LOCUS       pAAV-2xcHS4-cjCH        5971 bp    DNA     circular SYN 16-MAY-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5971)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 16, 2025 from SnapGene 8.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5971
                     /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     764..808
                     /label=CHS4_Gib_CHS4_WPRE_BCDel_F
                     /note="color: black; sequence: 
                     catcactaggggttcctgcggccgcacgcgtaCAGGGTAATtgcg; added: 
                     2024-05-23"
     primer_bind     765..817
                     /label=chs4_Gibs_F
                     /note="color: black; sequence: 
                     atcactaggggttcctgcggccgcacgcgtaCAGGGTAATtgcgcgccgagct; 
                     added: 2024-03-14"
     misc_feature    813..1055
                     /locus_tag="cHS4 core(1)"
                     /label=cHS4 core 1
                     /label=cHS4 core(1)
                     /note="color: #00f900"
     misc_feature    1056..1298
                     /locus_tag="cHS4 core"
                     /label=cHS4 core 2
                     /label=cHS4 core
                     /note="color: #00ffff"
     primer_bind     complement(1292..1340)
                     /label=chs4_Gibs_R
                     /note="color: black; sequence: 
                     tctgcagtctagacacttaattaaaCggcgtcggccgtcgatAAGCTTT; added: 
                     2024-03-14"
     primer_bind     1293..1310
                     /label=Primer 2
                     /note="color: black; sequence: AAGCTTatcgatggccga; added: 
                     2022-08-03"
     primer_bind     1333..1383
                     /label=cjCHAT_Ric1_GIbs_F
                     /note="color: black; sequence: 
                     actgcagagggcccgaattcGCATGCAGTCCAGGATGCTTTGCAGGAGTCT; added:
                     2024-09-26"
     primer_bind     complement(1339..1358)
                     /label=CHS4_Gib_CHS4_WPRE_BCDel_R
                     /note="color: black; sequence: 
                     tgattattatagacattgtgtctagaGCATGCgaattcgggccctc; added: 
                     2024-06-01"
     misc_feature    1402..1873
                     /label=cjCHAT_E1
                     /note="chr15_11961481_11961952"
                     /note="color: #008000"
     primer_bind     complement(1906..1954)
                     /label=cjCHAT_Ric1_Gibs_R
                     /note="color: black; sequence: 
                     ctaaagcttatacGATATCACTAGTACCCAATTCCAGGGGACAAGGAAA; added: 
                     2024-09-26"
     primer_bind     1930..1959
                     /label=cjD2full_Gibs_BBswap_middleGFP_F
                     /note="color: black; sequence: 
                     ACTAGTGATATCgtataagctttaggcgtg; added: 2024-09-26"
     misc_feature    1942..2075
                     /label=CMVmini2
                     /note="/vntifkey=21"
                     /note="color: #a6acb3; direction: RIGHT"
     regulatory      2106..2115
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             join(2112..2114,2115..2117,2118..2828)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 2112 .. 2114 / #05fd14
                        2: 2115 .. 2117 / #05fd14 / 1a
                        3: 2118 .. 2828 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2157..2178)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
                     /note="color: black; sequence: CGTCGCCGTCCAGCTCGACCAG"
     primer_bind     complement(2418..2437)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
                     /note="color: black; sequence: GTCTTGTAGTTGCCGTCGTC"
     primer_bind     2765..2786
                     /label=EGFP-C
                     /note="EGFP, forward primer"
                     /note="color: black; sequence: CATGGTCCTGCTGGAGTTCGTG"
     primer_bind     complement(2807..2857)
                     /label=cjD2full_BBswap_middleGFP_R
                     /note="color: black; sequence: 
                     tgtttgaccggtggcgcgccgtcgacttacttgtacagctcgtccatgccg; added:
                     2024-09-26"
     primer_bind     2810..2832
                     /label=Gibson-F-WPRE
                     /note="color: black; sequence: 
                     catggacgagctgtacaagtaaggcgcgccgatgcggggaggcggc; added: 
                     2024-04-27"
     primer_bind     2813..2837
                     /label=GibsonF-WPRE2
                     /note="color: black; sequence: 
                     ggacgagctgtacaagtaagtcgaccgatgcggggaggcggc; added: 
                     2024-04-27"
     misc_feature    2852..2873
                     /label=mmu-miR122-5p Binding site
                     /note="color: #a6acb3"
     misc_feature    2880..2901
                     /label=mmu-miR122-5p Binding site
                     /note="color: #a6acb3"
     misc_feature    2908..2929
                     /label=mmu-miR122-5p Binding site
                     /note="color: #a6acb3"
     misc_feature    2944..2965
                     /label=hsa-miR-183-5p Binding site
                     /note="color: #a6acb3"
     primer_bind     2948..2965
                     /label=WPREFlip_Gib_F
                     /note="color: black; sequence: 
                     aattctaCCAGTGCCATACAAGCTTATCGATCTCGATCGATAATCAACCTCTGGA; 
                     added: 2025-04-29"
     misc_feature    2972..2993
                     /label=hsa-miR-183-5p Binding site
                     /note="color: #a6acb3"
     primer_bind     2976..2993
                     /label=WPREFlip_Gib_F
                     /note="color: black; sequence: 
                     aattctaCCAGTGCCATACAAGCTTATCGATCTCGATCGATAATCAACCTCTGGA; 
                     added: 2025-04-29"
     misc_feature    3000..3021
                     /label=hsa-miR-183-5p Binding site
                     /note="color: #a6acb3"
     primer_bind     3004..3058
                     /label=WPREFlip_Gib_F
                     /note="color: black; sequence: 
                     aattctaCCAGTGCCATACAAGCTTATCGATCTCGATCGATAATCAACCTCTGGA; 
                     added: 2025-04-29"
     primer_bind     3011..3034
                     /label=WPRE_CHS4_miR_F
                     /note="color: black; sequence: 
                     CCAGTGCCATACAAGCTTATCGATGCGGGGAGGCGGCCCA; added: 
                     2024-07-22"
     primer_bind     3034..3059
                     /label=Fragment.REV
                     /note="color: black; sequence: 
                     GCCACCCGTAGATCTCTCGATCGATAATCAACCTCTGGAT; added: 
                     2024-07-22"
     primer_bind     3040..3076
                     /label=Gibson-R-WPRE
                     /note="color: black; sequence: 
                     ggaggggcagagctccaattcgataatcaacctctggattacaaaatttgtgaaag; 
                     added: 2024-04-27"
     misc_feature    3045..3633
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     primer_bind     complement(3098..3118)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
                     /note="color: black; sequence: CATAGCGTAAAAGGAGCAACA; 
                     added: 2025-04-29"
     primer_bind     3583..3633
                     /label=DialOutPrimer1
                     /note="color: black; sequence: 
                     CAAGCAGAAGACGGCATACGAGATTCGCCTTAGTGACTGGAGTTCAGACGTGTGCTCTT
                     CCGATCTCGGATCTCCCTTTGGGCCGCCTCCCCGC; added: 2020-09-15"
     primer_bind     complement(3618..3642)
                     /label=WPRE_CHS4_miR_F
                     /note="color: black; sequence: 
                     CCAGTGCCATACAAGCTTATCGATGCGGGGAGGCGGCCCA; added: 
                     2024-07-22"
     primer_bind     complement(3621..3637)
                     /label=Gibson-F-WPRE
                     /note="color: black; sequence: 
                     catggacgagctgtacaagtaaggcgcgccgatgcggggaggcggc; added: 
                     2024-04-27"
     primer_bind     complement(3621..3637)
                     /label=GibsonF-WPRE2
                     /note="color: black; sequence: 
                     ggacgagctgtacaagtaagtcgaccgatgcggggaggcggc; added: 
                     2024-04-27"
     primer_bind     complement(3623..3671)
                     /label=WPREFlip_Gib_R
                     /note="color: black; sequence: 
                     gaggggtcacagggatGCCACCCGTAGATCTTATCGATGCGGGGAGGCG; added: 
                     2025-04-29"
     primer_bind     complement(3641..3655)
                     /label=Fragment.REV
                     /note="color: black; sequence: 
                     GCCACCCGTAGATCTCTCGATCGATAATCAACCTCTGGAT; added: 
                     2024-07-22"
     polyA_signal    3649..3861
                     /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"
     primer_bind     complement(3876..3900)
                     /label=CHS4_Gib_CHS4_WPRE_BCDel_F
                     /note="color: black; sequence: 
                     catcactaggggttcctgcggccgcacgcgtaCAGGGTAATtgcg; added: 
                     2024-05-23"
     primer_bind     complement(3876..3899)
                     /label=chs4_Gibs_F
                     /note="color: black; sequence: 
                     atcactaggggttcctgcggccgcacgcgtaCAGGGTAATtgcgcgccgagct; 
                     added: 2024-03-14"
     repeat_region   3884..4024
                     /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      4099..4554
                     /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        4836..4940
                     /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             4941..5801
                     /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 gcgtacaggg taattgcgcg ccgagctcac ggggacagcc cccccccaaa
      841 gcccccaggg atgtaattac gtccctcccc cgctaggggg cagcagcgag ccgcccgggg
      901 ctccgctccg gtccggcgct ccccccgcat ccccgagccg gcagcgtgcg gggacagccc
      961 gggcacgggg aaggtggcac gggatcgctt tcctctgaac gcttctcgct gctctttgag
     1021 cctgcagaca cctgggggat acggggaaaa agcttgagct cacggggaca gccccccccc
     1081 aaagccccca gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg
     1141 gggctccgct ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag
     1201 cccgggcacg gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt
     1261 gagcctgcag acacctgggg gatacgggga aaaagcttat cgacggccga cgccgtttaa
     1321 ttaagtgtct agactgcaga gggcccgaat tcgcatgcag tccaggatgc tttgcaggag
     1381 tcttctgact ttcttcaagg aagtacatat cctttctgtc gtagtatgtg ctaaatagaa
     1441 atatcctttg aaaaaaatca ggtctctgag gcaactgtga tgtgaaagtt actagaggaa
     1501 atgttgatac tgatgagcaa atattgccct ttgttaatga gagggactgt acagtctcca
     1561 gacataaaca gctctgtcta gataaaattg tagcagtagg gtgagaataa tgcattgcta
     1621 taattaatgc taatttgtgg atattgctgt attgttgtca ttgtttttta attgaatatg
     1681 tattaagctc taatagggtg ctatgaataa tgccaactta tagttgattt atcagaacac
     1741 tcagaagagg ctttataaac aacaataaat ggaaaatcac agcactcctt cgaggtggta
     1801 attgtctttg ccatttctgc agatgaggag aagtgaagta gattgcattt gctctgttgg
     1861 cactgtgagt gagaacatag aaactgtcaa atgagagaaa agtattttcc ttgtcccctg
     1921 gaattgggta ctagtgatat cgtataagct ttaggcgtgt acggtgggcg cctataaaag
     1981 cagagctcgt ttagtgaacc gtcagatcgc ctggagcaat tccacaacac ttttgtctta
     2041 taccaacttt ccgtaccact tcctaccctc gtaaaggatc cttcgaagct agcagcgctg
     2101 ttaacgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     2161 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     2221 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     2281 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     2341 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     2401 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     2461 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     2521 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     2581 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     2641 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     2701 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     2761 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2821 tgtacaagta agtcgacggc gcgccaccgg tcaaacacca ttgtcacact ccatactatc
     2881 aaacaccatt gtcacactcc agatcaacaa acaccattgt cacactccac gccggcgggt
     2941 accagtgaat tctaccagtg ccatagatca aagtgaattc taccagtgcc atatactata
     3001 gtgaattcta ccagtgccat acaagcttat cgatctcgat cgataatcaa cctctggatt
     3061 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3121 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3181 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     3241 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     3301 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     3361 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     3421 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct
     3481 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     3541 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     3601 cgagtcggat ctccctttgg gccgcctccc cgcatcgata agatctacgg gtggcatccc
     3661 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc
     3721 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
     3781 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
     3841 tgcggggtct attgggaacc acacgtgcgg accgagcggc cgcaggaacc cctagtgatg
     3901 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     3961 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     4021 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4081 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4141 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     4201 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4261 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     4321 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4381 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     4441 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     4501 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     4561 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     4621 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4681 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4741 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     4801 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4861 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4921 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4981 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5041 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5101 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     5161 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5221 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5281 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5341 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5401 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5461 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5521 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5581 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5641 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5701 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5761 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5821 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5881 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5941 cagaccccgt agaaaagatc aaaggatctt c
//
