LOCUS       sequence-383896-        4620 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses eGFP under control of short GFAP promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4620)
  AUTHORS   Bray ER, Yungher BJ, Levay K, Ribeiro M, Dvoryanchikov G, Ayupe AC, 
            Thakor K, Marks V, Randolph M, Danzi MC, Schmidt TM, Chaudhari N, 
            Lemmon VP, Hattar S, Park KK
  TITLE     Thrombospondin-1 Mediates Axon Regeneration in Retinal Ganglion 
            Cells.
  JOURNAL   Neuron. 2019 Aug 21;103(4):642-657.e7. doi: 
            10.1016/j.neuron.2019.05.044. Epub 2019 Jun 26.
  PUBMED    31255486
REFERENCE   2  (bases 1 to 4620)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.neuron.2019.05.044"; journalName: "Neuron"; date: 
            "2019-08-21- 21"; volume: "103"; issue: "4"; pages: "642-657.e7"
COMMENT     Sequence Label: pAAV.GFAP.SV40.eGFP.SV40(polyA)
FEATURES             Location/Qualifiers
     source          1..4620
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             join(979..981,982..984,985..1698)
                     /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(979..981,982..984,985..1695)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1024..1045)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1285..1304)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1632..1653
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1752..1771
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   1883..2023
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1883..2012
                     /label=AAV2 ITR
     rep_origin      2098..2553
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2185..2204)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2395..2416
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2570..2589)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2689..2711
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2749..2767)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2835..2939
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2940..3008,3009..3800)
                     /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"
     rep_origin      3971..4559
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4460..4479
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgctc gtgacctagc aacatatcct ggtgtggagt aggggacgct
      181 gctctgacag aggctcgggg gcctgagctg gctctgtgag cttgggagga ggcagacagc
      241 caggccttgt ctgcaagcag acctggcagc attgggctgg ccgcccccca gggcctcctc
      301 ttcatgccca gtgaatgact caccttggca cagacacaat gttcggggtg ggcacagtgc
      361 ctgcttcccg ccgcacccca gcccccctca aatgccttcc gagaagccca ttgagcaggg
      421 ggcttgcatt gcaccccagc ctgacagcct ggcatcttgg gataaaagca gcacagcccc
      481 ctaggggctg cccttgctgt gtggcgccac cgtcggtgga gaacaaggct ctattcagcc
      541 tgtgcccagg aaaggggatc aggggatgcc caggcatgga cagtgggtgg caggggggga
      601 gaggagggct gtctgcttcc cagaagtcca aggacacaaa tgggtgaggg gagagctctc
      661 cccatagctg ggctgcggcc caaccccacc ccctcaggct atgccagggg gtgttgccag
      721 gggcacccgg gcatcgccag tctatcccac tccttcataa agccctcgca tcccaggagc
      781 gagcagagcc agagcaggtt ggagaggaga cgcatcacct ccgctgctcg cggggtccgg
      841 actctaaggt aaatataaaa tttttaagtg tataatgtgt taaactactg attctaattg
      901 tttctctctt ttagattcca acctttggaa ctgaattccg cgggcccggg atcctctaga
      961 gtcgacctgc agaagcttat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     1021 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     1081 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     1141 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     1201 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     1261 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     1321 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     1381 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     1441 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     1501 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1561 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     1621 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     1681 gacgagctgt acaagtaact cgagagatct aggccgctag gcctcacctg cgatctcgat
     1741 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     1801 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     1861 aggtttttta aactgcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     1921 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     1981 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     2041 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     2101 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2161 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2221 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2281 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     2341 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2401 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     2461 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2521 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     2581 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     2641 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     2701 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     2761 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     2821 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     2881 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     2941 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3001 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3061 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3121 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3181 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3241 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3301 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3361 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3421 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3481 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3541 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3601 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3661 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3721 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3781 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3841 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3901 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3961 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4021 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4081 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4141 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4201 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4261 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4321 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4381 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4441 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4501 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4561 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
