LOCUS Exported 6621 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Human gamma-synuclein (hSncg) promoter-mediates gene expression in retinal neurons with fluorescent reporter. ACCESSION . VERSION . KEYWORDS pAM-AAV-hSncg-EGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6621) AUTHORS Wang Q, Zhuang PS, Huang H, Li L, Liu L, Webber HC, Dalal R, Siew L, Fligor CM, Chang KC, Nahmou M, Kreymerman A, Sun Y, Meyer JS, Goldberg JL, Hu Y TITLE Mouse gamma-Synuclein Promoter-Mediated Gene Expression and Editing in Mammalian Retinal Ganglion Cells. JOURNAL J Neurosci. 2020 Apr 9. pii: JNEUROSCI.0102-20.2020. doi: 10.1523/JNEUROSCI.0102-20.2020. PUBMED 32300046 REFERENCE 2 (bases 1 to 6621) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6621 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 175..193 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 175..192 /label=SP6 /note="SP6 promoter, forward primer" repeat_region 214..343 /label=AAV2 ITR CDS 1877..2596 /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 1877..2593 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1922..1943) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2183..2202) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2530..2551 /label=EGFP-C /note="EGFP, forward primer" misc_feature 2654..3242 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2707..2727) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3125..3136) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(3269..3286) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3275..3499 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" promoter complement(3709..4038) /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3723..3858 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(3777..3796) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 4023..4043 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" primer_bind complement(4113..4130) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4284..4872) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4364..4383) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5043..5903) /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 5666..5685 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5904..6008) /gene="bla" /label=AmpR promoter primer_bind 6076..6094 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(6132..6154) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 6254..6273 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6487..6506 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 ggcgttacca gaaactcaga aggttcgtcc aaccaaaccg actctgacgg cagtttacga 61 gagagatgat agggtctgct tcagtaagcc agatgctaca caattaggct tgtacatatt 121 gtcgttagaa cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag 181 gtgacactat agaatacacg gaattaattc tagctgcgcg ctcgctcgct cactgaggcc 241 gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga 301 gcgcgcagag agggagtggc caactccatc actaggggtt ccttgtagtt aatgattaac 361 ccgccatgct acttatctac gtagccatgc tctaggtacc attgacgtca ataatgacgt 421 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac 481 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaaggtcc aggccctctt 541 accgtccaac ggggtccttg ctggtgtcct cagcctctgc cttccagacc ccaggtgtcc 601 tggagctctg cagatcagag aggctagtac tggaagcctg ggcccatgcc cccagcagcc 661 cccagcccag ggggccccca aggctgaaca gcaagctcag gatcatcttg gtggtggggc 721 aggctcagct cacactcagc cttggcaagt agctccagaa actgctagtg acgttgtctt 781 caagttaaat ctcaggagga aaagaaaata cgaggacaac aaagagagga agtggcctgg 841 gccggcctac ccggtgggtc ttgtcctgcc ccccaactac cctggctggc cccacagggg 901 ccgccaacca cacaagccag ttcctgtccc tgaggacttg gctcagggac tctgggaatg 961 tggtagacat ggggtggccc caccaaatgc atccttatgg gaacctgctc cctgggagcc 1021 atgaaaagag cgtggacttc gaggtggggc cacaggaagt ggtcaggtcc atctcagggg 1081 acctgctgcc catccacact gctggccagg aaatgggggg caattcatgc ctcctcagca 1141 ccttcagcac tgggcggctc aaagaaggca agggactatt ctggggtcac acagcatgca 1201 gccagaggcc aaggcatgag gaagtccttc atttccccac ccccacccac ctcagatcct 1261 ccaaccggtt tcatggcagc ccagggtcca gcggcatcca ggatgctggt gggtagctgc 1321 acagcccagg ccgcgggagg ttggctgctc tcacctaaca ggcctatgtg gccctgaccc 1381 ctacctagga agctggggac aatggccaag gcgcctcccc tctctgtgcc tgtctgtcca 1441 ggtgcagcat agacacagca cccctggggc caagagcacc cagccagggc tgcccccatg 1501 ggtgggcagg gcagtaaatg aatgagggac aggttgggag gtggccagcc ccctccagcc 1561 catggagggc acggggcagg agagctgggc tgagccagca ggagcccagg gagcctggtc 1621 tctgccttcc tatcctggag gaaggtgagg ctgaacctcc ttccctccct ccctccctcc 1681 ccgcccccac tgcacgcagg gctggctggg ctccagctgg cctccgcatc aatatttcat 1741 cggcgtcaat aggaggcatc ggggacagcc gctgcggcag cactcgagcc agctcaagcc 1801 cgcagctcgc agggagatcc agctccgtcc tgcctgcagc agcccaaccc tgcacaccca 1861 ccatggatgt gaattcatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat 1921 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga 1981 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc 2041 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta 2101 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca 2161 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt 2221 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg 2281 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc 2341 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg 2401 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct 2461 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa 2521 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga 2581 cgagctgtac aagtaaagat ctacgcgtgg tacctctaga gtcgacccgg gcggcctcga 2641 gaagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct 2701 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc 2761 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct 2821 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga 2881 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc 2941 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac 3001 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt 3061 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt 3121 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc 3181 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc 3241 gcatcgatac cgtcgactcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct 3301 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt 3361 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg 3421 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg 3481 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc gactagagca 3541 tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac ccctagtgat 3601 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt 3661 cgcccgacgc ccgggcggcc tcagtgagcg agcgagcgcg cagagctttt tgcaaaagcc 3721 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc 3781 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg 3841 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag 3901 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc 3961 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc 4021 acaccctaac tgacacacat tccacagctg cattaatgaa tcggccaacg cgcggggaga 4081 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 4141 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 4201 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 4261 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 4321 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 4381 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 4441 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 4501 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 4561 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 4621 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 4681 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 4741 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 4801 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 4861 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 4921 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 4981 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 5041 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 5101 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 5161 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 5221 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 5281 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 5341 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 5401 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 5461 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 5521 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 5581 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 5641 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 5701 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 5761 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 5821 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 5881 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 5941 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 6001 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg 6061 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat 6121 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg 6181 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc 6241 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccattc gacgctctcc 6301 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc 6361 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc 6421 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc 6481 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc 6541 ggccacgatg cgtccggcgt agaggatctg gctagcgatg accctgctga ttggttcgct 6601 gaccatttcc gggtgcggga c //