LOCUS       sequence-325977-        4954 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses nuclear localized EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4954)
  AUTHORS   Enyedi B, Jelcic M, Niethammer P
  TITLE     The Cell Nucleus Serves as a Mechanotransducer of Tissue 
            Damage-Induced Inflammation.
  JOURNAL   Cell. 2016 May 19;165(5):1160-1170. doi: 10.1016/j.cell.2016.04.016.
  PUBMED    27203112
REFERENCE   2  (bases 1 to 4954)
  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: 
            "10.1016/j.cell.2016.04"; journalName: "Cell"; date: "2016-05-19- 
            19"; volume: "165"; issue: "5"; pages: "1160-1170"
COMMENT     Sequence Label: pCS2+ EGFP-3NLS
FEATURES             Location/Qualifiers
     source          1..4954
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        336..354
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     336..353
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      412..421
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(418..420,421..423,424..1134)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(418..420,421..423,424..1134)
                     /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(463..484)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(724..743)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1071..1092
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1180..1200
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1204..1224
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1228..1248
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1307..1441
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1331..1350)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1385..1404
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1554..1574)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1554..1572)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1593..1609)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1593..1609)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1606..1628)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1617..1633
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1641..1647,1648..1665,1666..1671))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1686..1707
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1824..1841)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1995..2583)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2075..2094)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2754..3545,3546..3614))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3377..3396
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3615..3719)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3745..4200)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3882..3903)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4094..4113
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(4271..4954,1..301)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cattcatatc cgttctcacg ccccctattg acgtcaatga cggtaaatgg cccacttggc
       61 agtacatcaa tatctattaa tagtaacttg gcaagtacat tactattgga agtacgccag
      121 ggtacattgg cagtactccc attgacgtca atggcggtaa atggcccgcg atggctgcca
      181 agtacatccc cattgacgtc aatggggagg ggcaatgacg caaatgggcg ttccattgac
      241 gtaaatgggc ggtaggcgtg cctaatggga ggtctatata agcaatgctc gtttagggaa
      301 ccgccattct gcctggggac gtcggagcaa gcttgattta ggtgacacta tagaatacaa
      361 gctacttgtt ctttttgcag gatcccatcg atctggctag cgctaccggt cgccaccatg
      421 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      481 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      541 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      601 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
      661 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      721 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
      781 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
      841 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
      901 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
      961 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1021 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1081 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggcggt
     1141 ggaagatctc gagctcaagc ttcgaattcc cgggtcgatc caaaaaagaa gagaaaggta
     1201 gatccaaaaa agaagagaaa ggtagatcca aaaaagaaga gaaaggtata ggtaccgcgg
     1261 gcccgggatc caccggatct agaactatag tgagtcgtat tacgtagatc cagacatgat
     1321 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1381 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     1441 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     1501 ttaattcgcg gccgcggcgc caatgcattg ggcccggtac ccagcttttg ttccctttag
     1561 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     1621 tatccgctca caattccaca caacatacga gccgggagca taaagtgtaa agcctggggt
     1681 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     1741 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1801 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1861 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1921 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1981 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2041 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2101 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2161 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2221 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2281 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2341 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2401 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2461 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2521 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2581 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2641 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2701 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2761 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2821 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     2881 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     2941 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3001 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3061 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3121 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3181 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3241 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3301 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3361 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3421 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3481 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3541 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3601 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3661 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3721 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     3781 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     3841 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     3901 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     3961 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     4021 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     4081 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     4141 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     4201 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     4261 tattacgcca gtcgaccgcc aattcaatat ggcgtatatg gactcatgcc aattcaatat
     4321 ggtggatctg gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc aattcaatat
     4381 ggtggatctg gaccccagcc aattcaatat ggcggacttg gcaccatgcc aattcaatat
     4441 ggcggacctg gcactgtgcc aactggggag gggtctactt ggcacggtgc caagtttgag
     4501 gaggggtctt ggccctgtgc caagtccgcc atattgaatt ggcatggtgc caataatggc
     4561 ggccatattg gctatatgcc aggatcaata tataggcaat atccaatatg gccctatgcc
     4621 aatatggcta ttggccaggt tcaatactat gtattggccc tatgccatat agtattccat
     4681 atatgggttt tcctattgac gtagatagcc cctcccaatg ggcggtccca tataccatat
     4741 atggggcttc ctaataccgc ccatagccac tcccccattg acgtcaatgg tctctatata
     4801 tggtctttcc tattgacgtc atatgggcgg tcctattgac gtatatggcg cctcccccat
     4861 tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc aataggacca
     4921 cccaccattg acgtcaatgg gatggctcat tgcc
//
