LOCUS       sequence-228537-        5845 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nuclear localized GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5845)
  AUTHORS   Economo MN, Viswanathan S, Tasic B, Bas E, Winnubst J, Menon V, 
            Graybuck LT, Nguyen TN, Smith KA, Yao Z, Wang L, Gerfen CR, 
            Chandrashekar J, Zeng H, Looger LL, Svoboda K
  TITLE     Distinct descending motor cortex pathways and their roles in 
            movement.
  JOURNAL   Nature. 2018 Nov;563(7729):79-84. doi: 10.1038/s41586-018-0642-9. 
            Epub 2018 Oct 31.
  PUBMED    30382200
REFERENCE   2  (bases 1 to 5845)
  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: 
            "10.1038/s41586-018-0642-9"; journalName: "Nature"; date: "2018-11";
            volume: "563"; issue: "7729"; pages: "79-84"
COMMENT     Sequence Label: pAAV-CAG-H2B-GFP
FEATURES             Location/Qualifiers
     source          1..5845
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        441..716
                     /label=chicken beta-actin promoter
     primer_bind     995..1017
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1079..1098
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1161..1538
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKENYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      1551..1560
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1557..1559,1560..1562,1563..2276)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1602..1623)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1863..1882)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2210..2231
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2289..2877
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2342..2362)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2760..2771)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2920..3041
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2931..2950)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2985..3004
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3203..3332
                     /label=AAV2 ITR
     rep_origin      complement(3569..4157)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3649..3668)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4328..5119,5120..5188))
                     /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     4951..4970
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5189..5293)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5319..5774)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5456..5477)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5668..5687
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagagta ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      241 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      301 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      361 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      421 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
      481 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
      541 gggggcgggg gggggggggg cgcgsgccag gcggggcggg gcggggcgag gggcggggcg
      601 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
      661 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
      721 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
      781 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
      841 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc
      901 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggctg tccgcggggg
      961 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     1021 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     1081 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattggatcc gaattcacta
     1141 gtctcgagga tatcgccacc atgccagagc cagcgaagtc tgctcccgcc ccgaaaaagg
     1201 gctccaagaa ggcggtgact aaggcgcaga agaaaggcgg caagaagcgc aagcgcagcc
     1261 gcaaggagaa ctattccatc tatgtgtaca aggttctgaa gcaggtccac cctgacaccg
     1321 gcatttcgtc caaggccatg ggcatcatga attcgtttgt gaacgacatt ttcgagcgca
     1381 tcgcaggtga ggcttcccgc ctggcgcatt acaacaagcg ctcgaccatc acctccaggg
     1441 agatccagac ggccgtgcgc ctgctgctgc ctggggagtt ggccaagcac gccgtgtccg
     1501 agggtactaa ggccatcacc aagtacacca gcgctaagga tccaccggtc gccaccatgg
     1561 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1621 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1681 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1741 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     1801 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1861 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1921 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1981 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     2041 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     2101 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     2161 tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     2221 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaaagc
     2281 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     2341 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     2401 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     2461 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     2521 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     2581 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     2641 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     2701 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     2761 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     2821 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     2881 gataccgtcg acccgggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg
     2941 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     3001 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     3061 attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct
     3121 acaaatgtgg taaaatcgat aaggatcttc ctagagcatg gctacgtaga taagtagcat
     3181 ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg
     3241 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     3301 cgggcggcct cagtgagcga gcgagcgcgc agctgcatta atgaatcggc caacgcgcgg
     3361 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3421 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3481 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3541 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3601 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3661 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3721 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3781 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3841 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3901 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3961 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4021 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4081 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4141 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4201 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4261 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4321 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4381 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4441 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4501 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4561 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4621 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4681 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4741 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4801 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4861 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4921 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4981 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5041 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5101 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5161 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5221 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5281 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     5341 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     5401 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     5461 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     5521 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     5581 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     5641 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     5701 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     5761 gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     5821 gcgggcctct tcgctattac gccag
//
