LOCUS       sequence-337238-        7236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty vector for Firefly-Renilla luciferase assay for hTERT promoter
            activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7236)
  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 7236)
  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
FEATURES             Location/Qualifiers
     source          1..7236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             34..1686
                     /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(100..118)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1727..1848
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1738..1757)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1792..1811
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2149..2169
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(2321..2338)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2492..3080)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2572..2591)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3251..4042,4043..4111))
                     /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     3874..3893
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4112..4216)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4243..4698
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4330..4349)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4540..4561
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4829..4877
                     /note="synthetic polyadenylation signal"
     misc_feature    4891..4982
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4931..4950
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        5037..5233
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5084..5219
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          5265..5397
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     5380..5399
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        5442..5460
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5508..6443
                     /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(6457..6473)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6458..6474)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 agccactacc gcgaggtgct gccgctgtcc cccatggaag acgccaaaaa cataaagaaa
       61 ggcccggcgc cattctatcc gctggaagat ggaaccgctg gagagcaact gcataaggct
      121 atgaagagat acgccctggt tcctggaaca attgctttta cagatgcaca tatcgaggtg
      181 gacatcactt acgctgagta cttcgaaatg tccgttcggt tggcagaagc tatgaaacga
      241 tatgggctga atacaaatca cagaatcgtc gtatgcagtg aaaactctct tcaattcttt
      301 atgccggtgt tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa cgacatttat
      361 aatgaacgtg aattgctcaa cagtatgggc atttcgcagc ctaccgtggt gttcgtttcc
      421 aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaagc tcccaatcat ccaaaaaatt
      481 attatcatgg attctaaaac ggattaccag ggatttcagt cgatgtacac gttcgtcaca
      541 tctcatctac ctcccggttt taatgaatac gattttgtgc cagagtcctt cgatagggac
      601 aagacaattg cactgatcat gaactcctct ggatctactg gtctgcctaa aggtgtcgct
      661 ctgcctcata gaactgcctg cgtgagattc tcgcatgcca gagatcctat ttttggcaat
      721 caaatcattc cggatactgc gattttaagt gttgttccat tccatcacgg ttttggaatg
      781 tttactacac tcggatattt gatatgtgga tttcgagtcg tcttaatgta tagatttgaa
      841 gaagagctgt ttctgaggag ccttcaggat tacaagattc aaagtgcgct gctggtgcca
      901 accctattct ccttcttcgc caaaagcact ctgattgaca aatacgattt atctaattta
      961 cacgaaattg cttctggtgg cgctcccctc tctaaggaag tcggggaagc ggttgccaag
     1021 aggttccatc tgccaggtat caggcaagga tatgggctca ctgagactac atcagctatt
     1081 ctgattacac ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt
     1141 gaagcgaagg ttgtggatct ggataccggg aaaacgctgg gcgttaatca aagaggcgaa
     1201 ctgtgtgtga gaggtcctat gattatgtcc ggttatgtaa acaatccgga agcgaccaac
     1261 gccttgattg acaaggatgg atggctacat tctggagaca tagcttactg ggacgaagac
     1321 gaacacttct tcatcgttga ccgcctgaag tctctgatta agtacaaagg ctatcaggtg
     1381 gctcccgctg aattggaatc catcttgctc caacacccca acatcttcga cgcaggtgtc
     1441 gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg ccgttgttgt tttggagcac
     1501 ggaaagacga tgacggaaaa agagatcgtg gattacgtcg ccagtcaagt aacaaccgcg
     1561 aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct taccggaaaa
     1621 ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca agaagggcgg aaagatcgcc
     1681 gtgtaattct agagtcgggg cggccggccg cttcgagcag acatgataag atacattgat
     1741 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     1801 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     1861 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     1921 aacctctaca aatgtggtaa aatcgataag gatctgaacg atggagcgga gaatgggcgg
     1981 aactgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca
     2041 tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac taattgagat
     2101 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac
     2161 acacattcca cagcggatcc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc
     2221 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
     2281 gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc
     2341 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2401 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2461 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2521 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2581 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2641 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2701 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2761 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2821 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2881 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2941 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3001 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3061 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3121 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3181 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3241 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3301 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3361 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3421 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3481 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3541 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3601 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3661 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3721 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3781 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3841 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3901 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3961 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4021 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4081 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4141 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4201 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg
     4261 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4321 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4381 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4441 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4501 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4561 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4621 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4681 tattaacgct tacaatttgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     4741 cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc atgataagta agtaatatta
     4801 aggtacggga ggtacttgga gcggccgcaa taaaatatct ttattttcat tacatctgtg
     4861 tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa
     4921 acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct
     4981 ctatcgatag gtaccgagct cttacgcgtg ctagcccggg ctcgagatct gcgatctgca
     5041 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     5101 gcccagttcc gcccattctc cgccccatcg ctgactaatt ttttttattt atgcagaggc
     5161 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     5221 aggcttttgc aaaaagctga agttggtcgt gaggcactgg gcaggtaagt atcaaggtta
     5281 caagacaggt ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc
     5341 gtttctgata ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg
     5401 tccactccca gttcaattac agctcttaag gctagagtac ttaatacgac tcactatagg
     5461 ctaggcttgg cattccggta ctgaattgat atcaagctct agccaccatg acttcgaaag
     5521 tttatgatcc agaacaaagg aaacggatga taactggtcc gcagtggtgg gccagatgta
     5581 aacaaatgaa tgttcttgat tcatttatta attattatga ttcagaaaaa catgcagaaa
     5641 atgctgttat ttttttacat ggtaacgcgg cctcttctta tttatggcga catgttgtgc
     5701 cacatattga gccagtagcg cggtgtatta taccagacct tattggtatg ggcaaatcag
     5761 gcaaatctgg taatggttct tataggttac ttgatcatta caaatatctt actgcatggt
     5821 ttgaacttct taatttacca aagaagatca tttttgtcgg ccatgattgg ggtgcttgtt
     5881 tggcatttca ttatagctat gagcatcaag ataagatcaa agcaatagtt cacgctgaaa
     5941 gtgtagtaga tgtgattgaa tcatgggatg aatggcctga tattgaagaa gatattgcgt
     6001 tgatcaaatc tgaagaagga gaaaaaatgg ttttggagaa taacttcttc gtggaaacca
     6061 tgttgccatc aaaaatcatg agaaagttag aaccagaaga atttgcagca tatcttgaac
     6121 cattcaaaga gaaaggtgaa gttcgtcgtc caacattatc atggcctcgt gaaatcccgt
     6181 tagtaaaagg tggtaaacct gacgttgtac aaattgttag gaattataat gcttatctac
     6241 gtgcaagtga tgatttacca aaaatgttta ttgaatcgga cccaggattc ttttccaatg
     6301 ctattgttga aggtgccaag aagtttccta atactgaatt tgtcaaagta aaaggtcttc
     6361 atttttcgca agaagatgca cctgatgaaa tgggaaaata tatcaaatcg ttcgttgagc
     6421 gagttctcaa aaatgaacaa taattctaga gcttatcgat accgtcgacc tcgaatcact
     6481 agtcagctgg aattcagatc tggtaccgag ctccccgggc tgcaggatat cctgcagccc
     6541 ggggacctcg gtaccagatc taggatccag gatccggcga cactccacca tggaagtcct
     6601 cagccttcac gtccggcatt cgtggtgccc ggagcccgac gccccgcgtc cggacctgga
     6661 ggcagccctg ggtctccgga tcaggccagc ggccaaaggg tcgccgcacg cacctgttcc
     6721 cagggcctcc acatcatggc ccctccctcg ggttacccca cagcctaggc cgattcgacc
     6781 tctctccgct ggggccctcg ctggcgtccc tgcaccctgg gagcgcgagc ggcgcgcggg
     6841 cggggaagcg cggcccagac ccccgggtcc gcccggagca gctgcgctgt cggggccagg
     6901 ccgggctccc agtggattcg cgggcacaga cgcccaggac cgcgcttccc acgtggcgga
     6961 gggactgggg acccgggcac ccgtcctgcc ccttcacctt ccagctccgc ctcctccgcg
     7021 cggaccccgc cccgtcccga cccctcccgg gtccccggcc cagccccctc cgggccctcc
     7081 cagcccctcc ccttcctttc cgcggccccg ccctctcctc gcggcgcgag tttcaggcag
     7141 cgctgcgtcc tgctgcgcac gtgggaagcc ctggccccgg ccacccccgc gatgccgcgc
     7201 gctccccgct gccgagccgt gcgctccctg ctgcgc
//
