LOCUS       sequence-337236-        7831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly-Renilla luciferase assay for hTERT promoter activity of RE5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7831)
  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 7831)
  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 RE5
FEATURES             Location/Qualifiers
     source          1..7831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             10..1662
                     /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     complement(76..94)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1703..1824
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1714..1733)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1768..1787
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2125..2145
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(2297..2314)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2468..3056)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2548..2567)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3227..4018,4019..4087))
                     /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     3850..3869
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4088..4192)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4219..4674
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4306..4325)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4516..4537
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4805..4853
                     /note="synthetic polyadenylation signal"
     misc_feature    4867..4958
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4907..4926
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        5013..5209
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5060..5195
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          5241..5373
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     5356..5375
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        5418..5436
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5484..6419
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     primer_bind     complement(6433..6449)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6434..6450)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 ctgtccccca tggaagacgc caaaaacata aagaaaggcc cggcgccatt ctatccgctg
       61 gaagatggaa ccgctggaga gcaactgcat aaggctatga agagatacgc cctggttcct
      121 ggaacaattg cttttacaga tgcacatatc gaggtggaca tcacttacgc tgagtacttc
      181 gaaatgtccg ttcggttggc agaagctatg aaacgatatg ggctgaatac aaatcacaga
      241 atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg cgcgttattt
      301 atcggagttg cagttgcgcc cgcgaacgac atttataatg aacgtgaatt gctcaacagt
      361 atgggcattt cgcagcctac cgtggtgttc gtttccaaaa aggggttgca aaaaattttg
      421 aacgtgcaaa aaaagctccc aatcatccaa aaaattatta tcatggattc taaaacggat
      481 taccagggat ttcagtcgat gtacacgttc gtcacatctc atctacctcc cggttttaat
      541 gaatacgatt ttgtgccaga gtccttcgat agggacaaga caattgcact gatcatgaac
      601 tcctctggat ctactggtct gcctaaaggt gtcgctctgc ctcatagaac tgcctgcgtg
      661 agattctcgc atgccagaga tcctattttt ggcaatcaaa tcattccgga tactgcgatt
      721 ttaagtgttg ttccattcca tcacggtttt ggaatgttta ctacactcgg atatttgata
      781 tgtggatttc gagtcgtctt aatgtataga tttgaagaag agctgtttct gaggagcctt
      841 caggattaca agattcaaag tgcgctgctg gtgccaaccc tattctcctt cttcgccaaa
      901 agcactctga ttgacaaata cgatttatct aatttacacg aaattgcttc tggtggcgct
      961 cccctctcta aggaagtcgg ggaagcggtt gccaagaggt tccatctgcc aggtatcagg
     1021 caaggatatg ggctcactga gactacatca gctattctga ttacacccga gggggatgat
     1081 aaaccgggcg cggtcggtaa agttgttcca ttttttgaag cgaaggttgt ggatctggat
     1141 accgggaaaa cgctgggcgt taatcaaaga ggcgaactgt gtgtgagagg tcctatgatt
     1201 atgtccggtt atgtaaacaa tccggaagcg accaacgcct tgattgacaa ggatggatgg
     1261 ctacattctg gagacatagc ttactgggac gaagacgaac acttcttcat cgttgaccgc
     1321 ctgaagtctc tgattaagta caaaggctat caggtggctc ccgctgaatt ggaatccatc
     1381 ttgctccaac accccaacat cttcgacgca ggtgtcgcag gtcttcccga cgatgacgcc
     1441 ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa agacgatgac ggaaaaagag
     1501 atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa agttgcgcgg aggagttgtg
     1561 tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa aatcagagag
     1621 atcctcataa aggccaagaa gggcggaaag atcgccgtgt aattctagag tcggggcggc
     1681 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     1741 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     1801 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     1861 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
     1921 gataaggatc tgaacgatgg agcggagaat gggcggaact gggcggagtt aggggcggga
     1981 ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     2041 gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
     2101 ctggggagcc tggggacttt ccacacccta actgacacac attccacagc ggatccgtcg
     2161 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
     2221 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc
     2281 agcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2341 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2401 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2461 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2521 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2581 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2641 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2701 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2761 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2821 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2881 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     2941 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3001 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3061 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3121 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3181 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3241 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3301 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3361 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3421 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3481 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3541 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3601 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     3661 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     3721 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     3781 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     3841 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     3901 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     3961 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4021 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4081 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4141 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4201 cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4261 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4321 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4381 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4441 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4501 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4561 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4621 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttgccatt
     4681 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     4741 gccagcccaa gctaccatga taagtaagta atattaaggt acgggaggta cttggagcgg
     4801 ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat
     4861 agtactaaca tacgctctcc atcaaaacaa aacgaaacaa aacaaactag caaaataggc
     4921 tgtccccagt gcaagtgcag gtgccagaac atttctctat cgataggtac cgagctctta
     4981 cgcgtgctag cccgggctcg agatctgcga tctgcatctc aattagtcag caaccatagt
     5041 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     5101 ccatcgctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     5161 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctgaagtt
     5221 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     5281 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     5341 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     5401 cttaaggcta gagtacttaa tacgactcac tataggctag gcttggcatt ccggtactga
     5461 attgatatca agctctagcc accatgactt cgaaagttta tgatccagaa caaaggaaac
     5521 ggatgataac tggtccgcag tggtgggcca gatgtaaaca aatgaatgtt cttgattcat
     5581 ttattaatta ttatgattca gaaaaacatg cagaaaatgc tgttattttt ttacatggta
     5641 acgcggcctc ttcttattta tggcgacatg ttgtgccaca tattgagcca gtagcgcggt
     5701 gtattatacc agaccttatt ggtatgggca aatcaggcaa atctggtaat ggttcttata
     5761 ggttacttga tcattacaaa tatcttactg catggtttga acttcttaat ttaccaaaga
     5821 agatcatttt tgtcggccat gattggggtg cttgtttggc atttcattat agctatgagc
     5881 atcaagataa gatcaaagca atagttcacg ctgaaagtgt agtagatgtg attgaatcat
     5941 gggatgaatg gcctgatatt gaagaagata ttgcgttgat caaatctgaa gaaggagaaa
     6001 aaatggtttt ggagaataac ttcttcgtgg aaaccatgtt gccatcaaaa atcatgagaa
     6061 agttagaacc agaagaattt gcagcatatc ttgaaccatt caaagagaaa ggtgaagttc
     6121 gtcgtccaac attatcatgg cctcgtgaaa tcccgttagt aaaaggtggt aaacctgacg
     6181 ttgtacaaat tgttaggaat tataatgctt atctacgtgc aagtgatgat ttaccaaaaa
     6241 tgtttattga atcggaccca ggattctttt ccaatgctat tgttgaaggt gccaagaagt
     6301 ttcctaatac tgaatttgtc aaagtaaaag gtcttcattt ttcgcaagaa gatgcacctg
     6361 atgaaatggg aaaatatatc aaatcgttcg ttgagcgagt tctcaaaaat gaacaataat
     6421 tctagagctt atcgataccg tcgacctcga atcactagtc agctggaatt cagatctggt
     6481 accgagctcc ccgggctgca ggatatcctg cagcccgggg acctcggtac cagatctagg
     6541 atccaggatc cggcgacact ccaccatgga agtcctcagg ggccctggca agtatgtcca
     6601 ctcctctgag tgcagggagg tacagaggtg accagcctga cgtgcagctg tggacacaga
     6661 gggagggctg ctgggggctg gaggccactg cccagtggca gttgtggggg aggcagaaat
     6721 gacctgatcg ctctaaggtg agggagagga gagcccactc tttacagaga cttccccaat
     6781 ggcacaggcg ccacgtggtc ctggggccac tgactgggca gacagcctgg cacagggaca
     6841 ggacaggtct agcctgggat gtgctgagct tgatggatgc atgggaccca ggaggatgtc
     6901 acaatgatga ttcacatggc acacaggtga ccatgactct tccctgatgg agacaatgac
     6961 tcactgctgc agaaaggtgg atctaacccc tccagggtag cctaggagaa caaacagact
     7021 ccactctaag gagcccaggc attccatgct gatccacgcc tgcccagccc ggcggcatcc
     7081 ccactgtctg tgcatggaca ccagccctcc ctggctccag ggcccttgcc ctccagtacc
     7141 ttggcccagc tttggttcta ccccaaggcc tcagccttca cgtccggcat tcgtggtgcc
     7201 cggagcccga cgccccgcgt ccggacctgg aggcagccct gggtctccgg atcaggccag
     7261 cggccaaagg gtcgccgcac gcacctgttc ccagggcctc cacatcatgg cccctccctc
     7321 gggttacccc acagcctagg ccgattcgac ctctctccgc tggggccctc gctggcgtcc
     7381 ctgcaccctg ggagcgcgag cggcgcgcgg gcggggaagc gcggcccaga cccccgggtc
     7441 cgcccggagc agctgcgctg tcggggccag gccgggctcc cagtggattc gcgggcacag
     7501 acgcccagga ccgcgcttcc cacgtggcgg agggactggg gacccgggca cccgtcctgc
     7561 cccttcacct tccagctccg cctcctccgc gcggaccccg ccccgtcccg acccctcccg
     7621 ggtccccggc ccagccccct ccgggccctc ccagcccctc cccttccttt ccgcggcccc
     7681 gccctctcct cgcggcgcga gtttcaggca gcgctgcgtc ctgctgcgca cgtgggaagc
     7741 cctggccccg gccacccccg cgatgccgcg cgctccccgc tgccgagccg tgcgctccct
     7801 gctgcgcagc cactaccgcg aggtgctgcc g
//
