LOCUS       sequence-380123-        5430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express a bright green FP to label eucaryotic cell nuclei. To be 
            used in tissue-clearing methods and other fluorescent microscopy 
            methods.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5430)
  AUTHORS   Wang Z, Romanski A, Mehra V, Wang Y, Brannigan M, Campbell BC, 
            Petsko GA, Tsoulfas P, Blackmore MG
  TITLE     Brain-wide analysis of the supraspinal connectome reveals anatomical
            correlates to functional recovery after spinal injury.
  JOURNAL   Elife. 2022 Jul 15;11. pii: 76254. doi: 10.7554/eLife.76254.
  PUBMED    35838234
REFERENCE   2  (bases 1 to 5430)
  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"; journalName: "Elife."; 
            date: "2022-07-15"; volume: "11. pii: 76254. doi"; pages: " 
            10.7554/eLife.76254"
COMMENT     Sequence Label: pAAV2-CAG-3xNLS-AausFP1
FEATURES             Location/Qualifiers
     source          1..5430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(103..122)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      916..1504
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1405..1424
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1741..1870
                     /label=AAV2 ITR
     polyA_signal    2032..2153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2069..2088)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2123..2142
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    2196..2784
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2302..2313
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2711..2731
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3529..3549)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(3553..3573)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(3577..3597)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      3606..3615
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3640..3662)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3941..4217)
                     /label=chicken beta-actin promoter
     repeat_region   4528..4657
                     /label=AAV2 ITR
     rep_origin      4729..5184
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4816..4835)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5026..5047
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5210..5314
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5315..5383,5384..5430,1..745)
                     /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"
ORIGIN
        1 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
       61 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
      121 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
      181 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
      241 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
      301 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
      361 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
      421 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
      481 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
      541 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
      601 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
      661 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
      721 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
      781 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
      841 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
      901 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
      961 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1021 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     1081 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1141 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1201 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1261 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     1321 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1381 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     1441 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     1501 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     1561 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     1621 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     1681 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     1741 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
     1801 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
     1861 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
     1921 aggaagatcc ttatcgattt taccacattt gtagaggttt tacttgcttt aaaaaacctc
     1981 ccacatctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2041 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2101 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     2161 tgctcgaagc ggccgcccgg gtcgacggta tcgatgcggg gaggcggccc aaagggagat
     2221 ccgactcgtc tgagggcgaa ggcgaagacg cggaagaggc cgcagagccg gcagcaggcc
     2281 gcgggaagga aggtccgctg gattgagggc cgaagggacg tagcagaagg acgtcccgcg
     2341 cagaatccag gtggcaacac aggcgagcag ccaaggaaag gacgatgatt tccccgacaa
     2401 caccacggaa ttgtcagtgc ccaacagccg agcccctgtc cagcagcggg caaggcaggc
     2461 ggcgatgagt tccgccgtgg caatagggag ggggaaagcg aaagtcccgg aaaggagctg
     2521 acaggtggtg gcaatgcccc aaccagtggg ggttgcgtca gcaaacacag tgcacaccac
     2581 gccacgttgc ctgacaacgg gccacaactc ctcataaaga gacagcaacc aggatttata
     2641 caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag gcattaaagc
     2701 agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca atctttcaca
     2761 aattttgtaa tccagaggtt gattatcgat aagcttgata tcctcgagtt aggcgtaggc
     2821 ggtcttgcag tcgatggctc tgaacacttc caccacgttc atgtggtccc gggtttcttt
     2881 agggtccttg ctcaggatgg tgtgggcctg aatgtggtga tagtgaggga tgtccacggg
     2941 gccacctccg agaggggtat tcagctgctg gtgagcagca atgatctggc ctccgccgat
     3001 cacgtcgtgc acaatgttga tgtagcactt gatgccgttg ttctgcacgt cgggcagcac
     3061 gtagatgcag tgcgggttgt agttgaactc aagtttcttg cccaggatgt tgccgccttt
     3121 cttgaagccg ctgccgttca gggtcactct gttgtacacg gagcccttct cgtaggtcac
     3181 ttcggctctg gtcttgtaca cgccgtcgtt ctcgaaggtg atggttctct cttgcacgta
     3241 gccctcaggc atggcgctct tggggtagtc cttgatgtcg ttgggatact tggcgaagca
     3301 ctgggcgccg tatgtcactg tggtcaggat gctgctccaa ggcacaggca gttcgccggt
     3361 tgtgcagatg aagctggcct cgatctttcc ggtattggcg tcgccgtgtc ctttgcctct
     3421 gacgctgaac ttcatgcctt ccacgtcgcc ttccatttcc accacatagg ggatcttctc
     3481 gcggaacagc agggcgccat aagacatggt ggccacaggg gatcccgtca ctttccgttt
     3541 ctttttcgga tcgacttttc tcttcttttt ggggtccacc ttccgctttt tcttggggtc
     3601 gctgcccatg gtggcgaatt caagctttct agagccgccg gtcacacgcc agaagccgaa
     3661 ccccgccctg ccccgtcccc cccgaaggca gccgtccccc cgcggacagc cccgagccgc
     3721 tccccccgca caaagggccc tcccggagcc cctcaaggct ttcacgcagc cacagaaaag
     3781 aaacaagccg tcattaaacc aagcgctaat tacagcccgg aggagaaggg ccgtcccgcc
     3841 cgctcacctg tgggagtaac gcggtcagtc agagccgggg cgggcggcgc gaggcggcgg
     3901 cggagcgggg cacggggcga aggcagcgcg cagcgactcc cgcccgccgc gcgcttcgct
     3961 ttttataggg ccgccgccgc cgccgcctcg ccataaaagg aaactttcgg agcgcgccgc
     4021 tctgattggc tgccgccgca cctctccgcc tcsccccscc ccscccctcs ccccsccccs
     4081 ccccscctgg cgcgcgcccc cccccccccc cgcccccatc gctgcacaaa ataattaaaa
     4141 aataaataaa tacaaaattg ggggtgggga ggggggggag atggggagag tgaagcagaa
     4201 cgtggggctc acctcgacca tggtaatagc gatgactaat acgtagatgt actgccaagt
     4261 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     4321 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     4381 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     4441 aacatacgtc attattgacg tcaatggtac tcttcctaga gcatggctac gtagataagt
     4501 agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct
     4561 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     4621 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg gcgtaatagc gaagaggccc
     4681 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     4741 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4801 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4861 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4921 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4981 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     5041 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     5101 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5161 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5221 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5281 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     5341 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5401 gaaagtaaaa gatgctgaag atcagttggg
//
