LOCUS       sequence-337252-        7379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly-Renilla luciferase assay for hTERT promoter activity of 
            hTERT intron-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7379)
  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 7379)
  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 hTERT intron-1
FEATURES             Location/Qualifiers
     source          1..7379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(739..755)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(740..756)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             1695..3347
                     /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(1761..1779)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3388..3509
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3399..3418)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3453..3472
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3810..3830
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(3982..3999)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4153..4741)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4233..4252)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4912..5703,5704..5772))
                     /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     5535..5554
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5773..5877)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5904..6359
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5991..6010)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6201..6222
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6490..6538
                     /note="synthetic polyadenylation signal"
     misc_feature    6552..6643
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6592..6611
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        6698..6894
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6745..6880
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          6926..7058
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     7041..7060
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        7103..7121
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(7169..7379,1..725)
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
ORIGIN
        1 cgcggtgtat tataccagac cttattggta tgggcaaatc aggcaaatct ggtaatggtt
       61 cttataggtt acttgatcat tacaaatatc ttactgcatg gtttgaactt cttaatttac
      121 caaagaagat catttttgtc ggccatgatt ggggtgcttg tttggcattt cattatagct
      181 atgagcatca agataagatc aaagcaatag ttcacgctga aagtgtagta gatgtgattg
      241 aatcatggga tgaatggcct gatattgaag aagatattgc gttgatcaaa tctgaagaag
      301 gagaaaaaat ggttttggag aataacttct tcgtggaaac catgttgcca tcaaaaatca
      361 tgagaaagtt agaaccagaa gaatttgcag catatcttga accattcaaa gagaaaggtg
      421 aagttcgtcg tccaacatta tcatggcctc gtgaaatccc gttagtaaaa ggtggtaaac
      481 ctgacgttgt acaaattgtt aggaattata atgcttatct acgtgcaagt gatgatttac
      541 caaaaatgtt tattgaatcg gacccaggat tcttttccaa tgctattgtt gaaggtgcca
      601 agaagtttcc taatactgaa tttgtcaaag taaaaggtct tcatttttcg caagaagatg
      661 cacctgatga aatgggaaaa tatatcaaat cgttcgttga gcgagttctc aaaaatgaac
      721 aataattcta gagcttatcg ataccgtcga cctcgaatca ctagtcagct ggaattcaga
      781 tctggtaccg agctccccgg gctgcaggat atcctgcagc ccggggacct cggtaccaga
      841 tctaggatcc aggatccggc gacactccac catggaagtc ctcagggggc caccagctcc
      901 ttcaggcagg acacctgcgg gggaagcgcc ctgagtcgcc tgcgctgctc tccgcatgtc
      961 gctggttccc cccggccgcc ctcaacccca gccggacgcc gaccccgggg aggcccacct
     1021 ggcctcagcc ttcacgtccg gcattcgtgg tgcccggagc ccgacgcccc gcgtccggac
     1081 ctggaggcag ccctgggtct ccggatcagg ccagcggcca aagggtcgcc gcacgcacct
     1141 gttcccaggg cctccacatc atggcccctc cctcgggtta ccccacagcc taggccgatt
     1201 cgacctctct ccgctggggc cctcgctggc gtccctgcac cctgggagcg cgagcggcgc
     1261 gcgggcgggg aagcgcggcc cagacccccg ggtccgcccg gagcagctgc gctgtcgggg
     1321 ccaggccggg ctcccagtgg attcgcgggc acagacgccc aggaccgcgc ttcccacgtg
     1381 gcggagggac tggggacccg ggcacccgtc ctgccccttc accttccagc tccgcctcct
     1441 ccgcgcggac cccgccccgt cccgacccct cccgggtccc cggcccagcc ccctccgggc
     1501 cctcccagcc cctccccttc ctttccgcgg ccccgccctc tcctcgcggc gcgagtttca
     1561 ggcagcgctg cgtcctgctg cgcacgtggg aagccctggc cccggccacc cccgcgatgc
     1621 cgcgcgctcc ccgctgccga gccgtgcgct ccctgctgcg cagccactac cgcgaggtgc
     1681 tgccgctgtc ccccatggaa gacgccaaaa acataaagaa aggcccggcg ccattctatc
     1741 cgctggaaga tggaaccgct ggagagcaac tgcataaggc tatgaagaga tacgccctgg
     1801 ttcctggaac aattgctttt acagatgcac atatcgaggt ggacatcact tacgctgagt
     1861 acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg atatgggctg aatacaaatc
     1921 acagaatcgt cgtatgcagt gaaaactctc ttcaattctt tatgccggtg ttgggcgcgt
     1981 tatttatcgg agttgcagtt gcgcccgcga acgacattta taatgaacgt gaattgctca
     2041 acagtatggg catttcgcag cctaccgtgg tgttcgtttc caaaaagggg ttgcaaaaaa
     2101 ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat tattatcatg gattctaaaa
     2161 cggattacca gggatttcag tcgatgtaca cgttcgtcac atctcatcta cctcccggtt
     2221 ttaatgaata cgattttgtg ccagagtcct tcgataggga caagacaatt gcactgatca
     2281 tgaactcctc tggatctact ggtctgccta aaggtgtcgc tctgcctcat agaactgcct
     2341 gcgtgagatt ctcgcatgcc agagatccta tttttggcaa tcaaatcatt ccggatactg
     2401 cgattttaag tgttgttcca ttccatcacg gttttggaat gtttactaca ctcggatatt
     2461 tgatatgtgg atttcgagtc gtcttaatgt atagatttga agaagagctg tttctgagga
     2521 gccttcagga ttacaagatt caaagtgcgc tgctggtgcc aaccctattc tccttcttcg
     2581 ccaaaagcac tctgattgac aaatacgatt tatctaattt acacgaaatt gcttctggtg
     2641 gcgctcccct ctctaaggaa gtcggggaag cggttgccaa gaggttccat ctgccaggta
     2701 tcaggcaagg atatgggctc actgagacta catcagctat tctgattaca cccgaggggg
     2761 atgataaacc gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag gttgtggatc
     2821 tggataccgg gaaaacgctg ggcgttaatc aaagaggcga actgtgtgtg agaggtccta
     2881 tgattatgtc cggttatgta aacaatccgg aagcgaccaa cgccttgatt gacaaggatg
     2941 gatggctaca ttctggagac atagcttact gggacgaaga cgaacacttc ttcatcgttg
     3001 accgcctgaa gtctctgatt aagtacaaag gctatcaggt ggctcccgct gaattggaat
     3061 ccatcttgct ccaacacccc aacatcttcg acgcaggtgt cgcaggtctt cccgacgatg
     3121 acgccggtga acttcccgcc gccgttgttg ttttggagca cggaaagacg atgacggaaa
     3181 aagagatcgt ggattacgtc gccagtcaag taacaaccgc gaaaaagttg cgcggaggag
     3241 ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca agaaaaatca
     3301 gagagatcct cataaaggcc aagaagggcg gaaagatcgc cgtgtaattc tagagtcggg
     3361 gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     3421 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3481 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3541 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     3601 aaatcgataa ggatctgaac gatggagcgg agaatgggcg gaactgggcg gagttagggg
     3661 cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     3721 gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct
     3781 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagcggatc
     3841 cgtcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
     3901 atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg
     3961 ccggcagcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4021 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4081 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4141 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4201 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4261 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4321 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4381 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4441 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4501 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4561 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4621 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4681 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4741 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4801 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4861 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4921 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4981 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5041 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5101 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5161 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5221 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5281 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5341 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5401 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5461 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5521 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5581 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5641 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5701 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5761 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5821 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5881 atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5941 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     6001 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     6061 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     6121 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     6181 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     6241 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     6301 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg
     6361 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     6421 attacgccag cccaagctac catgataagt aagtaatatt aaggtacggg aggtacttgg
     6481 agcggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa
     6541 tcgatagtac taacatacgc tctccatcaa aacaaaacga aacaaaacaa actagcaaaa
     6601 taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc tctatcgata ggtaccgagc
     6661 tcttacgcgt gctagcccgg gctcgagatc tgcgatctgc atctcaatta gtcagcaacc
     6721 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     6781 ccgccccatc gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     6841 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctg
     6901 aagttggtcg tgaggcactg ggcaggtaag tatcaaggtt acaagacagg tttaaggaga
     6961 ccaatagaaa ctgggcttgt cgagacagag aagactcttg cgtttctgat aggcacctat
     7021 tggtcttact gacatccact ttgcctttct ctccacaggt gtccactccc agttcaatta
     7081 cagctcttaa ggctagagta cttaatacga ctcactatag gctaggcttg gcattccggt
     7141 actgaattga tatcaagctc tagccaccat gacttcgaaa gtttatgatc cagaacaaag
     7201 gaaacggatg ataactggtc cgcagtggtg ggccagatgt aaacaaatga atgttcttga
     7261 ttcatttatt aattattatg attcagaaaa acatgcagaa aatgctgtta tttttttaca
     7321 tggtaacgcg gcctcttctt atttatggcg acatgttgtg ccacatattg agccagtag
//
