LOCUS       sequence-337243-        7678 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly-Renilla luciferase assay for hTERT promoter activity of RE8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7678)
  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 7678)
  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 RE8
FEATURES             Location/Qualifiers
     source          1..7678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             37..1689
                     /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(103..121)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1730..1851
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1741..1760)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1795..1814
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2152..2172
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(2324..2341)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2495..3083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2575..2594)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3254..4045,4046..4114))
                     /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     3877..3896
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4115..4219)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4246..4701
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4333..4352)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4543..4564
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4832..4880
                     /note="synthetic polyadenylation signal"
     misc_feature    4894..4985
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4934..4953
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        5040..5236
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5087..5222
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          5268..5400
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     5383..5402
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        5445..5463
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5511..6446
                     /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(6460..6476)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6461..6477)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 cgcagccact accgcgaggt gctgccgctg tcccccatgg aagacgccaa aaacataaag
       61 aaaggcccgg cgccattcta tccgctggaa gatggaaccg ctggagagca actgcataag
      121 gctatgaaga gatacgccct ggttcctgga acaattgctt ttacagatgc acatatcgag
      181 gtggacatca cttacgctga gtacttcgaa atgtccgttc ggttggcaga agctatgaaa
      241 cgatatgggc tgaatacaaa tcacagaatc gtcgtatgca gtgaaaactc tcttcaattc
      301 tttatgccgg tgttgggcgc gttatttatc ggagttgcag ttgcgcccgc gaacgacatt
      361 tataatgaac gtgaattgct caacagtatg ggcatttcgc agcctaccgt ggtgttcgtt
      421 tccaaaaagg ggttgcaaaa aattttgaac gtgcaaaaaa agctcccaat catccaaaaa
      481 attattatca tggattctaa aacggattac cagggatttc agtcgatgta cacgttcgtc
      541 acatctcatc tacctcccgg ttttaatgaa tacgattttg tgccagagtc cttcgatagg
      601 gacaagacaa ttgcactgat catgaactcc tctggatcta ctggtctgcc taaaggtgtc
      661 gctctgcctc atagaactgc ctgcgtgaga ttctcgcatg ccagagatcc tatttttggc
      721 aatcaaatca ttccggatac tgcgatttta agtgttgttc cattccatca cggttttgga
      781 atgtttacta cactcggata tttgatatgt ggatttcgag tcgtcttaat gtatagattt
      841 gaagaagagc tgtttctgag gagccttcag gattacaaga ttcaaagtgc gctgctggtg
      901 ccaaccctat tctccttctt cgccaaaagc actctgattg acaaatacga tttatctaat
      961 ttacacgaaa ttgcttctgg tggcgctccc ctctctaagg aagtcgggga agcggttgcc
     1021 aagaggttcc atctgccagg tatcaggcaa ggatatgggc tcactgagac tacatcagct
     1081 attctgatta cacccgaggg ggatgataaa ccgggcgcgg tcggtaaagt tgttccattt
     1141 tttgaagcga aggttgtgga tctggatacc gggaaaacgc tgggcgttaa tcaaagaggc
     1201 gaactgtgtg tgagaggtcc tatgattatg tccggttatg taaacaatcc ggaagcgacc
     1261 aacgccttga ttgacaagga tggatggcta cattctggag acatagctta ctgggacgaa
     1321 gacgaacact tcttcatcgt tgaccgcctg aagtctctga ttaagtacaa aggctatcag
     1381 gtggctcccg ctgaattgga atccatcttg ctccaacacc ccaacatctt cgacgcaggt
     1441 gtcgcaggtc ttcccgacga tgacgccggt gaacttcccg ccgccgttgt tgttttggag
     1501 cacggaaaga cgatgacgga aaaagagatc gtggattacg tcgccagtca agtaacaacc
     1561 gcgaaaaagt tgcgcggagg agttgtgttt gtggacgaag taccgaaagg tcttaccgga
     1621 aaactcgacg caagaaaaat cagagagatc ctcataaagg ccaagaaggg cggaaagatc
     1681 gccgtgtaat tctagagtcg gggcggccgg ccgcttcgag cagacatgat aagatacatt
     1741 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     1801 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     1861 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     1921 taaaacctct acaaatgtgg taaaatcgat aaggatctga acgatggagc ggagaatggg
     1981 cggaactggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt
     2041 gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga
     2101 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact
     2161 gacacacatt ccacagcgga tccgtcgacc gatgcccttg agagccttca acccagtcag
     2221 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
     2281 catgcaactc gtaggacagg tgccggcagc gctcttccgc ttcctcgctc actgactcgc
     2341 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2401 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2461 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2521 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2581 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2641 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2701 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2761 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2821 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2881 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2941 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3001 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3061 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3121 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3181 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3241 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3301 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3361 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3421 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3481 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3541 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3601 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3661 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3721 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3781 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     3841 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     3901 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     3961 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4021 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4081 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4141 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4201 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg
     4261 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4321 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4381 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4441 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4501 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4561 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     4621 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     4681 aaatattaac gcttacaatt tgccattcgc cattcaggct gcgcaactgt tgggaagggc
     4741 gatcggtgcg ggcctcttcg ctattacgcc agcccaagct accatgataa gtaagtaata
     4801 ttaaggtacg ggaggtactt ggagcggccg caataaaata tctttatttt cattacatct
     4861 gtgtgttggt tttttgtgtg aatcgatagt actaacatac gctctccatc aaaacaaaac
     4921 gaaacaaaac aaactagcaa aataggctgt ccccagtgca agtgcaggtg ccagaacatt
     4981 tctctatcga taggtaccga gctcttacgc gtgctagccc gggctcgaga tctgcgatct
     5041 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     5101 tccgcccagt tccgcccatt ctccgcccca tcgctgacta atttttttta tttatgcaga
     5161 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     5221 cctaggcttt tgcaaaaagc tgaagttggt cgtgaggcac tgggcaggta agtatcaagg
     5281 ttacaagaca ggtttaagga gaccaataga aactgggctt gtcgagacag agaagactct
     5341 tgcgtttctg ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag
     5401 gtgtccactc ccagttcaat tacagctctt aaggctagag tacttaatac gactcactat
     5461 aggctaggct tggcattccg gtactgaatt gatatcaagc tctagccacc atgacttcga
     5521 aagtttatga tccagaacaa aggaaacgga tgataactgg tccgcagtgg tgggccagat
     5581 gtaaacaaat gaatgttctt gattcattta ttaattatta tgattcagaa aaacatgcag
     5641 aaaatgctgt tattttttta catggtaacg cggcctcttc ttatttatgg cgacatgttg
     5701 tgccacatat tgagccagta gcgcggtgta ttataccaga ccttattggt atgggcaaat
     5761 caggcaaatc tggtaatggt tcttataggt tacttgatca ttacaaatat cttactgcat
     5821 ggtttgaact tcttaattta ccaaagaaga tcatttttgt cggccatgat tggggtgctt
     5881 gtttggcatt tcattatagc tatgagcatc aagataagat caaagcaata gttcacgctg
     5941 aaagtgtagt agatgtgatt gaatcatggg atgaatggcc tgatattgaa gaagatattg
     6001 cgttgatcaa atctgaagaa ggagaaaaaa tggttttgga gaataacttc ttcgtggaaa
     6061 ccatgttgcc atcaaaaatc atgagaaagt tagaaccaga agaatttgca gcatatcttg
     6121 aaccattcaa agagaaaggt gaagttcgtc gtccaacatt atcatggcct cgtgaaatcc
     6181 cgttagtaaa aggtggtaaa cctgacgttg tacaaattgt taggaattat aatgcttatc
     6241 tacgtgcaag tgatgattta ccaaaaatgt ttattgaatc ggacccagga ttcttttcca
     6301 atgctattgt tgaaggtgcc aagaagtttc ctaatactga atttgtcaaa gtaaaaggtc
     6361 ttcatttttc gcaagaagat gcacctgatg aaatgggaaa atatatcaaa tcgttcgttg
     6421 agcgagttct caaaaatgaa caataattct agagcttatc gataccgtcg acctcgaatc
     6481 actagtcagc tggaattcag atctggtacc gagctccccg ggctgcagga tatcctgcag
     6541 cccggggacc tcggtaccag atctaggatc caggatccgg cgacactcca ccatggaagt
     6601 cctcagggcc ttggacagct tgcagaactc actcagagca cctaagaagg atgtaaatcc
     6661 ctcccttgcc gccaggtgcc tggtggaggg gagtgatggg tatagaggtg tcggggagag
     6721 cgcggggtgt ggtgctgagt ggtctctcag tgggctaaga gctcagggct gcccgaggtg
     6781 cagggacccc cggggcagga ggagtcatag ccgctgtgtg tgtgatgtat gcagtgggtc
     6841 ttgaggcagg ggtggcatcc gccaaacggg gagggagggc tacgctatga taaagagttc
     6901 gaggtttatt tttcagataa atgtttggga tttggggcat atcttataaa tgttcacaca
     6961 gaaaccaacg tggttctgtt tttttaaaca ttcaagtatg cgagaacaag aagtttaggt
     7021 gtgaattcct caaattctga gccctcagcc ttcacgtccg gcattcgtgg tgcccggagc
     7081 ccgacgcccc gcgtccggac ctggaggcag ccctgggtct ccggatcagg ccagcggcca
     7141 aagggtcgcc gcacgcacct gttcccaggg cctccacatc atggcccctc cctcgggtta
     7201 ccccacagcc taggccgatt cgacctctct ccgctggggc cctcgctggc gtccctgcac
     7261 cctgggagcg cgagcggcgc gcgggcgggg aagcgcggcc cagacccccg ggtccgcccg
     7321 gagcagctgc gctgtcgggg ccaggccggg ctcccagtgg attcgcgggc acagacgccc
     7381 aggaccgcgc ttcccacgtg gcggagggac tggggacccg ggcacccgtc ctgccccttc
     7441 accttccagc tccgcctcct ccgcgcggac cccgccccgt cccgacccct cccgggtccc
     7501 cggcccagcc ccctccgggc cctcccagcc cctccccttc ctttccgcgg ccccgccctc
     7561 tcctcgcggc gcgagtttca ggcagcgctg cgtcctgctg cgcacgtggg aagccctggc
     7621 cccggccacc cccgcgatgc cgcgcgctcc ccgctgccga gccgtgcgct ccctgctg
//
