LOCUS       sequence-302030-        6667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mouse Isl2 (mIsl2) promoter-mediates gene expression in retinal 
            neurons with fluorescent reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6667)
  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 6667)
  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: "J Neurosci.
            2020 Apr 9. pii: JNEUROSCI.0102-20.2020. doi: 
            10.1523/JNEUROSCI.0102-20.2020."
COMMENT     Sequence Label: pAM-AAV-mIsl2-EGPF
FEATURES             Location/Qualifiers
     source          1..6667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        121..139
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     121..138
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     repeat_region   160..289
                     /label=AAV2 ITR
     CDS             join(1869..1871,1872..1874,1875..2588)
                     /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(1869..1871,1872..1874,1875..2585)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1914..1935)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2175..2194)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2522..2543
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2646..3234
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2699..2719)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3117..3128)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3261..3278)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3267..3491
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(3701..4030)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3715..3850
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4015..4035
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4276..4864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4356..4375)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5035..5826,5827..5895))
                     /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     5658..5677
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5896..6000)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6068..6086
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6124..6146)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6246..6265
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6479..6498
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ttacgagaga gatgataggg tctgcttcag taagccagat gctacacaat taggcttgta
       61 catattgtcg ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg
      121 atttaggtga cactatagaa tacacggaat taattctagc tgcgcgctcg ctcgctcact
      181 gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc
      241 gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctt gtagttaatg
      301 attaacccgc catgctactt atctacgtag ccatgctcta ggtaccattg acgtcaataa
      361 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      421 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatggac tgcaaccatt
      481 tcccagccca gcagcttcgg ctcttagcag ttggaccgcc gcgcctgaat tagatagatc
      541 tcgggcagcg aggcctgcgg gatccaagtc cggacatagc ttttggtcga gatactttat
      601 cccagaagcc cacccgcgcc ccacttagag aggtatcaag tttttggtgt cctttcagaa
      661 tgggagtgag ggaccaacag accaccgttg gataaacact gtttgtacct catcgatttg
      721 tctttcaacc ctttgaaaca aaattcacaa cgatttttgt ttaaaaaaat cccgagtctt
      781 agaaatcgca gatttttcta ggattaaaaa gccaccggga agacggtaaa ttaccccgtg
      841 tcccacattt gtttctgatt agttgtgttg taattggttt gtgtaaccca aaatcagaga
      901 gttcgtaaaa atcttttact gtatgagtga cacatcacag cactgttttt aggctctatg
      961 cagctgagag gagaaaggag agggtgggag tcagcatcaa tttaaccaaa agagcaaaaa
     1021 gtaggaggga ggaaaagtgc ctaggccagg gtgggtggga ggactgtggc tctaagtctc
     1081 tggattcctt ttggcttacc cacctgcaac atcgagtttt ctacaggcgg cgcgggtggg
     1141 aaagtccgca tagagtttat ccaaacccag gtgctcattt cctacccgtt ctccctaact
     1201 tggaccccgc cgaacaaagg ctcaggccca ccagggagaa aggtggccgg ctggaccagt
     1261 gatggtcccg aaacagcgaa tcctcttttc tccctcaagg tctgtcgtcc ccaggaggaa
     1321 tttttaattt aaaaattata aaagaaaaac acttttaagg gggagacgac tctcatctat
     1381 tcccagaaga ctattgcctt gctaggcaaa cgccctacaa gtccgaagaa agcgcggcgc
     1441 tgcggcagag cccgcggggt ggagatgcgg cttccaaggg attgcgggac ccggggctgc
     1501 agagaggatc taggaaggtt tcgggggagg gggctgcgct ccattgttca ctccgaacca
     1561 atcagagctt tcccacttcc tcccagccaa tctccctgcg cctccgtgtc taccccagcc
     1621 caccctccgg gtccccatca tctcccgagc aagccccagg gcggtgggag agctgggtga
     1681 cgctacaggt cccgcctcca gctcgggccc cgggcgagtg cgtgctgacg tcatgctgcg
     1741 tgcgggccgg tgcggaatcg ctccttcaac tccgcggggc aggagttagt tagcaaagag
     1801 cagaggctgg gcgctcgacc ctggtccttc tgtccccggc cgcaagcttt gctcacatct
     1861 ctgaattcat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1921 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1981 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     2041 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     2101 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     2161 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     2221 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     2281 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     2341 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     2401 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     2461 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     2521 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     2581 acaagtaaag atctacgcgt ggtacctcta gagtcgaccc gggcggcctc gagaagctta
     2641 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2701 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2761 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     2821 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     2881 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     2941 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3001 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     3061 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3121 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3181 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     3241 accgtcgact cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3301 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3361 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3421 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3481 gggctctatg gcttctgagg cggaaagaac cagctggggc tcgactagag catggctacg
     3541 tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg
     3601 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     3661 gcccgggcgg cctcagtgag cgagcgagcg cgcagagctt tttgcaaaag cctaggcctc
     3721 caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg
     3781 cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta
     3841 ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc
     3901 tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat
     3961 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta
     4021 actgacacac attccacagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4081 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4141 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4201 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4261 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4321 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4381 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4441 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4501 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4561 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4621 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4681 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4741 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4801 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4861 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4921 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4981 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5041 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5101 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5161 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5221 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5281 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5341 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5401 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5461 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5521 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5581 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5641 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5701 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5761 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5821 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5881 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5941 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6001 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     6061 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     6121 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     6181 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     6241 ctatgcggca tcagagcaga ttgtactgag agtgcaccat tcgacgctct cccttatgcg
     6301 actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa
     6361 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     6421 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     6481 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     6541 tgcgtccggc gtagaggatc tggctagcga tgaccctgct gattggttcg ctgaccattt
     6601 ccgggtgcgg gacggcgtta ccagaaactc agaaggttcg tccaaccaaa ccgactctga
     6661 cggcagt
//
