LOCUS       sequence-337174-        7796 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly-Renilla luciferase assay for hTERT promoter activity of RE2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7796)
  AUTHORS   Penev A, Bazley A, Shen M, Boeke JD, Savage SA, Sfeir A
  TITLE     Alternative splicing is a developmental switch for hTERT expression.
  JOURNAL   Mol Cell. 2021 Apr 6. pii: S1097-2765(21)00228-8. doi: 
            10.1016/j.molcel.2021.03.033.
  PUBMED    33852895
REFERENCE   2  (bases 1 to 7796)
  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: "Mol Cell. 
            2021 Apr 6. pii: S1097-2765(21)00228-8. doi: 
            10.1016/j.molcel.2021.03.033."
COMMENT     Sequence Label: pRF TpC RE2
FEATURES             Location/Qualifiers
     source          1..7796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1358..1374
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1359..1375
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1389..2324)
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     promoter        complement(2372..2390)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2433..2452)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          2435..2567
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(2599..2795)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2613..2748
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     misc_feature    2850..2941
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(2882..2901)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    2955..3003
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(3134..3589)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3271..3292)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3483..3502
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3616..3720
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3721..3789,3790..4581)
                     /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     complement(3939..3958)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4752..5340
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5241..5260
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5494..5511
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5663..5683)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    5984..6105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6021..6040)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6075..6094
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(6146..7796,1..2))
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     7714..7732
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
ORIGIN
        1 atgggggaca gcggcagcac ctcgcggtag tggctgcgca gcagggagcg cacggctcgg
       61 cagcggggag cgcgcggcat cgcgggggtg gccggggcca gggcttccca cgtgcgcagc
      121 aggacgcagc gctgcctgaa actcgcgccg cgaggagagg gcggggccgc ggaaaggaag
      181 gggaggggct gggagggccc ggagggggct gggccgggga cccgggaggg gtcgggacgg
      241 ggcggggtcc gcgcggagga ggcggagctg gaaggtgaag gggcaggacg ggtgcccggg
      301 tccccagtcc ctccgccacg tgggaagcgc ggtcctgggc gtctgtgccc gcgaatccac
      361 tgggagcccg gcctggcccc gacagcgcag ctgctccggg cggacccggg ggtctgggcc
      421 gcgcttcccc gcccgcgcgc cgctcgcgct cccagggtgc agggacgcca gcgagggccc
      481 cagcggagag aggtcgaatc ggcctaggct gtggggtaac ccgagggagg ggccatgatg
      541 tggaggccct gggaacaggt gcgtgcggcg accctttggc cgctggcctg atccggagac
      601 ccagggctgc ctccaggtcc ggacgcgggg cgtcgggctc cgggcaccac gaatgccgga
      661 cgtgaaggct gagggaccca gaggcagtaa gagaacggtt ttagtcataa atggtaactc
      721 aggggagaga gaaagaaggc tttcagtcac agataactca catgaaagac aaaggttttg
      781 gtcacaaatc ataattcaca cgacagacac taggactcgg agctgaggct gctgcagagt
      841 cagcacaggg tcagggcagg gcccagggcc gctggtgctg gggccttggc taagccgctg
      901 gctgtgcgtc tctggggcag cgccgccctc tagcgggcaa gggaaagggg tcctgcggca
      961 gaaatgccac aacgcgcttc ctatctgggc ctgtcggggt ccacctgtct gtccgcaggc
     1021 ccctgtctgt tccttcctcc cctcagtcca caggcctcct ccttccacct gggttctagg
     1081 acctgggcgg tgtggggcgc ccctcctctc ctgctagaga gccctcagac ccctttttca
     1141 ttcccttggg gggcacaggg acgccctgtg catcccattt gtcctctggc tggcatcact
     1201 gtcctcactc acaggcagca gtgccagcct gaggacttcc atggtggagt gtcgccggat
     1261 cctggatcct agatctggta ccgaggtccc cgggctgcag gatatcctgc agcccgggga
     1321 gctcggtacc agatctgaat tccagctgac tagtgattcg aggtcgacgg tatcgataag
     1381 ctctagaatt attgttcatt tttgagaact cgctcaacga acgatttgat atattttccc
     1441 atttcatcag gtgcatcttc ttgcgaaaaa tgaagacctt ttactttgac aaattcagta
     1501 ttaggaaact tcttggcacc ttcaacaata gcattggaaa agaatcctgg gtccgattca
     1561 ataaacattt ttggtaaatc atcacttgca cgtagataag cattataatt cctaacaatt
     1621 tgtacaacgt caggtttacc accttttact aacgggattt cacgaggcca tgataatgtt
     1681 ggacgacgaa cttcaccttt ctctttgaat ggttcaagat atgctgcaaa ttcttctggt
     1741 tctaactttc tcatgatttt tgatggcaac atggtttcca cgaagaagtt attctccaaa
     1801 accatttttt ctccttcttc agatttgatc aacgcaatat cttcttcaat atcaggccat
     1861 tcatcccatg attcaatcac atctactaca ctttcagcgt gaactattgc tttgatctta
     1921 tcttgatgct catagctata atgaaatgcc aaacaagcac cccaatcatg gccgacaaaa
     1981 atgatcttct ttggtaaatt aagaagttca aaccatgcag taagatattt gtaatgatca
     2041 agtaacctat aagaaccatt accagatttg cctgatttgc ccataccaat aaggtctggt
     2101 ataatacacc gcgctactgg ctcaatatgt ggcacaacat gtcgccataa ataagaagag
     2161 gccgcgttac catgtaaaaa aataacagca ttttctgcat gtttttctga atcataataa
     2221 ttaataaatg aatcaagaac attcatttgt ttacatctgg cccaccactg cggaccagtt
     2281 atcatccgtt tcctttgttc tggatcataa actttcgaag tcatggtggc tagagcttga
     2341 tatcaattca gtaccggaat gccaagccta gcctatagtg agtcgtatta agtactctag
     2401 ccttaagagc tgtaattgaa ctgggagtgg acacctgtgg agagaaaggc aaagtggatg
     2461 tcagtaagac caataggtgc ctatcagaaa cgcaagagtc ttctctgtct cgacaagccc
     2521 agtttctatt ggtctcctta aacctgtctt gtaaccttga tacttacctg cccagtgcct
     2581 cacgaccaac ttcagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt
     2641 ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca
     2701 gcgatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
     2761 gggcgggact atggttgctg actaattgag atgcagatcg cagatctcga gcccgggcta
     2821 gcacgcgtaa gagctcggta cctatcgata gagaaatgtt ctggcacctg cacttgcact
     2881 ggggacagcc tattttgcta gtttgttttg tttcgttttg ttttgatgga gagcgtatgt
     2941 tagtactatc gattcacaca aaaaaccaac acacagatgt aatgaaaata aagatatttt
     3001 attgcggccg ctccaagtac ctcccgtacc ttaatattac ttacttatca tggtagcttg
     3061 ggctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     3121 aatggcgaat ggcaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     3181 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     3241 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     3301 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     3361 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     3421 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     3481 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     3541 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     3601 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3661 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3721 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     3781 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     3841 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     3901 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     3961 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4021 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     4081 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     4141 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     4201 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     4261 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     4321 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     4381 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     4441 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4501 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4561 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     4621 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4681 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4741 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4801 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4861 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4921 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4981 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5041 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5101 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5161 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5221 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5281 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5341 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5401 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     5461 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     5521 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     5581 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     5641 catcggtcga cggatccgct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     5701 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     5761 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5821 accatagtcc cgcccctaac tccgcccagt tccgcccatt ctccgctcca tcgttcagat
     5881 ccttatcgat tttaccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     5941 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     6001 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6061 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgctcgaa
     6121 gcggccggcc gccccgactc tagaattaca cggcgatctt tccgcccttc ttggccttta
     6181 tgaggatctc tctgattttt cttgcgtcga gttttccggt aagacctttc ggtacttcgt
     6241 ccacaaacac aactcctccg cgcaactttt tcgcggttgt tacttgactg gcgacgtaat
     6301 ccacgatctc tttttccgtc atcgtctttc cgtgctccaa aacaacaacg gcggcgggaa
     6361 gttcaccggc gtcatcgtcg ggaagacctg cgacacctgc gtcgaagatg ttggggtgtt
     6421 ggagcaagat ggattccaat tcagcgggag ccacctgata gcctttgtac ttaatcagag
     6481 acttcaggcg gtcaacgatg aagaagtgtt cgtcttcgtc ccagtaagct atgtctccag
     6541 aatgtagcca tccatccttg tcaatcaagg cgttggtcgc ttccggattg tttacataac
     6601 cggacataat cataggacct ctcacacaca gttcgcctct ttgattaacg cccagcgttt
     6661 tcccggtatc cagatccaca accttcgctt caaaaaatgg aacaacttta ccgaccgcgc
     6721 ccggtttatc atccccctcg ggtgtaatca gaatagctga tgtagtctca gtgagcccat
     6781 atccttgcct gatacctggc agatggaacc tcttggcaac cgcttccccg acttccttag
     6841 agaggggagc gccaccagaa gcaatttcgt gtaaattaga taaatcgtat ttgtcaatca
     6901 gagtgctttt ggcgaagaag gagaataggg ttggcaccag cagcgcactt tgaatcttgt
     6961 aatcctgaag gctcctcaga aacagctctt cttcaaatct atacattaag acgactcgaa
     7021 atccacatat caaatatccg agtgtagtaa acattccaaa accgtgatgg aatggaacaa
     7081 cacttaaaat cgcagtatcc ggaatgattt gattgccaaa aataggatct ctggcatgcg
     7141 agaatctcac gcaggcagtt ctatgaggca gagcgacacc tttaggcaga ccagtagatc
     7201 cagaggagtt catgatcagt gcaattgtct tgtccctatc gaaggactct ggcacaaaat
     7261 cgtattcatt aaaaccggga ggtagatgag atgtgacgaa cgtgtacatc gactgaaatc
     7321 cctggtaatc cgttttagaa tccatgataa taattttttg gatgattggg agcttttttt
     7381 gcacgttcaa aattttttgc aacccctttt tggaaacgaa caccacggta ggctgcgaaa
     7441 tgcccatact gttgagcaat tcacgttcat tataaatgtc gttcgcgggc gcaactgcaa
     7501 ctccgataaa taacgcgccc aacaccggca taaagaattg aagagagttt tcactgcata
     7561 cgacgattct gtgatttgta ttcagcccat atcgtttcat agcttctgcc aaccgaacgg
     7621 acatttcgaa gtactcagcg taagtgatgt ccacctcgat atgtgcatct gtaaaagcaa
     7681 ttgttccagg aaccagggcg tatctcttca tagccttatg cagttgctct ccagcggttc
     7741 catcttccag cggatagaat ggcgccgggc ctttctttat gtttttggcg tcttcc
//
