LOCUS       sequence-448445-        6743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control retroviral vector expressing firefly luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6743)
  AUTHORS   Toyama EQ, Herzig S, Courchet J, Lewis TL Jr, Loson OC, Hellberg K, 
            Young NP, Chen H, Polleux F, Chan DC, Shaw RJ
  TITLE     Metabolism. AMP-activated protein kinase mediates mitochondrial 
            fission in response to energy stress.
  JOURNAL   Science. 2016 Jan 15;351(6270):275-81. doi: 10.1126/science.aab4138.
  PUBMED    26816379
REFERENCE   2  (bases 1 to 6743)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2016-01-15- 15"; volume: "351"; 
            issue: "6270"; pages: "275-81"
COMMENT     Sequence Label: pQCXIB Firefly_Luciferase
FEATURES             Location/Qualifiers
     source          1..6743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    176..194
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    197..215
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    256..280
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             292..333
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      403..412
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             409..2061
                     /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(475..493)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     protein_bind    complement(2068..2167)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2184..2203)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2185..2203)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2208..2224)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2208..2224)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2326..2345)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2337..3146
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2404..2423)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(join(3580..4371,4372..4440))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4203..4222
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4441..4545)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4613..4631
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        4731..5110
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5111..5314
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             5315..5490
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    5553..5752
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5953..6369
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6299..6321
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6337..6353
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        join(6714..6743,1..174)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
       61 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      121 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct
      181 atcagtgata gagatctccc tatcagtgat agagatcgtc gactagtcca gtgtggtgga
      241 attctgcaga tatcaacaag tttgtacaaa aaagcaggct ccaccatgga aggtaagcct
      301 atccctaacc ctctcctcgg tctcgattct acgggatcca tgggaaccaa ttcagtccct
      361 aggcttttgc aaaaagcttg gcattccggt actgttggta aagccaccat ggaagacgcc
      421 aaaaacataa agaaaggccc ggcgccattc tatccgctgg aagatggaac cgctggagag
      481 caactgcata aggctatgaa gagatacgcc ctggttcctg gaacaattgc ttttacagat
      541 gcacatatcg aggtggacat cacttacgct gagtacttcg aaatgtccgt tcggttggca
      601 gaagctatga aacgatatgg gctgaataca aatcacagaa tcgtcgtatg cagtgaaaac
      661 tctcttcaat tctttatgcc ggtgttgggc gcgttattta tcggagttgc agttgcgccc
      721 gcgaacgaca tttataatga acgtgaattg ctcaacagta tgggcatttc gcagcctacc
      781 gtggtgttcg tttccaaaaa ggggttgcaa aaaattttga acgtgcaaaa aaagctccca
      841 atcatccaaa aaattattat catggattct aaaacggatt accagggatt tcagtcgatg
      901 tacacgttcg tcacatctca tctacctccc ggttttaatg aatacgattt tgtgccagag
      961 tccttcgata gggacaagac aattgcactg atcatgaact cctctggatc tactggtctg
     1021 cctaaaggtg tcgctctgcc tcatagaact gcctgcgtga gattctcgca tgccagagat
     1081 cctatttttg gcaatcaaat cattccggat actgcgattt taagtgttgt tccattccat
     1141 cacggttttg gaatgtttac tacactcgga tatttgatat gtggatttcg agtcgtctta
     1201 atgtatagat ttgaagaaga gctgtttctg aggagccttc aggattacaa gattcaaagt
     1261 gcgctgctgg tgccaaccct attctccttc ttcgccaaaa gcactctgat tgacaaatac
     1321 gatttatcta atttacacga aattgcttct ggtggcgctc ccctctctaa ggaagtcggg
     1381 gaagcggttg ccaagaggtt ccatctgcca ggtatcaggc aaggatatgg gctcactgag
     1441 actacatcag ctattctgat tacacccgag ggggatgata aaccgggcgc ggtcggtaaa
     1501 gttgttccat tttttgaagc gaaggttgtg gatctggata ccgggaaaac gctgggcgtt
     1561 aatcaaagag gcgaactgtg tgtgagaggt cctatgatta tgtccggtta tgtaaacaat
     1621 ccggaagcga ccaacgcctt gattgacaag gatggatggc tacattctgg agacatagct
     1681 tactgggacg aagacgaaca cttcttcatc gttgaccgcc tgaagtctct gattaagtac
     1741 aaaggctatc aggtggctcc cgctgaattg gaatccatct tgctccaaca ccccaacatc
     1801 ttcgacgcag gtgtcgcagg tcttcccgac gatgacgccg gtgaacttcc cgccgccgtt
     1861 gttgttttgg agcacggaaa gacgatgacg gaaaaagaga tcgtggatta cgtcgccagt
     1921 caagtaacaa ccgcgaaaaa gttgcgcgga ggagttgtgt ttgtggacga agtaccgaaa
     1981 ggtcttaccg gaaaactcga cgcaagaaaa atcagagaga tcctcataaa ggccaagaag
     2041 ggcggaaaga tcgccgtgta attctagacc cagctttctt gtacaaagtt ggcattataa
     2101 gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca
     2161 ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt
     2221 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     2281 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     2341 gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat
     2401 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt
     2461 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     2521 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     2581 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggaa
     2641 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     2701 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     2761 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     2821 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     2881 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     2941 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     3001 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     3061 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     3121 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga
     3181 gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt
     3241 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3301 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3361 tttgtttgcc ggcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3421 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3481 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3541 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3601 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3661 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3721 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3781 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3841 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg
     3901 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3961 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4021 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4081 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4141 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg
     4201 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4261 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4321 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4381 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4441 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4501 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4561 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     4621 tatcacgagg ccctttcgtc ttcaagaatt agcttggcca ttgcatacgt tgtatccata
     4681 tcataatatg tacatttata ttggctcatg tccaacatta ccgccatgtt gacattgatt
     4741 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     4801 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     4861 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     4921 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     4981 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     5041 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     5101 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     5161 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     5221 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     5281 gcgtgtacgg tgggaggtct atataagcag agctcaataa aagagcccac aacccctcac
     5341 tcggcgcgcc agtcttccga tagactgcgt cgcccgggta cccgtattcc caataaagcc
     5401 tcttgctgtt tgcatccgaa tcgtggtctc gctgttcctt gggagggtct cctctgagtg
     5461 attgactacc cacgacgggg gtctttcatt tgggggctcg tccgggattt ggagacccct
     5521 gcccagggac caccgaccca ccaccgggag gtaagctggc cagcaactta tctgtgtctg
     5581 tccgattgtc tagtgtctat gtttgatgtt atgcgcctgc gtctgtacta gttagctaac
     5641 tagctctgta tctggcggac ccgtggtgga actgacgagt tctgaacacc cggccgcaac
     5701 cctgggagac gtcccaggga ctttgggggc cgtttttgtg gcccgacctg aggaagggag
     5761 tcgatgtgga atccgacccc gtcaggatat gtggttctgg taggagacga gaacctaaaa
     5821 cagttcccgc ctccgtctga atttttgctt tcggtttgga accgaagccg cgcgtcttgt
     5881 ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg
     5941 tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg tcactggaaa
     6001 gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc
     6061 ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac
     6121 cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca
     6181 gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc
     6241 aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc gtctctcccc
     6301 cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct cactccttct
     6361 ctaggcgccg gaattgaaga tctgggggat cgataagctt gggagttccg cttacataac
     6421 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     6481 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     6541 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     6601 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     6661 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     6721 ggttttggca gtacatcaat ggg
//
