LOCUS       pAAV-2xcHS4-cjD2        5451 bp    DNA     circular SYN 16-MAY-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5451)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported May 16, 2025 from SnapGene 8.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5451
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     rep_origin      1..589
                     /label=ori
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=rep_origin"
                     /note="color: #ffff00"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=repeat region"
                     /note="color: #a6acb3"
     primer_bind     811..866
                     /label=cjD2full_Gibs_BBswap_R
                     /note="color: black; sequence: 
                     actgcagagggcccgaattcGCATGCTACAAATGAAGCGTAAATCTACCCAGTGAG; 
                     added: 2024-02-15"
     misc_feature    836..1754
                     /label=cjD2-E8 enhancer
                     /note="color: #339966"
     primer_bind     complement(1224..1249)
                     /label=cj_D2enh_small_right_F
                     /note="color: black; sequence: 
                     gtacgCTCGAGGGTATGTGTGCAATGGAGATGGGTTG; added: 2023-04-05"
     primer_bind     1250..1280
                     /label=cj_D2enh_small_left_R
                     /note="color: black; sequence: 
                     agctgTCTAGAATGCTCAAAGTCTGTGAAAGATGTTGACCTG; added: 
                     2023-04-05"
     primer_bind     complement(1734..1779)
                     /label=cjD2full_Gibs_BBswap_F
                     /note="color: black; sequence: 
                     ctaaagcttatacGATATCACTAGTGAGGCTTCCTTTCTGCCCTGC; added: 
                     2024-02-15"
     primer_bind     1755..1784
                     /label=cjD2full_Gibs_BBswap_middleGFP_F
                     /note="color: black; sequence: 
                     ACTAGTGATATCgtataagctttaggcgtg; added: 2024-02-15"
     misc_feature    1767..1900
                     /label=CMVmini2
                     /note="/vntifkey=21"
                     /note="color: #a6acb3; direction: RIGHT"
     regulatory      1931..1940
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             join(1937..1939,1940..1942,1943..2653)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="This feature has 3 segments:
                        1: 1937 .. 1939 / #05fc14
                        2: 1940 .. 1942 / #05fc14 / 1a
                        3: 1943 .. 2653 / #05fc14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1982..2003)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
                     /note="color: black; sequence: CGTCGCCGTCCAGCTCGACCAG"
     primer_bind     complement(2243..2262)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
                     /note="color: black; sequence: GTCTTGTAGTTGCCGTCGTC"
     primer_bind     2590..2611
                     /label=EGFP-C
                     /note="EGFP, forward primer"
                     /note="color: black; sequence: CATGGTCCTGCTGGAGTTCGTG"
     primer_bind     complement(2632..2682)
                     /label=cjD2full_BBswap_middleGFP_R
                     /note="color: black; sequence: 
                     tgtttgaccggtggcgcgccgtcgacttacttgtacagctcgtccatgccg; added:
                     2024-02-15"
     misc_feature    2677..2698
                     /label=mmu-miR122-5p Binding site
                     /note="color: #a6acb3"
     misc_feature    2705..2726
                     /label=mmu-miR122-5p Binding site
                     /note="color: #a6acb3"
     misc_feature    2733..2754
                     /label=mmu-miR122-5p Binding site
                     /note="color: #a6acb3"
     misc_feature    2769..2790
                     /label=hsa-miR-183-5p Binding site
                     /note="color: #a6acb3"
     misc_feature    2797..2818
                     /label=hsa-miR-183-5p Binding site
                     /note="color: #a6acb3"
     misc_feature    2825..2846
                     /label=hsa-miR-183-5p Binding site
                     /note="color: #a6acb3"
     misc_feature    2857..3107
                     /label=WPRE3
                     /note="color: #99cc00"
     polyA_signal    3129..3341
                     /label=hGH poly(A) signal
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=True"
                     /note="/identity=97.685"
                     /note="/match_length=45.093945720250524"
                     /note="/other=polyA signal"
                     /note="color: #a6acb3"
     repeat_region   3364..3504
                     /label=AAV2 ITR
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=repeat region"
                     /note="color: #a6acb3"
     rep_origin      3579..4034
                     /label=f1 ori
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=rep_origin"
                     /note="color: #ffff00"
     promoter        4316..4420
                     /label=AmpR promoter
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
                     /note="color: #ffffff"
     CDS             4421..5281
                     /codon_start=1
                     /label=AmpR
                     /note="pLannotate"
                     /note="/database=addgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /note="color: #993366"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gcgtttaatt aagtgtctag actgcagagg gcccgaattc gcatgctaca
      841 aatgaagcgt aaatctaccc agtgagggag gaatattctt accaccttga gaataaccgc
      901 ggtgatgaca aacaaggggc tggcacccac gggctcaggc tctggggagc tgtcagggcg
      961 taatttgcat gctaaataca gtatttttag caccaaagtt tggagcactt atcttgccct
     1021 gaacagttaa ttatggactt ttgtcttctc ccgaacctaa aggtagtgct gagccctggg
     1081 agaggctcct ctttccaaga gcaccctgat tctggaaggt gggcaaaaca ggcccctagt
     1141 ctaacttagg gggaccactg gcccagtgac taccttctgg ggaaggcatc aggtccccaa
     1201 gagggataga agccccagga acccaaccca tctccattgc acacatacca tgctcaaagt
     1261 ctgtgaaaga tgttgacctg ggtggacgct taaaagcata acccacgatt aggaatctta
     1321 cgagggtata caggcttatc agctgtgagg attggaagag aaaacaaagg gaaacagagg
     1381 aaagaagaag gaagggaggg agagagagag ggagtgaagc aagacccaga gcagagtgag
     1441 aagagcattg acagggagca gaggggaaga gggcagggca ggggcgggag gcggggcagg
     1501 ggaaagttgc ccacagttgt cacgaggctt cacgtctttc taccagacag cagattgaca
     1561 gctagagtgg gcagggggag ggttgggatc caccctctcc cccctcagca cttcaggggc
     1621 aaagaaatgg gaggagtagg acctgacacg acacgggaac acaaggtgga aggggtggcc
     1681 caggctctga ctctctccag agaggccctg acgatggtct cttgctcttg acggcagggc
     1741 agaaaggaag cctcactagt gatatcgtat aagctttagg cgtgtacggt gggcgcctat
     1801 aaaagcagag ctcgtttagt gaaccgtcag atcgcctgga gcaattccac aacacttttg
     1861 tcttatacca actttccgta ccacttccta ccctcgtaaa ggatccttcg aagctagcag
     1921 cgctgttaac gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     1981 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     2041 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     2101 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     2161 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     2221 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     2281 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     2341 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     2401 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     2461 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     2521 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     2581 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     2641 cgagctgtac aagtaagtcg acggcgcgcc accggtcaaa caccattgtc acactccata
     2701 ctatcaaaca ccattgtcac actccagatc aacaaacacc attgtcacac tccacgccgg
     2761 cgggtaccag tgaattctac cagtgccata gatcaaagtg aattctacca gtgccatata
     2821 ctatagtgaa ttctaccagt gccatacaag cttatcgata atcaacctct ggattacaaa
     2881 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     2941 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3001 ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg ccttgcccgc
     3061 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttagc gctgctcgag
     3121 agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     3181 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     3241 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     3301 ggaagacaac ctgtagggcc tgcggggtct attgggaacc acacgtgcgg accgagcggc
     3361 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     3421 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     3481 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     3541 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     3601 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     3661 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3721 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3781 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3841 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3901 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     3961 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     4021 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     4081 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     4141 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     4201 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     4261 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     4321 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4381 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4441 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4501 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4561 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4621 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4681 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4741 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4801 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4861 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4921 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4981 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5041 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5101 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5161 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5221 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5281 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5341 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5401 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt c
//
