LOCUS       sequence-213136-        5796 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 5796)
  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 5796)
  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: ECFP-IKB
FEATURES             Location/Qualifiers
     source          1..5796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(114..702)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(194..213)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    1031..1078
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1103..1122
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(1310..2104)
                     /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(1310..2104)
                     /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(1421..1440)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2031..2050
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2139..2496)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2153..2288
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        2305..2496
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     2453..2473
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(2498..2602)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2629..3084
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2716..2735)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2926..2947
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    3091..3212
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3102..3121)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3156..3175
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    4402..4467
                     /label=MCS
                     /note="multiple cloning site"
     CDS             complement(join(4468..5178,5179..5181,5182..5184))
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4510..4531)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4859..4878
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5118..5139
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      5181..5190
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5229..5432)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5258..5278)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5433..5736
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgcatggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
       61 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      121 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      181 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      241 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      301 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      361 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      421 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      481 attagcagag cgaggtatgt aggcggtgat acagagttct tgaagtggtg gcctaactac
      541 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      601 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      661 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      721 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      781 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      841 taaagtatat atgagtaacc tgaggctatg gcagggcctg ccgccccgac gttggctgcg
      901 agccctgggc cttcacccga acttgggggg tggggtgggg aaaaggaaga aacgcgggcg
      961 tattggcccc aatggggtct cggtggggta tcgacagagt gccagccctg ggaccgaacc
     1021 ccgcgtttat gaacaaacga cccaacaccg tgcgttttat tctgtctttt tattgccgtc
     1081 atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct ccccctaggg
     1141 tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg
     1201 aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtgat
     1261 ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct cagaagaact
     1321 cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca
     1381 cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg
     1441 ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc
     1501 ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct
     1561 cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat
     1621 gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct
     1681 cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc
     1741 gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga
     1801 gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt
     1861 cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt
     1921 cttgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct
     1981 gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat
     2041 agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa
     2101 tcatgcgaaa cgatcctcat cctgtctctt gatcgatctt tgcaaaagcc taggcctcca
     2161 aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca
     2221 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg
     2281 ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt
     2341 tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg
     2401 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac
     2461 tgacacacat tccacagctg gttctttccg cctcaggact cttccttttt caatattatt
     2521 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     2581 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta
     2641 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2701 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2761 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2821 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     2881 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2941 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3001 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3061 acaaaatatt aacgcttaca atttacgcct taagatacat tgatgagttt ggacaaacca
     3121 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     3181 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     3241 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     3301 gtatggctga ttatgatcag ttatctagat ccggtggatc ccgggcccgc ggtaccgggc
     3361 cccccctcga gttttttttt ttttttttta gaaaaataaa actcttgatt ttgtacaaat
     3421 gtacaagtac atggtctttt agacactttc caaagctcat aacgtcaggc gctggcctcc
     3481 aagcacgcag tcgtcatagg gcagctcatc ctctgtgaac tctgactccg tgtcatagct
     3541 ctcctcatcc tcgctctctg gaagcatctg gaggttttct agggtcagct ggcccagctg
     3601 ctgctgtatc cgagtgcttg ggcggcccca ggtgagctgg tacggggagt agccctggta
     3661 ggtgactctg ttgacatcag ccccacactt caacaagagc gacaccaggt cgggattctg
     3721 caggtccacc gcaagatgca gggcggttcg gccattgcag ggctcctgag cgttgacatc
     3781 agcacccaaa gacaccaaca gctccacaat gcccaggtag ccatggatcg aggctaagtg
     3841 cagacatgtg tggccattgt agttggtggc ctgcagaatg gagtggaggt gctgggtccc
     3901 gcggggctga gtcaggactc ccacactggc caggcagccc tgctcacagg caaggtgtag
     3961 aggggtattt cctcgaaagt ctcggagctc aggatcacag ccagcttcca gaagtgcctc
     4021 agcgatttct ggctggttgg tgatcaccgc caagtggagt ggagtctgct gcaggttgtt
     4081 ctggaagtta agaaaagcca gatctccctt cacttggcgg accacttcca tggtcagtgc
     4141 cttctcttca tggatgatgg ccaagtgcag gaacgagtct ccgtcctcgg tgagctgctg
     4201 cttccagggc tcggcgccgc ggggcgcctc ctgcggctcg aggcggatct cgcgcagctc
     4261 cttcaccatc tgctcgtact cctcgtcctt catggagtcc aggccgctgt cgtggcggtc
     4321 atccagtagc cgctccttct tgagcgcgtc ccggggcccc tccatggccc actcctggcc
     4381 gggctctgcg ggctggaaca tggatcccgg gcccgcggta ccgtcgactg cagaattcga
     4441 agcttgagct cgagatctga gtccggactt gtacagctcg tccatgccga gagtgatccc
     4501 ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct
     4561 cagggcggac tgggtgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat
     4621 gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg
     4681 gatcttgaag ttggccttga tgccgttctt ctgcttgtcg gcggtgatat agacgttgtg
     4741 gctgatgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat
     4801 gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt
     4861 cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat
     4921 ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac
     4981 gccccaggtc agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca
     5041 gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct
     5101 gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa
     5161 cagctcctcg cccttgctca ccatggtggc gaccggtagc gctagcggat ctgacggttc
     5221 actaaaccag ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc
     5281 aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa
     5341 ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca
     5401 cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag
     5461 atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc
     5521 catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact
     5581 gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt
     5641 ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt
     5701 cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga
     5761 actaatgacc ccgtaattga ttactattaa taacta
//
