LOCUS       sequence-465276-        5661 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP in OFF cone bipolar cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5661)
  AUTHORS   Aviles EC, Wang SK, Patel S, Cordero S, Shi S, Lin L, Kefalov VJ, 
            Goodrich LV, Cepko CL, Xue Y
  TITLE     ERG responses to high-frequency flickers require FAT3 signaling in 
            mouse retinal bipolar cells.
  JOURNAL   J Gen Physiol. 2025 Mar 3;157(2):e202413642. doi: 
            10.1085/jgp.202413642. Epub 2025 Feb 4.
  PUBMED    39903280
REFERENCE   2  (bases 1 to 5661)
  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 Gen 
            Physiol."; date: "2025-03-3"; volume: "157(2):e202413642. doi"; 
            pages: " 10.1085/jgp.202413642. Epub 2025 Feb 4"
COMMENT     Sequence Label: AAV-Grik1-GFP
FEATURES             Location/Qualifiers
     source          1..5661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   259..388
                     /label=AAV2 ITR
     intron          1237..1369
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1352..1371
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1380..1389
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1386..1388,1389..1391,1392..2105)
                     /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(1386..1388,1389..1391,1392..2102)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1431..1452)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1692..1711)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2039..2060
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2172..2192)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2590..2601)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2668..2685)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2674..2881
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2969..3098
                     /label=AAV2 ITR
     primer_bind     complement(3117..3134)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3117..3133)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3126..3148)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3275..3730
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3362..3381)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3572..3593
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3756..3860
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3861..3929,3930..4721)
                     /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(4079..4098)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4892..5480
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5381..5400
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccagattta
      241 attaaggcct taattaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
      301 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      361 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
      421 tctacgtagc catgctctag gaagatcgga attcgccctt aagctagcat gcaatttgcc
      481 ttggagatat gaccaataat caggttgatt cctcttccag gaaaggatca acccttaggt
      541 aatagcctgg tggtgtttca gtaaaaatta ggaaggaaaa ccggaagtgc attggcatct
      601 cacaccctcc attatgtaaa ctactcagtg cgcatatttt agggtatgct gtaattacag
      661 atggtctctt tctgttgtac ttggaaatga tgccctgaga gagagtgatg agttttcagg
      721 ttcttatccc tgaaccatgc tgttaccatc actctgcatt aaggccttcg tgtctccaca
      781 cccccccccc cagatccatc ctcctttaat ctcttaatct tcttttttaa tttgttccca
      841 tctgcaattt tctataattc caaaataagc ctaagtagac aactttattc gaaagggact
      901 taggaatttg cttcggatga agctgaatga aacaaattaa attatttaag aaaattattc
      961 cctactgctt attttaaggc accctaagtc ataagagaaa ggatgtgaat atgaatagcc
     1021 atacttaaca tttacagaaa tggtaagaag caggcaatgt tttcatgtta aattaattaa
     1081 aatcaatatg attcagacct acctactaga taatctatta atcagtttga aatagaaatc
     1141 ttggaacaca tgactttagt aatgttcaga aattatcctc tacataaaac tgggtgagac
     1201 agatcctgca gaagttggtc gtgaggcact gggcaggtaa gtatcaaggt tacaagacag
     1261 gtttaaggag accaatagaa actgggcttg tcgagacaga gaagactctt gcgtttctga
     1321 taggcaccta ttggtcttac tgacatccac tttgcctttc tctccacagg tgtccagcgg
     1381 ccgccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1441 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     1501 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     1561 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     1621 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1681 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1741 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1801 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     1861 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     1921 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1981 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     2041 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     2101 agtaataagc ttggatccaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2161 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2221 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     2281 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     2341 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     2401 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     2461 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     2521 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     2581 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     2641 cggcctcttc cgcgtcttcg agatctgcct cgactgtgcc ttctagttgc cagccatctg
     2701 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     2761 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     2821 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     2881 actcgagtta agggcgaatt cccgataagg atcttcctag agcatggcta cgtagataag
     2941 tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc
     3001 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     3061 tttgcccggg cggcctcagt gagcgagcga gcgcgcagcc ttaattaacc taattcactg
     3121 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     3181 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     3241 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     3301 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3361 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3421 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3481 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3541 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3601 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3661 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     3721 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3781 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3841 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3901 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3961 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4021 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4081 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4141 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4201 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4261 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4321 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4381 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     4441 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4501 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4561 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4621 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4681 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     4741 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4801 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4861 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4921 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4981 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5041 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5101 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5161 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5221 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5281 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5341 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5401 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5461 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5521 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5581 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     5641 cagtgagcga ggaagcggaa g
//
