 agtgagtaga gcggaggcag gaggcgggct gcgatttgct tcacattgat tttttggcag
      661 ggctccgatg tataataatt gatgtcatag attggacttg acacttgata atccatcttg
      721 ttccaccctg tgcataaata aaaagtgatc ttttataaag tcctagaatg tatttagttg
      781 ccctccatga atgcaaactg ttttatacat caataggttt ttaattgcct acatagatgt
      841 ctacattgaa ttaactctct ttttggccaa gcaatgaagt tttgtagtga agggaaggtt
      901 tgctgctagc ttccctgtcc actagatgga gaagcttgtt aacacgcgta gatctcagcc
      961 cagggagggt gcgaggcctg ttcaaatgct agctccataa ataaagcaat ttcctccggc
     1021 agtttctgaa agtaggaaag gttacattta aggttgcgtt tgttagcatt tcagtgtttg
     1081 ccgacctcag ctacagcatc cctgcaaggc ctcgggagac ccagaagttt ctcgcccctt
     1141 agatccaaac ttgagcaacc cggagtctgg attcctggga agtcctcagc tgtcctgcgg
     1201 ttgtgccggg gccccaggtc tggaggggac cagtggccgt gtggcttcta ctgctgggct
     1261 ggaagtcggg cctcctagct ctgcagtccg aggcttggag ccaggtgcct ggaccccgag
     1321 gctgccctcc accctgtgcg ggcgggatgt gaccagatgt tggcctcatc tgccagacag
     1381 agtgccgggg cccagggtca aggccgttgt ggctggtgtg aggcgcccgg tgcgcggcca
     1441 gcaggagcgc ctggctccat ttcccaccct ttctcgacgg gaccgccccg gtgggtgatt
     1501 aacagatttg gggtggtttg ctcatggtgg ggacccctcg ccgcctgaga acctgcaaag
     1561 agaaatgacg ggcctgtgtc aaggagccca agtcgcgggg aagtgttgca gggaggcact
     1621 ccgggaggtc ccgcgtgccc gtccagggag caatgcgtcc tcgggttcgt ccccagccgc
     1681 gtctacgcgc ctccgtcctc cccttcacgt ccggcattcg tggtgcccgg agcccgacgc
     1741 cccgcgtccg gacctggagg cagccctggg tctccggatc aggccagcgg ccaaagggtc
     1801 gccgcacgca cctgttccca gggcctccac atcatggccc ctccctcggg ttaccccaca
     1861 gcctaggccg attcgacctc tctccgctgg ggccctcgct ggcgtccctg caccctggga
     1921 gcgcgagcgg cgcgcgggcg gggaagcgcg gcccagaccc ccgggtccgc ccggagcagc
     1981 tgcgctgtcg gggccaggcc gggctcccag tggattcgcg ggcacagacg cccaggaccg
     2041 cgcttcccac gtggcggagg gactggggac ccgggcaccc gtcctgcccc ttcaccttcc
     2101 agctccgcct cctccgcgcg gaccccgccc cgtcccgacc cctcccgggt ccccggccca
     2161 gccccctccg ggccctccca gcccctcccc ttcctttccg cggccccgcc ctctcctcgc
     2221 ggcgcgagtt tcaggcagcg ctgcgtcctg ctgcgcacgt gggaaagctt ggtaccgagc
     2281 tcggatccag ccaccatggg agtcaaagtt ctgtttgccc tgatctgcat cgctgtggcc
     2341 gaggccaagc ccaccgagaa caacgaagac ttcaacatcg tggccgtggc cagcaacttc
     2401 gcgaccacgg atctcgatgc tgaccgcggg aagttgcccg gcaagaagct gccgctggag
     2461 gtgctcaaag agctggaagc caatgcccgg aaagctggct gcaccagggg ctgtctgatc
     2521 tgcctgtccc acatcaagtg cacgcccaag atgaagaagt tcatcccagg acgctgccac
     2581 acctacgaag gcgacaaaga gtccgcacag ggcggcatag gcgaggcgat cgtcgacatt
     2641 cctgagattc ctgggttcaa ggacttggag cccctggagc agttcatcgc acaggtcgat
     2701 ctgtgtgtgg actgcacaac tggctgcctc aaagggcttg ccaacgtgca gtgttctgac
     2761 ctgctcaaga agtggctgcc gcaacgctgt gcgacctttg ccagcaagat ccagggccag
     2821 gtggacaaga tcaagggggc cggtggtgac taagcggccg cttcgagcag acatgataag
     2881 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     2941 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     3001 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     3061 aagcaagtaa aacctctaca aatgtggtac tcgagacgcg tgctctgttg ggggaattca
     3121 tgaaagcacc atctcaccaa ataaaatctt gtgctctata gcaccatgga gtgaatgaag
     3181 ctttgacaac aattttaaga aactgggtca agcatggtgc agctcagcct ttttgttgtt
     3241 gttgttgttg ttgtgagaca gggtctcact ctgtcgccca ggctggagtg cagtggcacg
     3301 atcatggctc actgcagcct ggaccatccc gggctcaagc aatcctctca cctcacccct
     3361 ccaagtggct ggggactaca ggcgccacca caccaagcta attttgttta tttttataga
     3421 tacaagatca cactatgttg cccaggctgg tctcgaactc ctgggctcaa gcgatcctcc
     3481 tgcctcagcc tcctagaatg catttaaatt ttgctgctag cttccctgtc cactagatgg
     3541 agagcttggc tctgttgggg gaattcatga aagcaccatc tgcggccgcg tcgtatcgga
     3601 tcccgggccc gtcgactgca gaggcctgca tgcaagcttg gcgtaatcat ggtcatagct
     3661 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3721 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3781 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3841 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3901 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3961 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4021 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4081 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4141 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4201 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4261 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4321 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4381 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4441 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4501 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4561 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4621 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4681 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4741 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4801 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4861 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4921 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4981 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5041 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5101 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5161 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5221 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5281 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5341 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5401 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5461 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5521 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5581 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5641 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5701 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5761 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     5821 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccct
//
