LOCUS       sequence-321440-        4953 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 4953)
  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 4953)
  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..4953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(212..316)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(342..797)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(479..500)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     691..710
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        871..1855
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1890..1908
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1890..1907
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             join(1968..1970,1971..1973,1974..2684)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1968..1970,1971..1973,1974..2684)
                     /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(2013..2034)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2274..2293)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2621..2642
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2730..2750
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2754..2774
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2778..2798
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2857..2991
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2881..2900)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2935..2954
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3104..3124)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3104..3122)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3143..3159)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3143..3159)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3156..3178)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3167..3183
                     /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(3191..3197,3198..3215,3216..3221))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3236..3257
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3374..3391)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3545..4133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3625..3644)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4304..4953,1..142,143..211))
                     /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     4927..4946
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
       61 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
      121 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
      181 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
      241 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
      301 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
      361 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
      421 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
      481 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
      541 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
      601 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
      661 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
      721 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
      781 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
      841 ggtgcgggcc tcttcgctat tacgccagtc gaccatagcc aattcaatat ggcgtatatg
      901 gactcatgcc aattcaatat ggtggatctg gacctgtgcc aattcaatat ggcgtatatg
      961 gactcgtgcc aattcaatat ggtggatctg gaccccagcc aattcaatat ggcggacttg
     1021 gcaccatgcc aattcaatat ggcggacttg gcactgtgcc aactggggag gggtctactt
     1081 ggcacggtgc caagtttgag gaggggtctt ggccctgtgc caagtccgcc atattgaatt
     1141 ggcatggtgc caataatggc ggccatattg gctatatgcc aggatcaata tataggcaat
     1201 atccaatatg gccctatgcc aatatggcta ttggccaggt tcaatactat gtattggccc
     1261 tatgccatat agtattccat atatgggttt tcctattgac gtagatagcc cctcccaatg
     1321 ggcggtccca tataccatat atggggcttc ctaataccgc ccatagccac tcccccattg
     1381 acgtcaatgg tctctatata tggtctttcc tattgacgtc atatgggcgg tcctattgac
     1441 gtatatggcg cctcccccat tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc
     1501 cattgacgtc aataggacca cccaccattg acgtcaatgg gatggctcat tgcccattca
     1561 tatccgttct cacgccccct attgacgtca atgacggtaa atggcccact tggcagtaca
     1621 tcaatatcta ttaatagtaa cttggcaagt acattactat tggaaggacg ccagggtaca
     1681 ttggcagtac tcccattgac gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca
     1741 tccccattga cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat
     1801 gggcggtagg cgtgcctaat gggaggtcta tataagcaat gctcgtttag ggaaccgcca
     1861 ttctgcctgg ggacgtcgga gcaagcttga tttaggtgac actatagaat acaagctact
     1921 tgttcttttt gcaggatccc atcgattcga attcaaggcc tctcgagatg gtgagcaagg
     1981 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     2041 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     2101 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     2161 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     2221 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     2281 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     2341 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     2401 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     2461 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     2521 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     2581 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     2641 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggcggt ggaagatctc
     2701 gagctcaagc ttcgaattcc cgggtcgatc caaaaaagaa gagaaaggta gatccaaaaa
     2761 agaagagaaa ggtagatcca aaaaagaaga gaaaggtata ggtaccgcgg gcccgggatc
     2821 caccggatct agaactatag tgagtcgtat tacgtagatc cagacatgat aagatacatt
     2881 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     2941 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     3001 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaattcgcg
     3061 gccgcggcgc caatgcattg ggcccggtac ccagcttttg ttccctttag tgagggttaa
     3121 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3181 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3241 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3301 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3361 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3421 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3481 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3541 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3601 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3661 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3721 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3781 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3841 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3901 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3961 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     4021 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4081 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4141 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4201 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4261 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4321 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4381 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4441 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4501 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4561 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4621 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4681 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4741 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4801 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4861 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4921 tacgggataa taccgcgcca catagcagaa ctt
//
