LOCUS       sequence-215935-        5915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express eGFP in variety of cell types.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5915)
  AUTHORS   Velez-Fort M, Bracey EF, Keshavarzi S, Rousseau CV, Cossell L, Lenzi
            SC, Strom M, Margrie TW
  TITLE     A Circuit for Integration of Head- and Visual-Motion Signals in 
            Layer 6 of Mouse Primary Visual Cortex.
  JOURNAL   Neuron. 2018 Apr 4;98(1):179-191.e6. doi: 
            10.1016/j.neuron.2018.02.023. Epub 2018 Mar 15.
  PUBMED    29551490
REFERENCE   2  (bases 1 to 5915)
  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.neuron.2018.02.023"; journalName: "Neuron"; date: 
            "2018-04-4- 4"; volume: "98"; issue: "1"; pages: "179-191.e6"
COMMENT     Sequence Label: pAAV CAGG eGFP
FEATURES             Location/Qualifiers
     source          1..5915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     enhancer        216..519
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        521..797
                     /label=chicken beta-actin promoter
     intron          798..1806
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1730..1752
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1814..1833
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(1928..1930,1931..1933,1934..2647)
                     /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             join(1928..1930,1931..1933,1934..2644)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1973..1994)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2234..2253)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2581..2602
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2662..3138
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2861..2880)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3178..3318
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3178..3307
                     /label=AAV2 ITR
     rep_origin      3393..3848
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3480..3499)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3690..3711
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3865..3884)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3984..4006
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4044..4062)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4130..4234
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4235..4303,4304..5095)
                     /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     complement(4453..4472)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5266..5854
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5755..5774
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgctagtt attaatagta atcaattacg
      181 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      241 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      301 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      361 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      421 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      481 tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc
      541 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt
      601 taattatttt gtgcagcgat gggggcgggg gggggggggg gcgcgcgcca ggcggggcgg
      661 ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg
      721 gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc
      781 gaagcgcgcg gcgggcggga gtcgctgcgt tgccttcgcc ccgtgccccg ctccgcgccg
      841 cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg
      901 acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc
      961 tgtggctgcg tgaaagcctt gaggggctcc gggagctttg tgcgggggga gcggctcggg
     1021 gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggcccgcgct gcccggcggc
     1081 tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc
     1141 gcggccgggg gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg
     1201 cggggtgtgt gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc
     1261 cccctgcacc cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt
     1321 acggggcgtg gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg
     1381 ggcggggcgg ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg
     1441 agcgccggcg gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc
     1501 gagagggcgc agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc
     1561 cgccgcaccc cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg
     1621 ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg
     1681 ggctgtccgc ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc
     1741 tggcgtgtga ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc
     1801 ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattg
     1861 cggccgtctc aggccgaatt catcgattgt ttaaacgcgg ccgcatgcat ttaaatcccg
     1921 ggatctgatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1981 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     2041 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     2101 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     2161 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     2221 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     2281 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     2341 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     2401 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     2461 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     2521 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     2581 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     2641 caagtaactc gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg
     2701 gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc
     2761 attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc
     2821 aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg
     2881 agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc
     2941 ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt
     3001 ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa
     3061 tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact
     3121 gctcccttcc ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg
     3181 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     3241 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     3301 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3361 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     3421 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3481 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3541 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3601 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3661 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3721 caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3781 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     3841 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     3901 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3961 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4021 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     4081 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4141 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4201 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4261 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4321 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4381 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4441 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4501 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4561 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4621 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4681 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4741 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4801 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4861 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4921 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4981 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5041 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5101 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5161 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5221 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5281 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5341 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5401 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5461 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5521 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5581 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5641 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5701 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5761 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5821 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5881 gttcctggcc ttttgctggc cttttgctca catgt
//
