LOCUS       sequence-213162-        5797 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 5797)
  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 5797)
  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: EYFP-IKB
FEATURES             Location/Qualifiers
     source          1..5797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        54..357
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        358..561
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     512..532
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      600..609
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(606..608,609..611,612..1322)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(651..672)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(912..931)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1259..1280
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1323..1388
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2578..2699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2615..2634)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2669..2688
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2706..3161)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2843..2864)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3055..3074
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3188..3292
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3294..3651
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3294..3485
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3317..3337)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3502..3637
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3686..4480
                     /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             3686..4480
                     /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(3740..3759)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4350..4369
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4668..4687)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4712..4759
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5088..5676
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5577..5596
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
       61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg
      601 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      661 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
      721 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
      781 ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt cgcccgctac cccgaccaca
      841 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
      901 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
      961 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1021 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     1081 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     1141 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1201 accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     1261 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1321 agtccggact cagatctcga gctcaagctt cgaattctgc agtcgacggt accgcgggcc
     1381 cgggatccat gttccagccc gcagagcccg gccaggagtg ggccatggag gggccccggg
     1441 acgcgctcaa gaaggagcgg ctactggatg accgccacga cagcggcctg gactccatga
     1501 aggacgagga gtacgagcag atggtgaagg agctgcgcga gatccgcctc gagccgcagg
     1561 aggcgccccg cggcgccgag ccctggaagc agcagctcac cgaggacgga gactcgttcc
     1621 tgcacttggc catcatccat gaagagaagg cactgaccat ggaagtggtc cgccaagtga
     1681 agggagatct ggcttttctt aacttccaga acaacctgca gcagactcca ctccacttgg
     1741 cggtgatcac caaccagcca gaaatcgctg aggcacttct ggaagctggc tgtgatcctg
     1801 agctccgaga ctttcgagga aatacccctc tacaccttgc ctgtgagcag ggctgcctgg
     1861 ccagtgtggg agtcctgact cagccccgcg ggacccagca cctccactcc attctgcagg
     1921 ccaccaacta caatggccac acatgtctgc acttagcctc gatccatggc tacctgggca
     1981 ttgtggagct gttggtgtct ttgggtgctg atgtcaacgc tcaggagccc tgcaatggcc
     2041 gaaccgccct gcatcttgcg gtggacctgc agaatcccga cctggtgtcg ctcttgttga
     2101 agtgtggggc tgatgtcaac agagtcacct accagggcta ctccccgtac cagctcacct
     2161 ggggccgccc aagcactcgg atacagcagc agctgggcca gctgacccta gaaaacctcc
     2221 agatgcttcc agagagcgag gatgaggaga gctatgacac ggagtcagag ttcacagagg
     2281 atgagctgcc ctatgacgac tgcgtgcttg gaggccagcg cctgacgtta tgagctttgg
     2341 aaagtgtcta aaagaccatg tacttgtaca tttgtacaaa atcaagagtt ttatttttct
     2401 aaaaaaaaaa aaaaaaaact cgaggggggg cccggtaccg cgggcccggg atccaccgga
     2461 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2521 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     2581 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     2641 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     2701 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     2761 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     2821 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     2881 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     2941 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3001 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3061 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3121 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     3181 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3241 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     3301 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3361 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3421 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3481 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3541 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3601 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     3661 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3721 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3781 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3841 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     3901 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3961 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4021 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4081 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4141 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4201 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4261 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4321 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4381 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4441 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4501 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4561 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4621 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     4681 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     4741 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     4801 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     4861 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     4921 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4981 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5041 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5101 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5161 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5221 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5281 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5341 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5401 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5461 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5521 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5581 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5641 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5701 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     5761 tctgtggata accgtattac cgccatgcat tagttat
//
