LOCUS       sequence-324065-        5807 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid expressing soluble CX3CL1 (fractalkine) in retinal 
            pigment epithelium.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5807)
  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 5807)
  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-Best1-sCX3CL1
FEATURES             Location/Qualifiers
     source          1..5807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   46..175
                     /label=AAV2 ITR
     intron          871..1003
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     986..1005
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1023..1032
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2105..2125)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2523..2534)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2601..2618)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2607..2814
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2902..3031
                     /label=AAV2 ITR
     primer_bind     complement(3050..3067)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3050..3066)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3059..3081)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3208..3663
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3295..3314)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3505..3526
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3689..3793
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3794..3862,3863..4654)
                     /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(4012..4031)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4825..5413
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5314..5333
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5701..5722
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5737..5742,5743..5760,5761..5767)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5775..5791
                     /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     5780..5802
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(5799..5807,1..8)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5799..5807,1..8)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gctatgacca tgattacgcc agatttaatt aaggccttaa ttaggctgcg cgctcgctcg
       61 ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca
      121 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttccttgtag
      181 ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa gatcggaatt
      241 cgcccttaag ctagctcctt ttcagataag ggcactgagg ctgagagagg agctgaaacc
      301 tacccggggt caccacacac aggtggcaag gctgggacca gaaaccagga ctgttgactg
      361 cagcccggta ttcattcttt ccatagccca cagggctgtc aaagacccca gggcctagtc
      421 agaggctcct ccttcctgga gagttcctgg cacagaagtt gaagctcagc acagccccct
      481 aacccccaac tctctctgca aggcctcagg ggtcagaaca ctggtggagc agatccttta
      541 gcctctggat tttagggcca tggtagaggg ggtgttgccc taaattccag ccctggtctc
      601 agcccaacac cctccaagaa gaaattagag gggccatggc caggctgtgc tagccgttgc
      661 ttctgagcag attacaagaa gggactaaga caaggactcc tttgtggagg tcctggctta
      721 gggagtcaag tgacggcggc tcagcactca cgtgggcagt gccagcctct aagagtgggc
      781 aggggcactg gccacagagt cccagggagt cccaccagcc tagtcgccag acccagatcc
      841 tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa
      901 ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca
      961 cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca ggcggccgcg
     1021 ccgccaccat ggctccctcg ccgctcgcgt ggctgctgcg cctggccgcg ttcttccatt
     1081 tgtgtactct gctgccgggt cagcacctcg gcatgacgaa atgcgaaatc atgtgcgaca
     1141 agatgacctc acgaatccca gtggctttgc tcatccgcta tcagctaaac caggagtcct
     1201 gcggcaagcg tgccattgtc ctggagacga cacagcacag acgcttctgt gctgacccga
     1261 aggagaaatg ggtccaagac gccatgaagc atctggatca ccaggctgct gccctcacta
     1321 aaaatggtgg caagtttgag aagcgggtgg acaatgtgac acctgggatc accttggcca
     1381 ctaggggact gtccccatct gccctgacaa agcctgaatc cgccacattg gaagaccttg
     1441 ctttggaact gactactatt tcccaggagg ccagggggac catggggact tcccaagagc
     1501 caccggcagc agtgaccgga tcatctctct caacttccga ggcacaggat gcagggctta
     1561 cggctaagcc tcagagcatt ggaagttttg aggcggctga catctccacc accgtttggc
     1621 cgagtcctgc tgtctaccaa tctggatcta gctcctgggc tgaggaaaaa gctactgagt
     1681 ccccctccac tacagcccca tctcctcagg tgtccactac ttcaccttca accccagagg
     1741 aaaatgttgg gtccgaaggc caacccccat gggtccaggg acaggacctc agtccagaga
     1801 agtctctagg gtctgaggag ataaacccag ttcatactga taatttccag gagagggggc
     1861 ctggcaacac agtccacccc tcagtggctc ccatctcctc tgaagagacc cccagcccag
     1921 agctggtggc ctcgggcagc caggctccta agatagagga acccatccat gccactgcag
     1981 atccccagaa actgagtgtg cttatcactc ctgtccccga cacccaggca gccacatgaa
     2041 agcttggatc caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     2101 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     2161 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     2221 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     2281 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     2341 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     2401 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     2461 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     2521 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     2581 ttccgcgtct tcgagatctg cctcgactgt gccttctagt tgccagccat ctgttgtttg
     2641 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2701 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     2761 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggactcgag
     2821 ttaagggcga attcccgata aggatcttcc tagagcatgg ctacgtagat aagtagcatg
     2881 gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc
     2941 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     3001 gggcggcctc agtgagcgag cgagcgcgca gccttaatta acctaattca ctggccgtcg
     3061 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     3121 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     3181 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     3241 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3301 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3361 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3421 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3481 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3541 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3601 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     3661 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3721 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3781 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3841 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3901 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3961 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4021 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4081 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4141 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4201 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4261 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4321 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4381 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4441 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4501 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4561 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4621 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4681 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4741 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4801 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4861 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4921 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4981 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5041 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5101 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5161 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5221 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5281 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5341 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5401 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5461 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5521 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     5581 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     5641 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     5701 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     5761 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaaca
//
