LOCUS       sequence-337241-        7853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly-Renilla luciferase assay for hTERT promoter activity of RE7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7853)
  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 7853)
  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 RE7
FEATURES             Location/Qualifiers
     source          1..7853
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             714..2366
                     /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(780..798)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2407..2528
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2418..2437)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2472..2491
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2829..2849
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(3001..3018)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3172..3760)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3252..3271)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3931..4722,4723..4791))
                     /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     4554..4573
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4792..4896)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4923..5378
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5010..5029)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5220..5241
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5509..5557
                     /note="synthetic polyadenylation signal"
     misc_feature    5571..5662
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5611..5630
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        5717..5913
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5764..5899
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          5945..6077
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     6060..6079
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        6122..6140
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6188..7123
                     /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(7137..7153)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7138..7154)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 gcccaaaccc tcaaaagctg caggagccca caggctcgag ccctcagcct tcacgtccgg
       61 cattcgtggt gcccggagcc cgacgccccg cgtccggacc tggaggcagc cctgggtctc
      121 cggatcaggc cagcggccaa agggtcgccg cacgcacctg ttcccagggc ctccacatca
      181 tggcccctcc ctcgggttac cccacagcct aggccgattc gacctctctc cgctggggcc
      241 ctcgctggcg tccctgcacc ctgggagcgc gagcggcgcg cgggcgggga agcgcggccc
      301 agacccccgg gtccgcccgg agcagctgcg ctgtcggggc caggccgggc tcccagtgga
      361 ttcgcgggca cagacgccca ggaccgcgct tcccacgtgg cggagggact ggggacccgg
      421 gcacccgtcc tgccccttca ccttccagct ccgcctcctc cgcgcggacc ccgccccgtc
      481 ccgacccctc ccgggtcccc ggcccagccc cctccgggcc ctcccagccc ctccccttcc
      541 tttccgcggc cccgccctct cctcgcggcg cgagtttcag gcagcgctgc gtcctgctgc
      601 gcacgtggga agccctggcc ccggccaccc ccgcgatgcc gcgcgctccc cgctgccgag
      661 ccgtgcgctc cctgctgcgc agccactacc gcgaggtgct gccgctgtcc cccatggaag
      721 acgccaaaaa cataaagaaa ggcccggcgc cattctatcc gctggaagat ggaaccgctg
      781 gagagcaact gcataaggct atgaagagat acgccctggt tcctggaaca attgctttta
      841 cagatgcaca tatcgaggtg gacatcactt acgctgagta cttcgaaatg tccgttcggt
      901 tggcagaagc tatgaaacga tatgggctga atacaaatca cagaatcgtc gtatgcagtg
      961 aaaactctct tcaattcttt atgccggtgt tgggcgcgtt atttatcgga gttgcagttg
     1021 cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa cagtatgggc atttcgcagc
     1081 ctaccgtggt gttcgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaagc
     1141 tcccaatcat ccaaaaaatt attatcatgg attctaaaac ggattaccag ggatttcagt
     1201 cgatgtacac gttcgtcaca tctcatctac ctcccggttt taatgaatac gattttgtgc
     1261 cagagtcctt cgatagggac aagacaattg cactgatcat gaactcctct ggatctactg
     1321 gtctgcctaa aggtgtcgct ctgcctcata gaactgcctg cgtgagattc tcgcatgcca
     1381 gagatcctat ttttggcaat caaatcattc cggatactgc gattttaagt gttgttccat
     1441 tccatcacgg ttttggaatg tttactacac tcggatattt gatatgtgga tttcgagtcg
     1501 tcttaatgta tagatttgaa gaagagctgt ttctgaggag ccttcaggat tacaagattc
     1561 aaagtgcgct gctggtgcca accctattct ccttcttcgc caaaagcact ctgattgaca
     1621 aatacgattt atctaattta cacgaaattg cttctggtgg cgctcccctc tctaaggaag
     1681 tcggggaagc ggttgccaag aggttccatc tgccaggtat caggcaagga tatgggctca
     1741 ctgagactac atcagctatt ctgattacac ccgaggggga tgataaaccg ggcgcggtcg
     1801 gtaaagttgt tccatttttt gaagcgaagg ttgtggatct ggataccggg aaaacgctgg
     1861 gcgttaatca aagaggcgaa ctgtgtgtga gaggtcctat gattatgtcc ggttatgtaa
     1921 acaatccgga agcgaccaac gccttgattg acaaggatgg atggctacat tctggagaca
     1981 tagcttactg ggacgaagac gaacacttct tcatcgttga ccgcctgaag tctctgatta
     2041 agtacaaagg ctatcaggtg gctcccgctg aattggaatc catcttgctc caacacccca
     2101 acatcttcga cgcaggtgtc gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg
     2161 ccgttgttgt tttggagcac ggaaagacga tgacggaaaa agagatcgtg gattacgtcg
     2221 ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac
     2281 cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca
     2341 agaagggcgg aaagatcgcc gtgtaattct agagtcgggg cggccggccg cttcgagcag
     2401 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     2461 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     2521 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     2581 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatctgaacg
     2641 atggagcgga gaatgggcgg aactgggcgg agttaggggc gggactatgg ttgctgacta
     2701 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
     2761 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     2821 ctttccacac cctaactgac acacattcca cagcggatcc gtcgaccgat gcccttgaga
     2881 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     2941 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc
     3001 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3061 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3121 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3181 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3241 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3301 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3361 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3421 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3481 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3541 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3601 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3661 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3721 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3781 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3841 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3901 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3961 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4021 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4081 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4141 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4201 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4261 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4321 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4381 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4441 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4501 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4561 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4621 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4681 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4741 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4801 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4861 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4921 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4981 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5041 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     5101 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5161 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5221 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5281 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5341 taacgcgaat tttaacaaaa tattaacgct tacaatttgc cattcgccat tcaggctgcg
     5401 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc
     5461 atgataagta agtaatatta aggtacggga ggtacttgga gcggccgcaa taaaatatct
     5521 ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct
     5581 ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt
     5641 gcaggtgcca gaacatttct ctatcgatag gtaccgagct cttacgcgtg ctagcccggg
     5701 ctcgagatct gcgatctgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5761 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatcg ctgactaatt
     5821 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     5881 ggaggctttt ttggaggcct aggcttttgc aaaaagctga agttggtcgt gaggcactgg
     5941 gcaggtaagt atcaaggtta caagacaggt ttaaggagac caatagaaac tgggcttgtc
     6001 gagacagaga agactcttgc gtttctgata ggcacctatt ggtcttactg acatccactt
     6061 tgcctttctc tccacaggtg tccactccca gttcaattac agctcttaag gctagagtac
     6121 ttaatacgac tcactatagg ctaggcttgg cattccggta ctgaattgat atcaagctct
     6181 agccaccatg acttcgaaag tttatgatcc agaacaaagg aaacggatga taactggtcc
     6241 gcagtggtgg gccagatgta aacaaatgaa tgttcttgat tcatttatta attattatga
     6301 ttcagaaaaa catgcagaaa atgctgttat ttttttacat ggtaacgcgg cctcttctta
     6361 tttatggcga catgttgtgc cacatattga gccagtagcg cggtgtatta taccagacct
     6421 tattggtatg ggcaaatcag gcaaatctgg taatggttct tataggttac ttgatcatta
     6481 caaatatctt actgcatggt ttgaacttct taatttacca aagaagatca tttttgtcgg
     6541 ccatgattgg ggtgcttgtt tggcatttca ttatagctat gagcatcaag ataagatcaa
     6601 agcaatagtt cacgctgaaa gtgtagtaga tgtgattgaa tcatgggatg aatggcctga
     6661 tattgaagaa gatattgcgt tgatcaaatc tgaagaagga gaaaaaatgg ttttggagaa
     6721 taacttcttc gtggaaacca tgttgccatc aaaaatcatg agaaagttag aaccagaaga
     6781 atttgcagca tatcttgaac cattcaaaga gaaaggtgaa gttcgtcgtc caacattatc
     6841 atggcctcgt gaaatcccgt tagtaaaagg tggtaaacct gacgttgtac aaattgttag
     6901 gaattataat gcttatctac gtgcaagtga tgatttacca aaaatgttta ttgaatcgga
     6961 cccaggattc ttttccaatg ctattgttga aggtgccaag aagtttccta atactgaatt
     7021 tgtcaaagta aaaggtcttc atttttcgca agaagatgca cctgatgaaa tgggaaaata
     7081 tatcaaatcg ttcgttgagc gagttctcaa aaatgaacaa taattctaga gcttatcgat
     7141 accgtcgacc tcgaatcact agtcagctgg aattcagatc tggtaccgag ctccccgggc
     7201 tgcaggatat cctgcagccc ggggacctcg gtaccagatc taggatccag gatccggcga
     7261 cactccacca tggaagtcct cagggctcga gcccacctcc agccaaaaca ggaccttcac
     7321 acataccagc acgcttccca ttcagcccag gccctgagac aggccctccg gagctgtggc
     7381 ccagggaaac gccaatctcc ccggacctcc cgcctggtct gagtaaaaag aatcctgaaa
     7441 ttcaggatcc gtggcatttc tagctgcttg catttgttta acaaacatct gcagaaagcc
     7501 agtggggccc tggtggagag cacctcccaa gccccctgtg gtgggagccc cacccccagc
     7561 acatcctgca tcaggggatc ctgctacaag aaagagtccc cagcctgctg gcccttccta
     7621 ctccgaactg gccgcagata cagggagccc cccagcacat cctgcatcag gggatcctgc
     7681 tacaagaaag agtccccagc ctgccggccc ttcctactcc gaactggccg cagatacagg
     7741 gagcccccca gcacatcctg catcagggga tcctgctaca agagtcccca gcctgcccgg
     7801 cccttcctac tccgaactgg ctgcagatac agggagcccc ccagcacatc ctg
//
