LOCUS       sequence-337041-        7607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly-Renilla luciferase assay for hTERT promoter activity of RE1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7607)
  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 7607)
  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 RE1
FEATURES             Location/Qualifiers
     source          1..7607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             53..1705
                     /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(119..137)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1746..1867
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1757..1776)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1811..1830
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2168..2188
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(2340..2357)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2511..3099)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2591..2610)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3270..4061,4062..4130))
                     /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     3893..3912
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4131..4235)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4262..4717
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4349..4368)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4559..4580
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4848..4896
                     /note="synthetic polyadenylation signal"
     misc_feature    4910..5001
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4950..4969
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        5056..5252
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5103..5238
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          5284..5416
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     5399..5418
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        5461..5479
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5527..6462
                     /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(6476..6492)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6477..6493)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 cgtgcgctcc ctgctgcgca gccactaccg cgaggtgctg ccgctgtccc ccatggaaga
       61 cgccaaaaac ataaagaaag gcccggcgcc attctatccg ctggaagatg gaaccgctgg
      121 agagcaactg cataaggcta tgaagagata cgccctggtt cctggaacaa ttgcttttac
      181 agatgcacat atcgaggtgg acatcactta cgctgagtac ttcgaaatgt ccgttcggtt
      241 ggcagaagct atgaaacgat atgggctgaa tacaaatcac agaatcgtcg tatgcagtga
      301 aaactctctt caattcttta tgccggtgtt gggcgcgtta tttatcggag ttgcagttgc
      361 gcccgcgaac gacatttata atgaacgtga attgctcaac agtatgggca tttcgcagcc
      421 taccgtggtg ttcgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc aaaaaaagct
      481 cccaatcatc caaaaaatta ttatcatgga ttctaaaacg gattaccagg gatttcagtc
      541 gatgtacacg ttcgtcacat ctcatctacc tcccggtttt aatgaatacg attttgtgcc
      601 agagtccttc gatagggaca agacaattgc actgatcatg aactcctctg gatctactgg
      661 tctgcctaaa ggtgtcgctc tgcctcatag aactgcctgc gtgagattct cgcatgccag
      721 agatcctatt tttggcaatc aaatcattcc ggatactgcg attttaagtg ttgttccatt
      781 ccatcacggt tttggaatgt ttactacact cggatatttg atatgtggat ttcgagtcgt
      841 cttaatgtat agatttgaag aagagctgtt tctgaggagc cttcaggatt acaagattca
      901 aagtgcgctg ctggtgccaa ccctattctc cttcttcgcc aaaagcactc tgattgacaa
      961 atacgattta tctaatttac acgaaattgc ttctggtggc gctcccctct ctaaggaagt
     1021 cggggaagcg gttgccaaga ggttccatct gccaggtatc aggcaaggat atgggctcac
     1081 tgagactaca tcagctattc tgattacacc cgagggggat gataaaccgg gcgcggtcgg
     1141 taaagttgtt ccattttttg aagcgaaggt tgtggatctg gataccggga aaacgctggg
     1201 cgttaatcaa agaggcgaac tgtgtgtgag aggtcctatg attatgtccg gttatgtaaa
     1261 caatccggaa gcgaccaacg ccttgattga caaggatgga tggctacatt ctggagacat
     1321 agcttactgg gacgaagacg aacacttctt catcgttgac cgcctgaagt ctctgattaa
     1381 gtacaaaggc tatcaggtgg ctcccgctga attggaatcc atcttgctcc aacaccccaa
     1441 catcttcgac gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac ttcccgccgc
     1501 cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg attacgtcgc
     1561 cagtcaagta acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg acgaagtacc
     1621 gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga gagatcctca taaaggccaa
     1681 gaagggcgga aagatcgccg tgtaattcta gagtcggggc ggccggccgc ttcgagcaga
     1741 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     1801 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     1861 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     1921 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg atctgaacga
     1981 tggagcggag aatgggcgga actgggcgga gttaggggcg ggactatggt tgctgactaa
     2041 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
     2101 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
     2161 tttccacacc ctaactgaca cacattccac agcggatccg tcgaccgatg cccttgagag
     2221 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta
     2281 tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc
     2341 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2401 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2461 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2521 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2581 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2641 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2701 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2761 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2821 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2881 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2941 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3001 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3061 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3121 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3181 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3241 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3301 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3361 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3421 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3481 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3541 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3601 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3661 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3721 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3781 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3841 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3901 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3961 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4021 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4081 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4141 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4201 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4261 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4321 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4381 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4441 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4501 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4561 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4621 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4681 aacgcgaatt ttaacaaaat attaacgctt acaatttgcc attcgccatt caggctgcgc
     4741 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagcc caagctacca
     4801 tgataagtaa gtaatattaa ggtacgggag gtacttggag cggccgcaat aaaatatctt
     4861 tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagtacta acatacgctc
     4921 tccatcaaaa caaaacgaaa caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg
     4981 caggtgccag aacatttctc tatcgatagg taccgagctc ttacgcgtgc tagcccgggc
     5041 tcgagatctg cgatctgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     5101 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatcgc tgactaattt
     5161 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     5221 gaggcttttt tggaggccta ggcttttgca aaaagctgaa gttggtcgtg aggcactggg
     5281 caggtaagta tcaaggttac aagacaggtt taaggagacc aatagaaact gggcttgtcg
     5341 agacagagaa gactcttgcg tttctgatag gcacctattg gtcttactga catccacttt
     5401 gcctttctct ccacaggtgt ccactcccag ttcaattaca gctcttaagg ctagagtact
     5461 taatacgact cactataggc taggcttggc attccggtac tgaattgata tcaagctcta
     5521 gccaccatga cttcgaaagt ttatgatcca gaacaaagga aacggatgat aactggtccg
     5581 cagtggtggg ccagatgtaa acaaatgaat gttcttgatt catttattaa ttattatgat
     5641 tcagaaaaac atgcagaaaa tgctgttatt tttttacatg gtaacgcggc ctcttcttat
     5701 ttatggcgac atgttgtgcc acatattgag ccagtagcgc ggtgtattat accagacctt
     5761 attggtatgg gcaaatcagg caaatctggt aatggttctt ataggttact tgatcattac
     5821 aaatatctta ctgcatggtt tgaacttctt aatttaccaa agaagatcat ttttgtcggc
     5881 catgattggg gtgcttgttt ggcatttcat tatagctatg agcatcaaga taagatcaaa
     5941 gcaatagttc acgctgaaag tgtagtagat gtgattgaat catgggatga atggcctgat
     6001 attgaagaag atattgcgtt gatcaaatct gaagaaggag aaaaaatggt tttggagaat
     6061 aacttcttcg tggaaaccat gttgccatca aaaatcatga gaaagttaga accagaagaa
     6121 tttgcagcat atcttgaacc attcaaagag aaaggtgaag ttcgtcgtcc aacattatca
     6181 tggcctcgtg aaatcccgtt agtaaaaggt ggtaaacctg acgttgtaca aattgttagg
     6241 aattataatg cttatctacg tgcaagtgat gatttaccaa aaatgtttat tgaatcggac
     6301 ccaggattct tttccaatgc tattgttgaa ggtgccaaga agtttcctaa tactgaattt
     6361 gtcaaagtaa aaggtcttca tttttcgcaa gaagatgcac ctgatgaaat gggaaaatat
     6421 atcaaatcgt tcgttgagcg agttctcaaa aatgaacaat aattctagag cttatcgata
     6481 ccgtcgacct cgaatcacta gtcagctgga attcagatct ggtaccgagc tccccgggct
     6541 gcaggatatc ctgcagcccg gggacctcgg taccagatct aggatccagg atccggcgac
     6601 actccaccat ggaagtcctc aggagtaggc gggatccgga caggtctgcc caatcccctc
     6661 gcagttctca ggggagggac agaactaagg gaaaccccct ccgagatcct cctcatgcca
     6721 ggctccctcc tcacagagac cctggcaggg tctgctgagg aacagcacac ccaggatcct
     6781 ggaactgagt ctgcgggcct cagcggttcg cgttttgcta ccaacggaac accccctccc
     6841 ctcctccagc cacgcaggcg cactctgcct gggtcctggg accaggctcc ccacgggctc
     6901 ctcgtgggca gcgtccgggc caccgccgct gctcctttac ctttcgcttc cttctttcct
     6961 catcgtccag atgcttctcc ttctcggcct cagccttcac gtccggcatt cgtggtgccc
     7021 ggagcccgac gccccgcgtc cggacctgga ggcagccctg ggtctccgga tcaggccagc
     7081 ggccaaaggg tcgccgcacg cacctgttcc cagggcctcc acatcatggc ccctccctcg
     7141 ggttacccca cagcctaggc cgattcgacc tctctccgct ggggccctcg ctggcgtccc
     7201 tgcaccctgg gagcgcgagc ggcgcgcggg cggggaagcg cggcccagac ccccgggtcc
     7261 gcccggagca gctgcgctgt cggggccagg ccgggctccc agtggattcg cgggcacaga
     7321 cgcccaggac cgcgcttccc acgtggcgga gggactgggg acccgggcac ccgtcctgcc
     7381 ccttcacctt ccagctccgc ctcctccgcg cggaccccgc cccgtcccga cccctcccgg
     7441 gtccccggcc cagccccctc cgggccctcc cagcccctcc ccttcctttc cgcggccccg
     7501 ccctctcctc gcggcgcgag tttcaggcag cgctgcgtcc tgctgcgcac gtgggaagcc
     7561 ctggccccgg ccacccccgc gatgccgcgc gctccccgct gccgagc
//
