LOCUS       sequence-209100-        6359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fluorescent fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6359)
  AUTHORS   Schmid JA, Birbach A, Hofer-Warbinek R, Pengg M, Burner U, 
            Furtmuller PG, Binder BR, de Martin R
  TITLE     Dynamics of NF kappa B and Ikappa Balpha studied with green 
            fluorescent protein (GFP) fusion proteins. Investigation of GFP-p65 
            binding to DNa by fluorescence resonance energy transfer.
  JOURNAL   J Biol Chem. 2000 Jun 2;275(22):17035-42. doi: 
            10.1074/jbc.M000291200.
  PUBMED    10747893
REFERENCE   2  (bases 1 to 6359)
  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: "10.1074/jbc"; 
            journalName: "J Biol Chem"; date: "2000-06-2- 2"; volume: "275"; 
            issue: "22"; pages: "17035-42"
COMMENT     Sequence Label: EGFP-p65
FEATURES             Location/Qualifiers
     source          1..6359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2644..3015
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     polyA_signal    3147..3268
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3184..3203)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3238..3257
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3275..3730)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3412..3433)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3624..3643
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3757..3861
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3863..4220
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3863..4054
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3886..3906)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4071..4206
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4255..5049
                     /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             4255..5049
                     /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(4309..4328)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4919..4938
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5237..5256)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5281..5328
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5657..6245
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6146..6165
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcca ccatggacga actgttcccc
     1381 ctcatcttcc cggcagagcc agcccaggcc tctggcccct atgtggagat cattgagcag
     1441 cccaagcagc ggggcatgcg cttccgctac aagtgcgagg ggcgctccgc gggcagcatc
     1501 ccaggcgaga ggagcacaga taccaccaag acccacccca ccatcaagat caatggctac
     1561 acaggaccag ggacagtgcg catctccctg gtcaccaagg accctcctca ccggcctcac
     1621 ccccacgagc ttgtaggaaa ggactgccgg gatggcttct atgaggctga gctctgcccg
     1681 gaccgctgca tccacagttt ccagaacctg ggaatccagt gtgtgaagaa gcgggacctg
     1741 gagcaggcta tcagtcagcg catccagacc aacaacaacc ccttccaagt tcctatagaa
     1801 gagcagcgtg gggactacga cctgaatgct gtgcggctct gcttccaggt gacagtgcgg
     1861 gacccatcag gcaggcccct ccgcctgccg cctgtccttc ctcatcccat ctttgacaat
     1921 cgtgccccca acactgccga gctcaagatc tgccgagtga accgaaactc tggcagctgc
     1981 ctcggtgggg atgagatctt cctactgtgt gacaaggtgc agaaagagga cattgaggtg
     2041 tatttcacgg gaccaggctg ggaggcccga ggctcctttt cgcaagctga tgtgcaccga
     2101 caagtggcca ttgtgttccg gacccctccc tacgcagacc ccagcctgca ggctcctgtg
     2161 cgtgtctcca tgcagctgcg gcggccttcc gaccgggagc tcagtgagcc catggaattc
     2221 cagtacctgc cagatacaga cgatcgtcac cggattgagg agaaacgtaa aaggacatat
     2281 gagaccttca agagcatcat gaagaagagt cctttcagcg gacccaccga cccccggcct
     2341 ccacctcgac gcattgctgt gccttcccgc agctcagctt ctgtccccaa gccagcaccc
     2401 cagccctatc cctttacgtc atccctgagc accatcaact atgatgagtt tcccaccatg
     2461 gtgtttcctt ctgggcagat cagccaggcc tcggccttgg ccccggcccc tccccaagtc
     2521 ctgccccagg ctccagcccc tgcccctgct ccagccatgg tatcagctct ggcccaggcc
     2581 ccagcccctg tcccagtcct agccccaggc cctcctcagg ctgtggcccc acctgccccc
     2641 aagcccaccc aggctgggga aggaacgctg tcagaggccc tgctgcagct gcagtttgat
     2701 gatgaagacc tgggggcctt gcttggcaac agcacagacc cagctgtgtt cacagacctg
     2761 gcatccgtcg acaactccga gtttcagcag ctgctgaacc agggcatacc tgtggccccc
     2821 cacacaactg agcccatgct gatggagtac cctgaggcta taactcgcct agtgacaggg
     2881 gcccagaggc cccccgaccc agctcctgct ccactggggg ccccggggct ccccaatggc
     2941 ctcctttcag gagatgaaga cttctcctcc attgcggaca tggacttctc agccctgctg
     3001 agtcagatca gctccttgga tccaccggat ctagataact gatcataatc agccatacca
     3061 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3121 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3181 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3241 gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta
     3301 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3361 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3421 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3481 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3541 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3601 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3661 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     3721 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3781 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3841 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     3901 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3961 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4021 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4081 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4141 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4201 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4261 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4321 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4381 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     4441 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4501 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4561 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4621 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4681 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4741 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     4801 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4861 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4921 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4981 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5041 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5101 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5161 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5221 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     5281 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     5341 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     5401 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     5461 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     5521 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5581 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5641 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5701 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5761 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     5821 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5881 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5941 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6001 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6061 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6121 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6181 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6241 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6301 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
