LOCUS       sequence-169530-        7105 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly luciferase under the control of ATP5O TISU.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7105)
  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 7105)
  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 ATP5O(TISU)-FF
FEATURES             Location/Qualifiers
     source          1..7105
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        36..415
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        416..619
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     570..590
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     664..683
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        664..682
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             724..2376
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MAAAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(790..808)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     promoter        complement(2459..2477)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2460..2477)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2497..2514)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2503..2727
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2773..3201
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2860..2879)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3070..3091
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3210..3230)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3215..3544
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3395..3530
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3457..3476
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3611..4405
                     /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(3665..3684)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4275..4294
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4579..4700
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4616..4635)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4670..4689
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4749..4765)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4749..4765)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4762..4784)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4773..4789
                     /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(4797..4803,4804..4821,4822..4827))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4842..4863
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4980..4997)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5151..5739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5231..5250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5910..6701,6702..6770))
                     /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     6533..6552
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6771..6875)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6950..6969)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
       61 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      121 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      181 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      241 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      301 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      361 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      421 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      481 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      541 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      601 ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga
      661 aattaatacg actcactata gggagaccca agcttggtac cgagctcgga tccaagggag
      721 aagatggctg ccgccaaaaa cataaagaaa ggcccggcgc cattctatcc tctagaggat
      781 ggaaccgctg gagagcaact gcataaggct atgaagagat acgccctggt tcctggaaca
      841 attgctttta cagatgcaca tatcgaggtg aacatcacgt acgcggaata cttcgaaatg
      901 tccgttcggt tggcagaagc tatgaaacga tatgggctga atacaaatca cagaatcgtc
      961 gtatgcagtg aaaactctct tcaattcttt atgccggtgt tgggcgcgtt atttatcgga
     1021 gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa cagtatgaac
     1081 atttcgcagc ctaccgtagt gtttgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg
     1141 caaaaaaaat taccaataat ccagaaaatt attatcatgg attctaaaac ggattaccag
     1201 ggatttcagt cgatgtacac gttcgtcaca tctcatctac ctcccggttt taatgaatac
     1261 gattttgtac cagagtcctt tgatcgtgac aaaacaattg cactgataat gaattcctct
     1321 ggatctactg ggttacctaa gggtgtggcc cttccgcata gaactgcctg cgtcagattc
     1381 tcgcatgcca gagatcctat ttttggcaat caaatcattc cggatactgc gattttaagt
     1441 gttgttccat tccatcacgg ttttggaatg tttactacac tcggatattt gatatgtgga
     1501 tttcgagtcg tcttaatgta tagatttgaa gaagagctgt ttttacgatc ccttcaggat
     1561 tacaaaattc aaagtgcgtt gctagtacca accctatttt cattcttcgc caaaagcact
     1621 ctgattgaca aatacgattt atctaattta cacgaaattg cttctggggg cgcacctctt
     1681 tcgaaagaag tcggggaagc ggttgcaaaa cgcttccatc ttccagggat acgacaagga
     1741 tatgggctca ctgagactac atcagctatt ctgattacac ccgaggggga tgataaaccg
     1801 ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg ttgtggatct ggataccggg
     1861 aaaacgctgg gcgttaatca gagaggcgaa ttatgtgtca gaggacctat gattatgtcc
     1921 ggttatgtaa acaatccgga agcgaccaac gccttgattg acaaggatgg atggctacat
     1981 tctggagaca tagcttactg ggacgaagac gaacacttct tcatagttga ccgcttgaag
     2041 tctttaatta aatacaaagg atatcaggtg gcccccgctg aattggaatc gatattgtta
     2101 caacacccca acatcttcga cgcgggcgtg gcaggtcttc ccgacgatga cgccggtgaa
     2161 cttcccgccg ccgttgttgt tttggagcac ggaaagacga tgacggaaaa agagatcgtg
     2221 gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg
     2281 gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc
     2341 ataaaggcca agaagggcgg aaagtccaaa ttgtaaaatg taactgtatt cagcgatgac
     2401 gaaattctta gctattgtaa tcctccgagg cctcgagcat gcatctagag ggccctattc
     2461 tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc
     2521 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     2581 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     2641 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     2701 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     2761 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2821 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2881 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2941 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     3001 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3061 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     3121 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3181 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3241 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3301 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3361 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3421 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3481 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3541 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     3601 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3661 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3721 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3781 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3841 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     3901 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     3961 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4021 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4081 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     4141 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4201 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4261 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4321 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4381 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4441 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4501 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4561 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4621 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4681 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4741 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4801 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4861 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4921 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4981 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5041 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5101 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5161 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5221 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5281 ctgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5341 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5401 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5461 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5521 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5581 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5641 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5701 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5761 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5821 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5881 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5941 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6001 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6061 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6121 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6181 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6241 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6301 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6361 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6421 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6481 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6541 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6601 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6661 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6721 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6781 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6841 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6901 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     6961 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     7021 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     7081 atctgcttag ggttaggcgt tttgc
//
