LOCUS       sequence-302081-        6658 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human Isl2 (hIsl2) 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 6658)
  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 6658)
  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-hIsl2-EGFP
FEATURES             Location/Qualifiers
     source          1..6658
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(208..227)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(441..460)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     560..582
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(620..638)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        706..810
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(811..879,880..1671)
                     /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(1029..1048)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1842..2430
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2331..2350
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2671..2691)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2676..3005
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2856..2991
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3215..3439
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3428..3445
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     misc_feature    3472..4060
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3578..3589
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3987..4007
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(4118..4831,4832..4834,4835..4837))
                     /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             complement(join(4121..4831,4832..4834,4835..4837))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4163..4184)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4512..4531
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4771..4792
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     repeat_region   6408..6537
                     /label=AAV2 ITR
     promoter        complement(6558..6576)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6559..6576)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ctggcttact gaagcagacc ctatcatctc tctcgtaaac tgccgtcaga gtcggtttgg
       61 ttggacgaac cttctgagtt tctggtaacg ccgtcccgca cccggaaatg gtcagcgaac
      121 caatcagcag ggtcatcgct agccagatcc tctacgccgg acgcatcgtg gccggcatca
      181 ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc
      241 gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg
      301 tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc
      361 tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc
      421 gtcgaatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
      481 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
      541 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
      601 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
      661 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
      721 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
      781 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
      841 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
      901 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
      961 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     1021 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     1081 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     1141 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     1201 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     1261 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     1321 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     1381 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     1441 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     1501 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     1561 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     1621 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     1681 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     1741 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     1801 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1861 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1921 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1981 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2041 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2101 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2161 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2221 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2281 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2341 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2401 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2461 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     2521 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     2581 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     2641 gcgcgttggc cgattcatta atgcagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     2701 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2761 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2821 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     2881 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     2941 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3001 gcaaaaagct ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg
     3061 gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta
     3121 ggggttcctt gtagttaatg attaacccgc catgctactt atctacgtag ccatgctcta
     3181 gtcgagcccc agctggttct ttccgcctca gaagccatag agcccaccgc atccccagca
     3241 tgcctgctat tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga
     3301 atagaatgac acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca
     3361 gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc
     3421 tggcaactag aaggcacagt cgaggctgat cagcgagtcg acggtatcga tgcggggagg
     3481 cggcccaaag ggagatccga ctcgtctgag ggcgaaggcg aagacgcgga agaggccgca
     3541 gagccggcag caggccgcgg gaaggaaggt ccgctggatt gagggccgaa gggacgtagc
     3601 agaaggacgt cccgcgcaga atccaggtgg caacacaggc gagcagccaa ggaaaggacg
     3661 atgatttccc cgacaacacc acggaattgt cagtgcccaa cagccgagcc cctgtccagc
     3721 agcgggcaag gcaggcggcg atgagttccg ccgtggcaat agggaggggg aaagcgaaag
     3781 tcccggaaag gagctgacag gtggtggcaa tgccccaacc agtgggggtt gcgtcagcaa
     3841 acacagtgca caccacgcca cgttgcctga caacgggcca caactcctca taaagagaca
     3901 gcaaccagga tttatacaag gaggagaaaa tgaaagccat acgggaagca atagcatgat
     3961 acaaaggcat taaagcagcg tatccacata gcgtaaaagg agcaacatag ttaagaatac
     4021 cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt atcgataagc ttctcgaggc
     4081 cgcccgggtc gactctagag gtaccacgcg tagatcttta cttgtacagc tcgtccatgc
     4141 cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt
     4201 tggggtcttt gctcagggcg gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg
     4261 ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg ctgccgtcct
     4321 cgatgttgtg gcggatcttg aagttcacct tgatgccgtt cttctgcttg tcggccatga
     4381 tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct
     4441 ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca
     4501 cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt
     4561 agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc
     4621 tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc acgggcagct
     4681 tgccggtggt gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcgccct
     4741 cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca
     4801 ccaccccggt gaacagctcc tcgcccttgc tcaccatgaa ttcaagagat gtgcgcaaag
     4861 tgtgcggcca ggggcagaag gacgagggtc gcgcgcccgg cctcggctct ttgctaacta
     4921 actcctactg ccccgcggag ttgaaggagc gattccgcac cggcccgcac gcagcatgac
     4981 gtcaacacgc actagccccg ggccggagct gggggcggga cctgtggcgt cacgaagctt
     5041 ccccagaact gccggggatg cgggggaggt gatggggatc caggggctga tgggcggggt
     5101 gggttgggtt ggaggctggg ggtgaaggga gattggctgg gaggaagtgg gccaaccctg
     5161 attggcccgg agtgaacaat ggagtgcagc cccccccacc tccgaaacct acaaggatcc
     5221 tctctcatgc cccgggcctg gcaatctgga ctggaccttg cgtcaccgcc gagctggctc
     5281 cgcggccgtt acaactctga gcttcctctt ggactttcgg ggcggttttc ccaggcgcgc
     5341 tggagctggg tgatggatac ctgagggtgg agatggctct cctttaagag ctcccttctt
     5401 atataaatta aaaaaaaaaa aattaaaccc actctcagga cgagagagcc gaggacaagg
     5461 gggatccgtt tggacgtgac cgtcatcgtc cagctggtca cctttctcct ctcatgtgcc
     5521 tttgttcttc gggtccaagc tagagagagt gtagagaatg accacctggg ttctgataaa
     5581 ctctatttag actttcctat ccacacacac gtagaaaact tcagattggg gcattacgtg
     5641 ggttgagcca gaacccagat ccaaaagcgc tcacccggga ctgggacggg tcctgactcc
     5701 atccttgctg ctgctgccgc tgctgctgcc cctcctccct ctcctccacc accacctcct
     5761 cctcctgctc cacttccttt tttttgttgt tttgttttgc ttttaatttg ttgctgttgt
     5821 tgttttaaat gtaacatgga cttccccgcc ccctcatttt tcccccatcc aaatggcata
     5881 gctcctttta ggaacactgc tggggtcgat tgcgtgtcca acttccaaaa taaaagattt
     5941 ttctaaactc ttgtccttca ttaacaaacc cactgcaatg caaataatca gaaacgaacg
     6001 taaggcgctg ggtaatttac agtggtatct gcgaggcggc ggctcgttgg tcctgagaaa
     6061 atctgagagc tctgagcaca gtccaggcat tctttcaaag aaatcgctgt gggttttgtt
     6121 tgaaagggtt caaagaccgg cccatgaatt ataaataaca tttatccaac catggtgtct
     6181 gttcttgttt tcccattaaa aggagtacca agaacttggt gtgcgcctgc atatgataca
     6241 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga
     6301 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat ggtacctaga
     6361 gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt
     6421 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     6481 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagcta
     6541 gaattaattc cgtgtattct atagtgtcac ctaaatcgta tgtgtatgat acataaggtt
     6601 atgtattaat tgtagccgcg ttctaacgac aatatgtaca agcctaattg tgtagcat
//
