LOCUS       sequence-169529-        7456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly luciferase under the control of IRF7 5'UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7456)
  AUTHORS   Gandin V, Masvidal L, Hulea L, Gravel SP, Cargnello M, McLaughlan S,
            Cai Y, Balanathan P, Morita M, Rajakumar A, Furic L, Pollak M, Porco
            JA Jr, St-Pierre J, Pelletier J, Larsson O, Topisirovic I
  TITLE     nanoCAGE reveals 5' UTR features that define specific modes of 
            translation of functionally related MTOR-sensitive mRNAs.
  JOURNAL   Genome Res. 2016 May;26(5):636-48. doi: 10.1101/gr.197566.115. Epub 
            2016 Mar 16.
  PUBMED    26984228
REFERENCE   2  (bases 1 to 7456)
  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"; doi: 
            "10.1101/gr.197566.115"; journalName: "Genome Res"; date: "2016-05";
            volume: "26"; issue: "5"; pages: "636-48"
COMMENT     Sequence Label: pCDNA3 IRF7(5'UTR)-FF
FEATURES             Location/Qualifiers
     source          1..7456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        35..414
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        415..618
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     569..589
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     663..682
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        663..681
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1074..2726
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(1140..1158)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     promoter        complement(2809..2827)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2810..2827)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2847..2864)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2853..3077
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3123..3551
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3210..3229)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3420..3441
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3560..3580)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3565..3894
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3745..3880
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3807..3826
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3961..4755
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4015..4034)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4625..4644
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4929..5050
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4966..4985)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5020..5039
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5099..5115)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5099..5115)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5112..5134)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5123..5139
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5147..5153,5154..5171,5172..5177))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5192..5213
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5330..5347)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5501..6089)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5581..5600)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6260..7051,7052..7120))
                     /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     6883..6902
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7121..7225)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7300..7319)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
       61 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      121 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      181 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      241 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      301 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      361 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      421 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      481 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      541 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      601 gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
      661 attaatacga ctcactatag ggagacccaa gcttggtacc gagctcggat ccatcacgtg
      721 tgcgctcccg ctacatctgt agtcacatct acagccacct tggcagagga atctacactg
      781 agttctgaac ctaggtgacc acaagttctc aaacctcatc tgtgaccctc aacaccctaa
      841 tacctgggtc acacttgtga aactgaaatt cctacctgtt accaacactt gtaacgttta
      901 aaacacattt gtgaaacttg gtcacactat ctgtggctac aacctaaaac catagaggca
      961 cccaaggggc cttatttgca atagctgact gctccagtga ctacaaggca tcacagagta
     1021 gtagcatcta ctttgattct cccagactgc ctgtgtagac ggagcggatc caaatggaag
     1081 acgccaaaaa cataaagaaa ggcccggcgc cattctatcc tctagaggat ggaaccgctg
     1141 gagagcaact gcataaggct atgaagagat acgccctggt tcctggaaca attgctttta
     1201 cagatgcaca tatcgaggtg aacatcacgt acgcggaata cttcgaaatg tccgttcggt
     1261 tggcagaagc tatgaaacga tatgggctga atacaaatca cagaatcgtc gtatgcagtg
     1321 aaaactctct tcaattcttt atgccggtgt tgggcgcgtt atttatcgga gttgcagttg
     1381 cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa cagtatgaac atttcgcagc
     1441 ctaccgtagt gtttgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaaat
     1501 taccaataat ccagaaaatt attatcatgg attctaaaac ggattaccag ggatttcagt
     1561 cgatgtacac gttcgtcaca tctcatctac ctcccggttt taatgaatac gattttgtac
     1621 cagagtcctt tgatcgtgac aaaacaattg cactgataat gaattcctct ggatctactg
     1681 ggttacctaa gggtgtggcc cttccgcata gaactgcctg cgtcagattc tcgcatgcca
     1741 gagatcctat ttttggcaat caaatcattc cggatactgc gattttaagt gttgttccat
     1801 tccatcacgg ttttggaatg tttactacac tcggatattt gatatgtgga tttcgagtcg
     1861 tcttaatgta tagatttgaa gaagagctgt ttttacgatc ccttcaggat tacaaaattc
     1921 aaagtgcgtt gctagtacca accctatttt cattcttcgc caaaagcact ctgattgaca
     1981 aatacgattt atctaattta cacgaaattg cttctggggg cgcacctctt tcgaaagaag
     2041 tcggggaagc ggttgcaaaa cgcttccatc ttccagggat acgacaagga tatgggctca
     2101 ctgagactac atcagctatt ctgattacac ccgaggggga tgataaaccg ggcgcggtcg
     2161 gtaaagttgt tccatttttt gaagcgaagg ttgtggatct ggataccggg aaaacgctgg
     2221 gcgttaatca gagaggcgaa ttatgtgtca gaggacctat gattatgtcc ggttatgtaa
     2281 acaatccgga agcgaccaac gccttgattg acaaggatgg atggctacat tctggagaca
     2341 tagcttactg ggacgaagac gaacacttct tcatagttga ccgcttgaag tctttaatta
     2401 aatacaaagg atatcaggtg gcccccgctg aattggaatc gatattgtta caacacccca
     2461 acatcttcga cgcgggcgtg gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg
     2521 ccgttgttgt tttggagcac ggaaagacga tgacggaaaa agagatcgtg gattacgtcg
     2581 ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac
     2641 cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca
     2701 agaagggcgg aaagtccaaa ttgtaaaatg taactgtatt cagcgatgac gaaattctta
     2761 gctattgtaa tcctccgagg cctcgagcat gcatctagag ggccctattc tatagtgtca
     2821 cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2881 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2941 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3001 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     3061 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     3121 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3181 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3241 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3301 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3361 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3421 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3481 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3541 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3601 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3661 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3721 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3781 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     3841 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     3901 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     3961 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4021 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4081 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4141 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4201 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4261 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4321 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4381 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4441 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4501 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4561 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4621 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4681 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4741 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4801 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4861 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4921 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4981 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5041 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5101 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5161 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5221 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5281 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5341 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5401 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5461 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5521 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5581 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5641 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5701 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5761 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5821 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5881 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5941 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6001 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6061 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6121 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6181 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6241 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6301 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6361 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6421 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6481 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6541 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6601 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6661 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6721 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6781 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6841 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6901 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6961 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7021 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7081 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7141 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7201 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     7261 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc
     7321 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     7381 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     7441 ggttaggcgt tttgcg
//
