LOCUS       sequence-213124-        7581 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 7581)
  AUTHORS   Birbach A, Gold P, Binder BR, Hofer E, de Martin R, Schmid JA
  TITLE     Signaling molecules of the NF-kappa B pathway shuttle constitutively
            between cytoplasm and nucleus.
  JOURNAL   J Biol Chem. 2002 Mar 29;277(13):10842-51. doi: 
            10.1074/jbc.M112475200. Epub 2002 Jan 18.
  PUBMED    11801607
REFERENCE   2  (bases 1 to 7581)
  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.M112475200"; journalName: "J Biol Chem"; date: 
            "2002-03-29- 29"; volume: "277"; issue: "13"; pages: "10842-51"
COMMENT     Sequence Label: EGFP-NIK
FEATURES             Location/Qualifiers
     source          1..7581
                     /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"
     misc_feature    1388..1453
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    4427..4548
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4464..4483)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4518..4537
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4555..5010)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4692..4713)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4904..4923
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5037..5141
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5143..5500
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5143..5334
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5166..5186)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5351..5486
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5535..6329
                     /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             5535..6329
                     /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(5589..5608)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6199..6218
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6517..6536)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6561..6608
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6937..7525
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7426..7445
                     /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 agcttcgaat tctgcagtcg acggtaccgc
     1441 gggcccggga tccaccggat ctagcatggc agtgatggaa atggcctgcc caggtgcccc
     1501 tggctcagca gtggggcagc agaaggaact ccccaaagcc aaggagaaga cgccgccact
     1561 ggggaagaaa cagagctccg tctacaagct tgaggccgtg gagaagagcc ctgtgttctg
     1621 cggaaagtgg gagatcctga atgacgtgat taccaagggc acagccaagg aaggctccga
     1681 ggcagggcca gctgccatct ctatcatcgc ccaggctgag tgtgagaata gccaagagtt
     1741 cagccccacc ttttcagaac gcattttcat cgctgggtcc aaacagtaca gccagtccga
     1801 gagtcttgat cagatcccca acaatgtggc ccatgctaca gagggcaaaa tggcccgtgt
     1861 gtgttggaag ggaaagcgtc gcagcaaagc ccggaagaaa cggaagaaga agagctcaaa
     1921 gtccctggct catgcaggag tggccttggc caaacccctc cccaggaccc ctgagcagga
     1981 gagctgcacc atcccagtgc aggaggatga gtctccactc ggcgccccat atgttagaaa
     2041 caccccgcag ttcaccaagc ctctgaagga accaggcctt gggcaactct gttttaagca
     2101 gcttggcgag ggcctacggc cggctctgcc tcgatcagaa ctccacaaac tgatcagccc
     2161 cttgcaatgt ctgaaccacg tgtggaaact gcaccacccc caggacggag gccccctgcc
     2221 cctgcccacg caccccttcc cctatagcag actgcctcat cccttcccat tccaccctct
     2281 ccagccctgg aaacctcacc ctctggagtc cttcctgggc aaactggcct gtgtagacag
     2341 ccagaaaccc ttgcctgacc cacacctgag caaactggcc tgtgtagaca gtccaaagcc
     2401 cctgcctggc ccacacctgg agcccagctg cctgtctcgt ggtgcccatg agaagttttc
     2461 tgtggaggaa tacctagtgc atgctctgca aggcagcgtg agctcaggcc aggcccacag
     2521 cctgaccagc ctggccaaga cctgggcagc aaggggctcc agatcccggg agcccagccc
     2581 caaaactgag gacaacgagg gtgtcctgct cactgagaaa ctcaagccag tggattatga
     2641 gtaccgagaa gaagtccact gggccacgca ccagctccgc ctgggcagag gctccttcgg
     2701 agaggtgcac aggatggagg acaagcagac tggcttccag tgcgctgtca aaaaggtgcg
     2761 gctggaagta tttcgggcag aggagctgat ggcatgtgca ggattgacct cacccagaat
     2821 tgtccctttg tatggagctg tgagagaagg gccttgggtc aacatcttca tggagctgct
     2881 ggaaggtggc tccctgggcc agctggtcaa ggagcagggc tgtctcccag aggaccgggc
     2941 cctgtactac ctgggccagg ccctggaggg tctggaatac ctccactcac gaaggattct
     3001 gcatggggac gtcaaagctg acaacgtgct cctgtccagc gatgggagcc acgcagccct
     3061 ctgtgacttt ggccatgctg tgtgtcttca acctgatggc ctgggaaagt ccttgctcac
     3121 aggggactac atccctggca cagagaccca catggctccg gaggtggtgc tgggcaggag
     3181 ctgcgacgcc aaggtggatg tctggagcag ctgctgtatg atgctgcaca tgctcaacgg
     3241 ctgccacccc tggactcagt tcttccgagg gccgctctgc ctcaagattg ccagcgagcc
     3301 tccgcctgtg agggagatcc caccctcctg cgcccctctc acagcccagg ccatccaaga
     3361 ggggctgagg aaagagccca tccaccgcgt gtctgcagcg gagctgggag ggaaggtgaa
     3421 ccgggcacta cagcaagtgg gaggtctgaa gagcccttgg aggggagaat ataaagaacc
     3481 aagacatcca ccgccaaatc aagccaatta ccaccagacc ctccatgccc agccgagaga
     3541 gctttcgcca agggccccag ggccccggcc agctgaggag acaacaggca gagcccctaa
     3601 gctccagcct cctctcccac cagagccccc agagccaaac aagtctcctc ccttgacttt
     3661 gagcaaggag gagtctggga tgtgggaacc cttacctctg tcctccctgg agccagcccc
     3721 tgccagaaac cccagctcac cagagcggaa agcaaccgtc ccggagcagg aactgcagca
     3781 gctggaaata gaattattcc tcaacagcct gtcccagcca ttttctctgg aggagcagga
     3841 gcaaattctc tcgtgcctca gcatcgacag cctctccctg tcggatgaca gtgagaagaa
     3901 cccatcaaag gcctctcaaa gctcgcggga caccctgagc tcaggcgtac actcctggag
     3961 cagccaggcc gaggctcgaa gctccagctg gaacatggtg ctggcccggg ggcggcccac
     4021 cgacacccca agctatttca atggtgtgaa agtccaaata cagtctctta atggtgaaca
     4081 cctgcacatc cgggagttcc accgggtcaa agtgggagac atcgccactg gcatcagcag
     4141 ccagatccca gctgcagcct tcagcttggt caccaaagac gggcagcctg ttcgctacga
     4201 catggaggtg ccagactcgg gcatcgacct gcagtgcaca ctggcccctg atggcagctt
     4261 cgcctggagc tggagggtca agcatggcca gctggagaac aggccctaat ctagataact
     4321 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     4381 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     4441 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4501 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg
     4561 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     4621 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     4681 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     4741 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     4801 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     4861 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     4921 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     4981 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     5041 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5101 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     5161 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     5221 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     5281 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     5341 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     5401 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     5461 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     5521 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     5581 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     5641 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     5701 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     5761 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5821 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5881 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5941 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     6001 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     6061 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     6121 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     6181 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     6241 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     6301 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     6361 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     6421 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     6481 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     6541 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     6601 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     6661 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     6721 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     6781 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6841 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6901 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6961 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     7021 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     7081 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     7141 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     7201 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     7261 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     7321 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7381 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     7441 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7501 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7561 cttttgctgg ccttttgctc a
//
