LOCUS       sequence-324112-        7300 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid expressing full-length CX3CL1 (fractalkine) in 
            photoreceptors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7300)
  AUTHORS   Wang SK, Xue Y, Rana P, Hong CM, Cepko CL
  TITLE     Soluble CX3CL1 gene therapy improves cone survival and function in 
            mouse models of retinitis pigmentosa.
  JOURNAL   Proc Natl Acad Sci U S A. 2019 May 14;116(20):10140-10149. doi: 
            10.1073/pnas.1901787116. Epub 2019 Apr 29.
  PUBMED    31036641
REFERENCE   2  (bases 1 to 7300)
  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"; doi: 
            "10.1073/pnas.1901787116"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2019-05-14- 14"; volume: "116"; issue: "20"; pages: 
            "10140-10149"
COMMENT     Sequence Label: AAV-hRedOpsin-fCX3CL1
FEATURES             Location/Qualifiers
     source          1..7300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2214..2223
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3467..3487)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3885..3896)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3963..3980)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3969..4176
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4264..4393
                     /label=AAV2 ITR
     primer_bind     complement(4431..4447)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4431..4447)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4444..4466)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4455..4471
                     /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)."
     promoter        complement(join(4479..4485,4486..4503,4504..4509))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4524..4545
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4833..5421)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4913..4932)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5592..6383,6384..6452))
                     /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     6215..6234
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6453..6557)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6583..7038)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6720..6741)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6932..6951
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7165..7187
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7179..7196
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7180..7196
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
       61 ggttccttgt agttaatgat taacccgcca tgctacttat ctacgtagcc atgctctagg
      121 aagatcggaa ttcgccctta agctagccta cagcagccag ggtgagatta tgaggctgag
      181 ctgagaatat caagactgta ccgagtaggg ggccttggca agtgtggaga gcccggcagc
      241 tggggcagag ggcggagtac ggtgtgcgtt tacggacctc ttcaaacgag gtaggaaggt
      301 cagaagtcaa aaagggaaca aatgatgttt aaccacacaa aaatgaaaat ccaatggttg
      361 gatatccatt ccaaatacac aaaggcaacg gataagtgat ccgggccagg cacagaaggc
      421 catgcacccg taggattgca ctcagagctc ccaaatgcat aggaatagaa gggtgggtgc
      481 aggaggctga ggggtgggga aagggcatgg gtgtttcatg aggacagagc ttccgtttca
      541 tgcaatgaaa agagtttgga gacggatggt ggtgactgga ctatacactt acacacggta
      601 gcgatggtac actttgtatt atgtatattt taccacgatc tttttaaagt gtcaaaggca
      661 aatggccaaa tggttccttg tcctatagct gtagcagcca tcggctgtta gtgacaaagc
      721 ccctgagtca agatgacagc agcccccata actcctaatc ggctctcccg cgtggagtca
      781 tttaggagta gtcgcattag agacaagtcc aacatctaat cttccaccct ggccagggcc
      841 ccagctggca gcgagggtgg gagactccgg gcagagcaga gggcgctgac attggggccc
      901 ggcctggctt gggtccctct ggcctttccc caggggccct ctttccttgg ggctttcttg
      961 ggccgccact gctcccgctc ctctcccccc atcccacccc ctcaccccct cgttcttcat
     1021 atccttctct agtgctccct ccactttcat ccacccttct gcaagagtgt gggaccacaa
     1081 atgagttttc acctggcctg gggacacacg tgcccccaca ggtgctgagt gactttctag
     1141 gacagtaatc tgctttaggc taaaatggga cttgatcttc tgttagccct aatcatcaat
     1201 tagcagagcc ggtgaaggtg cagaacctac cgcctttcca ggcctcctcc cacctctgcc
     1261 acctccactc tccttcctgg gatgtggggg ctggcacacg tgtggcccag ggcattggtg
     1321 ggattgcact gagctgggtc attagcgtaa tcctggacaa gggcagacag ggcgagcgga
     1381 gggccagctc cggggctcag gcaaggctgg gggcttcccc cagacacccc actcctcctc
     1441 tgctggaccc ccacttcata gggcacttcg tgttctcaaa gggcttccaa atagcatggt
     1501 ggccttggat gcccagggaa gcctcagagt tgcttatctc cctctagaca gaaggggaat
     1561 ctcggtcaag agggagaggt cgccctgttc aaggccaccc agccagctca tggcggtaat
     1621 gggacaaggc tggccagcca tcccaccctc agaagggacc cggtggggca ggtgatctca
     1681 gaggaggctc acttctgggt ctcacattct tggatccggt tccaggcctc ggccctaaat
     1741 agtctccctg ggctttcaag agaaccacat gagaaaggag gattcgggct ctgagcagtt
     1801 tcaccaccca ccccccagtc tgcaaatcct gacccgtggg tccacctgcc ccaaaggcgg
     1861 acgcaggaca gtagaaggga acagagaaca cataaacaca gagagggcca cagcggctcc
     1921 cacagtcacc gccaccttcc tggcggggat gggtggggcg tctgagtttg gttcccagca
     1981 aatccctctg agccgccctt gcgggctcgc ctcaggagca ggggagcaag aggtgggagg
     2041 aggaggtcta agtcccaggc ccaattaaga gatcaggtag tgtagggttt gggagctttt
     2101 aaggtgaaga ggcccgggct gatcccacag gccagtataa agcgccgtga ccctcaggtg
     2161 atgcgccagg gccggctgcc gtcggggaca gggctttcca tagcggccgc gccgccacca
     2221 tggctccctc gccgctcgcg tggctgctgc gcctggccgc gttcttccat ttgtgtactc
     2281 tgctgccggg tcagcacctc ggcatgacga aatgcgaaat catgtgcgac aagatgacct
     2341 cacgaatccc agtggctttg ctcatccgct atcagctaaa ccaggagtcc tgcggcaagc
     2401 gtgccattgt cctggagacg acacagcaca gacgcttctg tgctgacccg aaggagaaat
     2461 gggtccaaga cgccatgaag catctggatc accaggctgc tgccctcact aaaaatggtg
     2521 gcaagtttga gaagcgggtg gacaatgtga cacctgggat caccttggcc actaggggac
     2581 tgtccccatc tgccctgaca aagcctgaat ccgccacatt ggaagacctt gctttggaac
     2641 tgactactat ttcccaggag gccaggggga ccatggggac ttcccaagag ccaccggcag
     2701 cagtgaccgg atcatctctc tcaacttccg aggcacagga tgcagggctt acggctaagc
     2761 ctcagagcat tggaagtttt gaggcggctg acatctccac caccgtttgg ccgagtcctg
     2821 ctgtctacca atctggatct agctcctggg ctgaggaaaa agctactgag tccccctcca
     2881 ctacagcccc atctcctcag gtgtccacta cttcaccttc aaccccagag gaaaatgttg
     2941 ggtccgaagg ccaaccccca tgggtccagg gacaggacct cagtccagag aagtctctag
     3001 ggtctgagga gataaaccca gttcatactg ataatttcca ggagaggggg cctggcaaca
     3061 cagtccaccc ctcagtggct cccatctcct ctgaagagac ccccagccca gagctggtgg
     3121 cctcgggcag ccaggctcct aagatagagg aacccatcca tgccactgca gatccccaga
     3181 aactgagtgt gcttatcact cctgtccccg acacccaggc agccacaagg aggcaggcag
     3241 tggggctact ggctttcctt ggtcttcttt tctgcctagg ggtggccatg tttgcttacc
     3301 agagccttca gggctgtccc cgcaaaatgg cgggggaaat ggtagaaggc ctccgctacg
     3361 tcccccgtag ctgtggcagt aactcatacg tcctggtgcc agtgtgaacc ggtaatcaac
     3421 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     3481 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     3541 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     3601 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     3661 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     3721 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     3781 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
     3841 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     3901 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgagatc
     3961 tgcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     4021 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     4081 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     4141 ggaggattgg gaagacaata gcaggcatgc tggggactcg agttaagggc gaattcccga
     4201 taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact
     4261 acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     4321 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     4381 agcgagcgcg cagcctaatt aaggccttaa ttaaatctgg cgtaatcatg gtcatagctg
     4441 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4501 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4561 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4621 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4681 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4741 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4801 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4861 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4921 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4981 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5041 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5101 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5161 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5221 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5281 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5341 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5401 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5461 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5521 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5581 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5641 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5701 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5761 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5821 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5881 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5941 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6001 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6061 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6121 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6181 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6241 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6301 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6361 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6421 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6481 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6541 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     6601 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     6661 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     6721 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     6781 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     6841 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     6901 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     6961 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     7021 cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     7081 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     7141 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
     7201 taggttaatt aaggctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc
     7261 gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
//
