LOCUS       sequence-337237-        7719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly-Renilla luciferase assay for hTERT promoter activity of RE4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7719)
  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 7719)
  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 RE4
FEATURES             Location/Qualifiers
     source          1..7719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             382..2034
                     /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(448..466)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2075..2196
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2086..2105)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2140..2159
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2497..2517
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(2669..2686)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2840..3428)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2920..2939)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3599..4390,4391..4459))
                     /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     4222..4241
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4460..4564)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4591..5046
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4678..4697)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4888..4909
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5177..5225
                     /note="synthetic polyadenylation signal"
     misc_feature    5239..5330
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5279..5298
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        5385..5581
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5432..5567
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          5613..5745
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     5728..5747
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        5790..5808
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5856..6791
                     /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(6805..6821)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6806..6822)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 gtcggggcca ggccgggctc ccagtggatt cgcgggcaca gacgcccagg accgcgcttc
       61 ccacgtggcg gagggactgg ggacccgggc acccgtcctg ccccttcacc ttccagctcc
      121 gcctcctccg cgcggacccc gccccgtccc gacccctccc gggtccccgg cccagccccc
      181 tccgggccct cccagcccct ccccttcctt tccgcggccc cgccctctcc tcgcggcgcg
      241 agtttcaggc agcgctgcgt cctgctgcgc acgtgggaag ccctggcccc ggccaccccc
      301 gcgatgccgc gcgctccccg ctgccgagcc gtgcgctccc tgctgcgcag ccactaccgc
      361 gaggtgctgc cgctgtcccc catggaagac gccaaaaaca taaagaaagg cccggcgcca
      421 ttctatccgc tggaagatgg aaccgctgga gagcaactgc ataaggctat gaagagatac
      481 gccctggttc ctggaacaat tgcttttaca gatgcacata tcgaggtgga catcacttac
      541 gctgagtact tcgaaatgtc cgttcggttg gcagaagcta tgaaacgata tgggctgaat
      601 acaaatcaca gaatcgtcgt atgcagtgaa aactctcttc aattctttat gccggtgttg
      661 ggcgcgttat ttatcggagt tgcagttgcg cccgcgaacg acatttataa tgaacgtgaa
      721 ttgctcaaca gtatgggcat ttcgcagcct accgtggtgt tcgtttccaa aaaggggttg
      781 caaaaaattt tgaacgtgca aaaaaagctc ccaatcatcc aaaaaattat tatcatggat
      841 tctaaaacgg attaccaggg atttcagtcg atgtacacgt tcgtcacatc tcatctacct
      901 cccggtttta atgaatacga ttttgtgcca gagtccttcg atagggacaa gacaattgca
      961 ctgatcatga actcctctgg atctactggt ctgcctaaag gtgtcgctct gcctcataga
     1021 actgcctgcg tgagattctc gcatgccaga gatcctattt ttggcaatca aatcattccg
     1081 gatactgcga ttttaagtgt tgttccattc catcacggtt ttggaatgtt tactacactc
     1141 ggatatttga tatgtggatt tcgagtcgtc ttaatgtata gatttgaaga agagctgttt
     1201 ctgaggagcc ttcaggatta caagattcaa agtgcgctgc tggtgccaac cctattctcc
     1261 ttcttcgcca aaagcactct gattgacaaa tacgatttat ctaatttaca cgaaattgct
     1321 tctggtggcg ctcccctctc taaggaagtc ggggaagcgg ttgccaagag gttccatctg
     1381 ccaggtatca ggcaaggata tgggctcact gagactacat cagctattct gattacaccc
     1441 gagggggatg ataaaccggg cgcggtcggt aaagttgttc cattttttga agcgaaggtt
     1501 gtggatctgg ataccgggaa aacgctgggc gttaatcaaa gaggcgaact gtgtgtgaga
     1561 ggtcctatga ttatgtccgg ttatgtaaac aatccggaag cgaccaacgc cttgattgac
     1621 aaggatggat ggctacattc tggagacata gcttactggg acgaagacga acacttcttc
     1681 atcgttgacc gcctgaagtc tctgattaag tacaaaggct atcaggtggc tcccgctgaa
     1741 ttggaatcca tcttgctcca acaccccaac atcttcgacg caggtgtcgc aggtcttccc
     1801 gacgatgacg ccggtgaact tcccgccgcc gttgttgttt tggagcacgg aaagacgatg
     1861 acggaaaaag agatcgtgga ttacgtcgcc agtcaagtaa caaccgcgaa aaagttgcgc
     1921 ggaggagttg tgtttgtgga cgaagtaccg aaaggtctta ccggaaaact cgacgcaaga
     1981 aaaatcagag agatcctcat aaaggccaag aagggcggaa agatcgccgt gtaattctag
     2041 agtcggggcg gccggccgct tcgagcagac atgataagat acattgatga gtttggacaa
     2101 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     2161 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     2221 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     2281 tgtggtaaaa tcgataagga tctgaacgat ggagcggaga atgggcggaa ctgggcggag
     2341 ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
     2401 ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca
     2461 tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac acattccaca
     2521 gcggatccgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc
     2581 gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg
     2641 acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2701 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2761 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2821 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2881 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2941 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3001 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3061 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3121 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3181 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3241 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3301 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3361 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3421 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3481 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3541 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3601 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3661 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3721 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3781 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3841 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3901 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3961 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4021 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4081 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4141 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4201 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4261 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4321 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4381 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4441 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4501 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4561 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
     4621 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4681 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4741 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4801 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4861 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4921 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4981 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     5041 caatttgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     5101 cttcgctatt acgccagccc aagctaccat gataagtaag taatattaag gtacgggagg
     5161 tacttggagc ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     5221 gtgtgaatcg atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact
     5281 agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct atcgataggt
     5341 accgagctct tacgcgtgct agcccgggct cgagatctgc gatctgcatc tcaattagtc
     5401 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     5461 ccattctccg ccccatcgct gactaatttt ttttatttat gcagaggccg aggccgcctc
     5521 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     5581 aaagctgaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt
     5641 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
     5701 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt
     5761 tcaattacag ctcttaaggc tagagtactt aatacgactc actataggct aggcttggca
     5821 ttccggtact gaattgatat caagctctag ccaccatgac ttcgaaagtt tatgatccag
     5881 aacaaaggaa acggatgata actggtccgc agtggtgggc cagatgtaaa caaatgaatg
     5941 ttcttgattc atttattaat tattatgatt cagaaaaaca tgcagaaaat gctgttattt
     6001 ttttacatgg taacgcggcc tcttcttatt tatggcgaca tgttgtgcca catattgagc
     6061 cagtagcgcg gtgtattata ccagacctta ttggtatggg caaatcaggc aaatctggta
     6121 atggttctta taggttactt gatcattaca aatatcttac tgcatggttt gaacttctta
     6181 atttaccaaa gaagatcatt tttgtcggcc atgattgggg tgcttgtttg gcatttcatt
     6241 atagctatga gcatcaagat aagatcaaag caatagttca cgctgaaagt gtagtagatg
     6301 tgattgaatc atgggatgaa tggcctgata ttgaagaaga tattgcgttg atcaaatctg
     6361 aagaaggaga aaaaatggtt ttggagaata acttcttcgt ggaaaccatg ttgccatcaa
     6421 aaatcatgag aaagttagaa ccagaagaat ttgcagcata tcttgaacca ttcaaagaga
     6481 aaggtgaagt tcgtcgtcca acattatcat ggcctcgtga aatcccgtta gtaaaaggtg
     6541 gtaaacctga cgttgtacaa attgttagga attataatgc ttatctacgt gcaagtgatg
     6601 atttaccaaa aatgtttatt gaatcggacc caggattctt ttccaatgct attgttgaag
     6661 gtgccaagaa gtttcctaat actgaatttg tcaaagtaaa aggtcttcat ttttcgcaag
     6721 aagatgcacc tgatgaaatg ggaaaatata tcaaatcgtt cgttgagcga gttctcaaaa
     6781 atgaacaata attctagagc ttatcgatac cgtcgacctc gaatcactag tcagctggaa
     6841 ttcagatctg gtaccgagct ccccgggctg caggatatcc tgcagcccgg ggacctcggt
     6901 accagatcta ggatccagga tccggcgaca ctccaccatg gaagtcctca ggtccacgga
     6961 ggtcacgttc aggctaagct gggcagcccg gccttgtgtt gctggccccc agccgaggga
     7021 ggagctacgg gaggggttgg ggtgaggctg tggctgggat gtgcagtgcc cagcgacgga
     7081 gggtgctgga catgtgcccg gctactggac agcagccagg gagcaagtca caacagaagc
     7141 cgagggggag gatcctgcat ccccactgct ccagaaggtg catccagctg gggcgccctc
     7201 accgtgcgga cacaggggct cccagctttc tgtgcacgca aggccacagc aggagcttcc
     7261 cggggtctgc agatgaccca aagcagagtt cattttcttc cagtaccctc tctgcttcta
     7321 gaaccttcct ccagccccat tccagcgctc actctggccg agacccctgc ccactcactg
     7381 gttggtggcc ggccagccgc ccaccccacg tcagcaaagc cagctgcccc tcagccttca
     7441 cgtccggcat tcgtggtgcc cggagcccga cgccccgcgt ccggacctgg aggcagccct
     7501 gggtctccgg atcaggccag cggccaaagg gtcgccgcac gcacctgttc ccagggcctc
     7561 cacatcatgg cccctccctc gggttacccc acagcctagg ccgattcgac ctctctccgc
     7621 tggggccctc gctggcgtcc ctgcaccctg ggagcgcgag cggcgcgcgg gcggggaagc
     7681 gcggcccaga cccccgggtc cgcccggagc agctgcgct
//
