LOCUS       sequence-213170-        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        119..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      665..674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(671..673,674..676,677..1387)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(671..673,674..676,677..1387)
                     /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(716..737)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(977..996)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1324..1345
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2702..3073
                     /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    3205..3326
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3242..3261)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3296..3315
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3333..3788)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3470..3491)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3682..3701
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3815..3919
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3921..4278
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3921..4112
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3944..3964)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4129..4264
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4313..5107
                     /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             4313..5107
                     /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(4367..4386)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4977..4996
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5295..5314)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5339..5386
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5715..6303
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6204..6223
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      661 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
      721 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
      781 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
      841 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
      901 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
      961 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     1021 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1081 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     1141 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     1201 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1261 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     1321 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     1381 gtacaagtcc ggactcagat ctcgagctca agcttccacc atggacgaac tgttccccct
     1441 catcttcccg gcagagccag cccaggcctc tggcccctat gtggagatca ttgagcagcc
     1501 caagcagcgg ggcatgcgct tccgctacaa gtgcgagggg cgctccgcgg gcagcatccc
     1561 aggcgagagg agcacagata ccaccaagac ccaccccacc atcaagatca atggctacac
     1621 aggaccaggg acagtgcgca tctccctggt caccaaggac cctcctcacc ggcctcaccc
     1681 ccacgagctt gtaggaaagg actgccggga tggcttctat gaggctgagc tctgcccgga
     1741 ccgctgcatc cacagtttcc agaacctggg aatccagtgt gtgaagaagc gggacctgga
     1801 gcaggctatc agtcagcgca tccagaccaa caacaacccc ttccaagttc ctatagaaga
     1861 gcagcgtggg gactacgacc tgaatgctgt gcggctctgc ttccaggtga cagtgcggga
     1921 cccatcaggc aggcccctcc gcctgccgcc tgtcctttct catcccatct ttgacaatcg
     1981 tgcccccaac actgccgagc tcaagatctg ccgagtgaac cgaaactctg gcagctgcct
     2041 cggtggggat gagatcttcc tactgtgtga caaggtgcag aaagaggaca ttgaggtgta
     2101 tttcacggga ccaggctggg aggcccgagg ctccttttcg caagctgatg tgcaccgaca
     2161 agtggccatt gtgttccgga cccctcccta cgcagacccc agcctgcagg ctcctgtgcg
     2221 tgtctccatg cagctgcggc ggccttccga ccgggagctc agtgagccca tggaattcca
     2281 gtacctgcca gatacagacg atcgtcaccg gattgaggag aaacgtaaaa ggacatatga
     2341 gaccttcaag agcatcatga agaagagtcc tttcagcgga cccaccgacc cccggcctcc
     2401 acctcgacgc attgctgtgc cttcccgcag ctcagcttct gtccccaagc cagcacccca
     2461 gccctatccc tttacgtcat ccctgagcac catcaactat gatgagtttc ccaccatggt
     2521 gtttccttct gggcagatca gccaggcctc ggccttggcc ccggcccctc cccaagtcct
     2581 gccccaggct ccagcccctg cccctgctcc agccatggta tcagctctgg cccaggcccc
     2641 agcccctgtc ccagtcctag ccccaggccc tcctcaggct gtggccccac ctgcccccaa
     2701 gcccacccag gctggggaag gaacgctgtc agaggccctg ctgcagctgc agtttgatga
     2761 tgaagacctg ggggccttgc ttggcaacag cacagaccca gctgtgttca cagacctggc
     2821 atccgtcgac aactccgagt ttcagcagct gctgaaccag ggcatacctg tggcccccca
     2881 cacaactgag cccatgctga tggagtaccc tgaggctata actcgcctag tgacaggggc
     2941 ccagaggccc cccgacccag ctcctgctcc actgggggcc ccggggctcc ccaatggcct
     3001 cctttcagga gatgaagact tctcctccat tgcggacatg gacttctcag ccctgctgag
     3061 tcagatcagc tccttggatc caccggatct agataactga tcataatcag ccataccaca
     3121 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     3181 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     3241 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3301 ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa
     3361 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3421 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     3481 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     3541 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     3601 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3661 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3721 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3781 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3841 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3901 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     3961 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4021 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4081 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4141 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4201 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4261 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     4321 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4381 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4441 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     4501 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4561 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4621 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4681 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4741 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4801 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     4861 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4921 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4981 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     5041 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5101 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5161 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5221 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     5281 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     5341 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     5401 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     5461 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     5521 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     5581 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5641 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5701 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5761 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5821 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5881 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5941 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6001 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6061 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6121 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6181 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6241 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6301 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctca
//
