LOCUS       sequence-207856-        6658 bp    DNA     circular SYN 15-APR-2025
DEFINITION  luciferase reporter, CMV promoter (3 NFkB sites mutated).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6658)
  AUTHORS   He B, Weber GF
  TITLE     Synergistic activation of the CMV promoter by NF-kappaB P50 and PKG.
  JOURNAL   Biochem Biophys Res Commun. 2004 Aug 13;321(1):13-20. doi: 
            10.1016/j.bbrc.2004.06.101.
  PUBMED    15358208
REFERENCE   2  (bases 1 to 6658)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.bbrc.2004.06"; journalName: "Biochem Biophys Res Commun";
            date: "2004-08-13- 13"; volume: "321"; issue: "1"; pages: "13-20"
COMMENT     Sequence Label: pCR3.1-mutLuc
FEATURES             Location/Qualifiers
     source          1..6658
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..583
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     534..554
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     628..647
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        628..646
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      705..714
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             711..2363
                     /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(777..795)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(2396..2413)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2402..2626
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2758..3346)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2838..2857)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3675..3722
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3747..3766
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3954..4748)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(3954..4748)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4065..4084)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4675..4694
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4783..5140)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4797..4932
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4949..5140
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5097..5117
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(5200..5991,5992..6060))
                     /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     5823..5842
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6061..6165)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6192..6647
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6279..6298)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6489..6510
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaatactca
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatctc actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacctcac tttcaaaatg tcgtaacaac tccgccccat tgacgcaaat
      541 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga
      601 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg
      661 ctagcgttta aacttaagct tggcattccg gtactgttgg taaagccacc atggaagacg
      721 ccaaaaacat aaagaaaggc ccggcgccat tctatccgct ggaagatgga accgctggag
      781 agcaactgca taaggctatg aagagatacg ccctggttcc tggaacaatt gcttttacag
      841 atgcacatat cgaggtggac atcacttacg ctgagtactt cgaaatgtcc gttcggttgg
      901 cagaagctat gaaacgatat gggctgaata caaatcacag aatcgtcgta tgcagtgaaa
      961 actctcttca attctttatg ccggtgttgg gcgcgttatt tatcggagtt gcagttgcgc
     1021 ccgcgaacga catttataat gaacgtgaat tgctcaacag tatgggcatt tcgcagccta
     1081 ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa aaaaagctcc
     1141 caatcatcca aaaaattatt atcatggatt ctaaaacgga ttaccaggga tttcagtcga
     1201 tgtacacgtt cgtcacatct catctacctc ccggttttaa tgaatacgat tttgtgccag
     1261 agtccttcga tagggacaag acaattgcac tgatcatgaa ctcctctgga tctactggtc
     1321 tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt gagattctcg catgccagag
     1381 atcctatttt tggcaatcaa atcattccgg atactgcgat tttaagtgtt gttccattcc
     1441 atcacggttt tggaatgttt actacactcg gatatttgat atgtggattt cgagtcgtct
     1501 taatgtatag atttgaagaa gagctgtttc tgaggagcct tcaggattac aagattcaaa
     1561 gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa aagcactctg attgacaaat
     1621 acgatttatc taatttacac gaaattgctt ctggtggcgc tcccctctct aaggaagtcg
     1681 gggaagcggt tgccaagagg ttccatctgc caggtatcag gcaaggatat gggctcactg
     1741 agactacatc agctattctg attacacccg agggggatga taaaccgggc gcggtcggta
     1801 aagttgttcc attttttgaa gcgaaggttg tggatctgga taccgggaaa acgctgggcg
     1861 ttaatcaaag aggcgaactg tgtgtgagag gtcctatgat tatgtccggt tatgtaaaca
     1921 atccggaagc gaccaacgcc ttgattgaca aggatggatg gctacattct ggagacatag
     1981 cttactggga cgaagacgaa cacttcttca tcgttgaccg cctgaagtct ctgattaagt
     2041 acaaaggcta tcaggtggct cccgctgaat tggaatccat cttgctccaa caccccaaca
     2101 tcttcgacgc aggtgtcgca ggtcttcccg acgatgacgc cggtgaactt cccgccgccg
     2161 ttgttgtttt ggagcacgga aagacgatga cggaaaaaga gatcgtggat tacgtcgcca
     2221 gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt gtttgtggac gaagtaccga
     2281 aaggtcttac cggaaaactc gacgcaagaa aaatcagaga gatcctcata aaggccaaga
     2341 agggcggaaa gatcgccgtg taattctaga gggcccgttt aaacccgctg atcagcctcg
     2401 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2461 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2521 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2581 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     2641 agaaccagtg gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2701 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2761 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2821 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2881 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2941 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3001 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3061 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3121 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3181 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3241 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3301 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3361 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3421 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3481 aatctaaagt atatatgagt aacctgaggc tatggcaggg cctgccgccc cgacgttggc
     3541 tgcgagccct gggccttcac ccgaacttgg ggggtggggt ggggaaaagg aagaaacgcg
     3601 ggcgtattgg ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg
     3661 aaccccgcgt ttatgaacaa acgacccaac accgtgcgtt ttattctgtc tttttattgc
     3721 cgtcatagcg cgggttcctt ccggtattgt ctccttccgt gtttcagtta gcctccccct
     3781 agggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc
     3841 ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg
     3901 tgatggcagg ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag
     3961 aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa
     4021 agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc
     4081 aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa
     4141 aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga
     4201 tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc
     4261 tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct
     4321 cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc
     4381 agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac
     4441 aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca
     4501 acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc
     4561 tcgtcttgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc
     4621 ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag
     4681 tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt
     4741 tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa agcctaggcc
     4801 tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc
     4861 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
     4921 taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat
     4981 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta
     5041 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc
     5101 taactgacac acattccaca gctggttctt tccgcctcag gactcttcct ttttcaataa
     5161 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5221 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5281 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5341 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5401 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5461 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5521 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5581 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5641 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5701 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5761 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5821 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5881 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5941 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6001 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6061 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6121 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6181 agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     6241 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     6301 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     6361 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     6421 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     6481 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     6541 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     6601 acaaaaattt aacgcgaatt ttaacaaaat attagcgctt acaatttacg cgcgcgtt
//
