LOCUS       Exported                6960 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  fluorescent fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    IKK2-EGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6960)
  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 6960)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6960
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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      2893..2902
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2899..3618
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2899..3615
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2944..2965)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3205..3224)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3552..3573
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3741..3862
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3778..3797)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3832..3851
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3869..4324)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4006..4027)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4218..4237
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4351..4455
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4457..4814
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4480..4500)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4665..4800
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4727..4746
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4849..5643
                     /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(4903..4922)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5513..5532
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5831..5850)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5875..5922
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6251..6839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6740..6759
                     /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 tccgctagac tcgacttcga
      601 tatctcgaat tcgatatgag ctggtcacct tccctgacaa cgcagacatg tggggcctgg
      661 gaaatgaaag agcgccttgg gacaggggga tttggaaatg tcatccgatg gcacaatcag
      721 gaaacaggtg agcagattgc catcaagcag tgccggcagg agctcagccc ccggaaccga
      781 gagcggtggt gcctggagat ccagatcatg agaaggctga cccaccccaa tgtggtggct
      841 gcccgagatg tccctgaggg gatgcagaac ttggcgccca atgacctgcc cctgctggcc
      901 atggagtact gccaaggagg agatctccgg aagtacctga accagtttga gaactgctgt
      961 ggtctgcggg aaggtgccat cctcaccttg ctgagtgaca ttgcctctgc gcttagatac
     1021 cttcatgaaa acagaatcat ccatcgggat ctaaagccag aaaacatcgt cctgcagcaa
     1081 ggagaacaga ggttaataca caaaattatt gacctaggat atgccaagga gctggatcag
     1141 ggcagtcttt gcacatcatt cgtggggacc ctgcagtacc tggccccaga gctactggag
     1201 cagcagaagt acacagtgac cgtcgactac tggagcttcg gcaccctggc ctttgagtgc
     1261 atcacgggct tccggccctt cctccccaac tggcagcccg tgcagtggca ttcaaaagtg
     1321 cggcagaaga gtgaggtgga cattgttgtt agcgaagact tgaatggaac ggtgaagttt
     1381 tcaagctctt taccctaccc caataatctt aacagtgtcc tggctgagcg actggagaag
     1441 tggctgcaac tgatgctgat gtggcacccc cgacagaggg gcacggatcc cacgtatggg
     1501 cccaatggct gcttcaaggc cctggatgac atcttaaact taaagctggt tcatatcttg
     1561 aacatggtca cgggcaccat ccacacctac cctgtgacag aggatgagag tctgcagagc
     1621 ttgaaggcca gaatccaaca ggacacgggc atcccagagg aggaccagga gctgctgcag
     1681 gaagcgggcc tggcgttgat ccccgataag cctgccactc agtgtatttc agacggcaag
     1741 ttaaatgagg gccacacatt ggacatggat cttgtttttc tctttgacaa cagtaaaatc
     1801 acctatgaga ctcagatctc cccacggccc caacctgaaa gtgtcagctg tatccttcaa
     1861 gagcccaaga ggaatctcgc cttcttccag ctgaggaagg tgtggggcca ggtctggcac
     1921 agcatccaga ccctgaagga agattgcaac cggctgcagc agggacagcg agccgccatg
     1981 atgaatctcc tccgaaacaa cagctgcctc tccaaaatga agaattccat ggcctccatg
     2041 tctcagcagc tcaaggccaa gttggatttc ttcaaaacca gcatccagat tgacctggag
     2101 aagtacagcg agcaaaccga gtttgggatc acatcagata aactgctgct ggcctggagg
     2161 gaaatggagc aggctgtgga gctctgtggg cgggagaacg aagtgaaact cctggtagaa
     2221 cggatgatgg ctctgcagac cgacattgtg gacttacaga ggagtcccat gggccggaag
     2281 caggggggaa cgctggacga cctagaggag caagcaaggg agctgtacag gagactaagg
     2341 gaaaaacctc gagaccagcg aactgagggt gacagtcagg aaatggtacg gctgctgctt
     2401 caggcaattc agagcttcga gaagaaggtg cgagtgatct atacgcagct cagtaaaact
     2461 gtggtttgca agcagaaggc gctggaactg ttgcccaagg tggaagaggt ggtgagctta
     2521 atgaatgagg atgagaagac tgttgtccgg ctgcaggaga agcggcagaa ggagctctgg
     2581 aatctcctga agattgcttg tagcaaggtc cgtggtcctg tcagtggaag cccggatagc
     2641 atgaatgcct ctcgacttag ccagcctggg cagctgatgt ctcagccctc cacggcctcc
     2701 aacagcttac ctgagccagc caagaagagt gaagaactgg tggctgaagc acataacctc
     2761 tgcaccctgc tagaaaatgc catacaggac actgtgaggg aacaagacca gagtttcacg
     2821 gccctagact ggagctggtt acagacggaa gaagaagagc acagctgcct ggagcagggg
     2881 gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     2941 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     3001 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     3061 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     3121 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     3181 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     3241 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     3301 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     3361 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     3421 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     3481 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     3541 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     3601 gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg
     3661 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     3721 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     3781 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3841 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc
     3901 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     3961 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     4021 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     4081 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     4141 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     4201 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     4261 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     4321 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4381 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4441 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     4501 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4561 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4621 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4681 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4741 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4801 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4861 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4921 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4981 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     5041 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5101 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5161 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5221 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5281 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5341 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     5401 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5461 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5521 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5581 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5641 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     5701 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5761 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     5821 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     5881 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     5941 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     6001 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     6061 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     6121 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6181 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6241 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6301 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6361 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6421 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6481 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6541 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6601 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6661 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6721 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6781 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6841 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6901 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat
//
