LOCUS       sequence-207859-        6655 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  luciferase reporter, CMV promoter (CRE site adjacent to NFkB site 
            mutated).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6655)
  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 6655)
  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: 
            "10.1016/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-CREM2 luc
FEATURES             Location/Qualifiers
     source          1..6655
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..376
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        377..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     531..551
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     625..644
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        625..643
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      702..711
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             708..2360
                     /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(774..792)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(2393..2410)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2399..2623
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2755..3343)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2835..2854)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3672..3719
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3744..3763
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3951..4745)
                     /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(3951..4745)
                     /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(4062..4081)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4672..4691
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4780..5137)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4794..4929
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4946..5137
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5094..5114
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(5197..5988,5989..6057))
                     /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     5820..5839
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6058..6162)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6189..6644
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6276..6295)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6486..6507
                     /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 ccaataggga
      181 ctttccattg caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      241 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      301 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      361 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      421 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      481 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      541 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      601 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
      661 gcgtttaaac ttaagcttgg cattccggta ctgttggtaa agccaccatg gaagacgcca
      721 aaaacataaa gaaaggcccg gcgccattct atccgctgga agatggaacc gctggagagc
      781 aactgcataa ggctatgaag agatacgccc tggttcctgg aacaattgct tttacagatg
      841 cacatatcga ggtggacatc acttacgctg agtacttcga aatgtccgtt cggttggcag
      901 aagctatgaa acgatatggg ctgaatacaa atcacagaat cgtcgtatgc agtgaaaact
      961 ctcttcaatt ctttatgccg gtgttgggcg cgttatttat cggagttgca gttgcgcccg
     1021 cgaacgacat ttataatgaa cgtgaattgc tcaacagtat gggcatttcg cagcctaccg
     1081 tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa aagctcccaa
     1141 tcatccaaaa aattattatc atggattcta aaacggatta ccagggattt cagtcgatgt
     1201 acacgttcgt cacatctcat ctacctcccg gttttaatga atacgatttt gtgccagagt
     1261 ccttcgatag ggacaagaca attgcactga tcatgaactc ctctggatct actggtctgc
     1321 ctaaaggtgt cgctctgcct catagaactg cctgcgtgag attctcgcat gccagagatc
     1381 ctatttttgg caatcaaatc attccggata ctgcgatttt aagtgttgtt ccattccatc
     1441 acggttttgg aatgtttact acactcggat atttgatatg tggatttcga gtcgtcttaa
     1501 tgtatagatt tgaagaagag ctgtttctga ggagccttca ggattacaag attcaaagtg
     1561 cgctgctggt gccaacccta ttctccttct tcgccaaaag cactctgatt gacaaatacg
     1621 atttatctaa tttacacgaa attgcttctg gtggcgctcc cctctctaag gaagtcgggg
     1681 aagcggttgc caagaggttc catctgccag gtatcaggca aggatatggg ctcactgaga
     1741 ctacatcagc tattctgatt acacccgagg gggatgataa accgggcgcg gtcggtaaag
     1801 ttgttccatt ttttgaagcg aaggttgtgg atctggatac cgggaaaacg ctgggcgtta
     1861 atcaaagagg cgaactgtgt gtgagaggtc ctatgattat gtccggttat gtaaacaatc
     1921 cggaagcgac caacgccttg attgacaagg atggatggct acattctgga gacatagctt
     1981 actgggacga agacgaacac ttcttcatcg ttgaccgcct gaagtctctg attaagtaca
     2041 aaggctatca ggtggctccc gctgaattgg aatccatctt gctccaacac cccaacatct
     2101 tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg tgaacttccc gccgccgttg
     2161 ttgttttgga gcacggaaag acgatgacgg aaaaagagat cgtggattac gtcgccagtc
     2221 aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt tgtggacgaa gtaccgaaag
     2281 gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat cctcataaag gccaagaagg
     2341 gcggaaagat cgccgtgtaa ttctagaggg cccgtttaaa cccgctgatc agcctcgact
     2401 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2461 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2521 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2581 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2641 accagtggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2701 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2761 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2821 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2881 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2941 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3001 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3061 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3121 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3181 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3241 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3301 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3361 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3421 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3481 ctaaagtata tatgagtaac ctgaggctat ggcagggcct gccgccccga cgttggctgc
     3541 gagccctggg ccttcacccg aacttggggg gtggggtggg gaaaaggaag aaacgcgggc
     3601 gtattggccc caatggggtc tcggtggggt atcgacagag tgccagccct gggaccgaac
     3661 cccgcgttta tgaacaaacg acccaacacc gtgcgtttta ttctgtcttt ttattgccgt
     3721 catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc tccccctagg
     3781 gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg
     3841 gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtga
     3901 tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac
     3961 tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc
     4021 acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac
     4081 gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag
     4141 cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc
     4201 tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga
     4261 tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc
     4321 tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc
     4381 cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg
     4441 agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg
     4501 tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg
     4561 tcttgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc
     4621 tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca
     4681 tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca
     4741 atcatgcgaa acgatcctca tcctgtctct tgatcgatct ttgcaaaagc ctaggcctcc
     4801 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     4861 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     4921 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
     4981 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
     5041 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     5101 ctgacacaca ttccacagct ggttctttcc gcctcaggac tcttcctttt tcaataaatc
     5161 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5221 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5281 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5341 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5401 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5461 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5521 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5581 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5641 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5701 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5761 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5821 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5881 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5941 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6001 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6061 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6121 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6181 gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     6241 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     6301 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     6361 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     6421 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     6481 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     6541 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     6601 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttacgcgc gcgtt
//
