LOCUS       pAAV-2xCHS4-cjBC        5989 bp    DNA     circular SYN 16-MAY-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5989)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 16, 2025 from SnapGene 8.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5989
                     /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     complement(1339..1359)
                     /label=CHS4_Gib_CHS4_WPRE_BCDel_R
                     /note="color: black; sequence: 
                     tgattattatagacattgtgtctagaGCATGCgaattcgggccctc; added: 
                     2024-06-01"
     misc_feature    1359..1947
                     /label=ePFC_BCL11B_4
                     /note="color: #339966"
     primer_bind     1948..1977
                     /label=cjD2full_Gibs_BBswap_middleGFP_F
                     /note="color: black; sequence: 
                     ACTAGTGATATCgtataagctttaggcgtg; added: 2024-10-17"
     misc_feature    1960..2093
                     /label=CMVmini2
                     /note="/vntifkey=21"
                     /note="color: #a6acb3; direction: RIGHT"
     regulatory      2124..2133
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             join(2130..2132,2133..2135,2136..2846)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="This feature has 3 segments:
                        1: 2130 .. 2132 / #05fc14
                        2: 2133 .. 2135 / #05fc14 / 1a
                        3: 2136 .. 2846 / #05fc14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2175..2196)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
                     /note="color: black; sequence: CGTCGCCGTCCAGCTCGACCAG"
     primer_bind     complement(2436..2455)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
                     /note="color: black; sequence: GTCTTGTAGTTGCCGTCGTC"
     primer_bind     2783..2804
                     /label=EGFP-C
                     /note="EGFP, forward primer"
                     /note="color: black; sequence: CATGGTCCTGCTGGAGTTCGTG"
     primer_bind     complement(2825..2875)
                     /label=cjD2full_BBswap_middleGFP_R
                     /note="color: black; sequence: 
                     tgtttgaccggtggcgcgccgtcgacttacttgtacagctcgtccatgccg; added:
                     2024-10-17"
     primer_bind     2828..2850
                     /label=Gibson-F-WPRE
                     /note="color: black; sequence: 
                     catggacgagctgtacaagtaaggcgcgccgatgcggggaggcggc; added: 
                     2024-04-27"
     primer_bind     2831..2855
                     /label=GibsonF-WPRE2
                     /note="color: black; sequence: 
                     ggacgagctgtacaagtaagtcgaccgatgcggggaggcggc; added: 
                     2024-04-27"
     misc_feature    2870..2891
                     /label=mmu-miR122-5p Binding site
                     /note="color: #a6acb3"
     misc_feature    2898..2919
                     /label=mmu-miR122-5p Binding site
                     /note="color: #a6acb3"
     misc_feature    2926..2947
                     /label=mmu-miR122-5p Binding site
                     /note="color: #a6acb3"
     misc_feature    2962..2983
                     /label=hsa-miR-183-5p Binding site
                     /note="color: #a6acb3"
     primer_bind     2966..2983
                     /label=WPREFlip_Gib_F
                     /note="color: black; sequence: 
                     aattctaCCAGTGCCATACAAGCTTATCGATCTCGATCGATAATCAACCTCTGGA; 
                     added: 2025-04-30"
     misc_feature    2990..3011
                     /label=hsa-miR-183-5p Binding site
                     /note="color: #a6acb3"
     primer_bind     2994..3011
                     /label=WPREFlip_Gib_F
                     /note="color: black; sequence: 
                     aattctaCCAGTGCCATACAAGCTTATCGATCTCGATCGATAATCAACCTCTGGA; 
                     added: 2025-04-30"
     misc_feature    3018..3039
                     /label=hsa-miR-183-5p Binding site
                     /note="color: #a6acb3"
     primer_bind     3022..3076
                     /label=WPREFlip_Gib_F
                     /note="color: black; sequence: 
                     aattctaCCAGTGCCATACAAGCTTATCGATCTCGATCGATAATCAACCTCTGGA; 
                     added: 2025-04-30"
     primer_bind     3029..3052
                     /label=WPRE_CHS4_miR_F
                     /note="color: black; sequence: 
                     CCAGTGCCATACAAGCTTATCGATGCGGGGAGGCGGCCCA; added: 
                     2024-07-22"
     primer_bind     3052..3077
                     /label=Fragment.REV
                     /note="color: black; sequence: 
                     GCCACCCGTAGATCTCTCGATCGATAATCAACCTCTGGAT; added: 
                     2024-07-22"
     primer_bind     3058..3094
                     /label=Gibson-R-WPRE
                     /note="color: black; sequence: 
                     ggaggggcagagctccaattcgataatcaacctctggattacaaaatttgtgaaag; 
                     added: 2024-04-27"
     misc_feature    3063..3651
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     primer_bind     complement(3116..3136)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
                     /note="color: black; sequence: CATAGCGTAAAAGGAGCAACA"
     primer_bind     3601..3651
                     /label=DialOutPrimer1
                     /note="color: black; sequence: 
                     CAAGCAGAAGACGGCATACGAGATTCGCCTTAGTGACTGGAGTTCAGACGTGTGCTCTT
                     CCGATCTCGGATCTCCCTTTGGGCCGCCTCCCCGC; added: 2020-09-15"
     primer_bind     complement(3636..3660)
                     /label=WPRE_CHS4_miR_F
                     /note="color: black; sequence: 
                     CCAGTGCCATACAAGCTTATCGATGCGGGGAGGCGGCCCA; added: 
                     2024-07-22"
     primer_bind     complement(3639..3655)
                     /label=Gibson-F-WPRE
                     /note="color: black; sequence: 
                     catggacgagctgtacaagtaaggcgcgccgatgcggggaggcggc; added: 
                     2024-04-27"
     primer_bind     complement(3639..3655)
                     /label=GibsonF-WPRE2
                     /note="color: black; sequence: 
                     ggacgagctgtacaagtaagtcgaccgatgcggggaggcggc; added: 
                     2024-04-27"
     primer_bind     complement(3641..3689)
                     /label=WPREFlip_Gib_R
                     /note="color: black; sequence: 
                     gaggggtcacagggatGCCACCCGTAGATCTTATCGATGCGGGGAGGCG; added: 
                     2025-04-30"
     primer_bind     complement(3659..3673)
                     /label=Fragment.REV
                     /note="color: black; sequence: 
                     GCCACCCGTAGATCTCTCGATCGATAATCAACCTCTGGAT; added: 
                     2024-07-22"
     polyA_signal    3667..3879
                     /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(3894..3918)
                     /label=CHS4_Gib_CHS4_WPRE_BCDel_F
                     /note="color: black; sequence: 
                     catcactaggggttcctgcggccgcacgcgtaCAGGGTAATtgcg; added: 
                     2024-05-23"
     primer_bind     complement(3894..3917)
                     /label=chs4_Gibs_F
                     /note="color: black; sequence: 
                     atcactaggggttcctgcggccgcacgcgtaCAGGGTAATtgcgcgccgagct; 
                     added: 2024-03-14"
     repeat_region   3902..4042
                     /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      4117..4572
                     /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        4854..4958
                     /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             4959..5819
                     /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 tcgcatgctg gaccctcccc agcaaggaaa
     1381 gagacacggg tgtcaggagc tctgaagtct gtgcaaccct ataaatctac ctgtgttcca
     1441 caatgttgag ccccttatat gtgtgcactg aggaacggct gcattggcac aggagtccta
     1501 gactcccaga ctttgtgctc caccctgaaa tagcatccag tttcatggca ggctctgttt
     1561 ttttccaagg aagttaaata tgcacagccc gcactttgca ataaaatcaa ggttatgcaa
     1621 aaaagtaatt cagctctggc ttccaggaat ccccgtggaa aatatatatg tgtctattta
     1681 taaagaaaaa attaatacag caaacaatct cattgaagga aacttctaaa tgtgttccaa
     1741 caaaacctgt cccggagtca gcaactcctc ccgcctccca cttcagagga gcagagactt
     1801 caagttctgg tttaaattta gatgctgtct tttctgatgg gaatttgaaa gttgggggaa
     1861 aatatgcacc aagtatatat taggattagg aaacatccgt agaatctatt tcctgaactt
     1921 cataaacaga ggaactaaat tatcactact agtgatatcg tataagcttt aggcgtgtac
     1981 ggtgggcgcc tataaaagca gagctcgttt agtgaaccgt cagatcgcct ggagcaattc
     2041 cacaacactt ttgtcttata ccaactttcc gtaccacttc ctaccctcgt aaaggatcct
     2101 tcgaagctag cagcgctgtt aacgccacca tggtgagcaa gggcgaggag ctgttcaccg
     2161 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     2221 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     2281 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     2341 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     2401 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     2461 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     2521 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     2581 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     2641 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     2701 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     2761 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     2821 ctctcggcat ggacgagctg tacaagtaag tcgacggcgc gccaccggtc aaacaccatt
     2881 gtcacactcc atactatcaa acaccattgt cacactccag atcaacaaac accattgtca
     2941 cactccacgc cggcgggtac cagtgaattc taccagtgcc atagatcaaa gtgaattcta
     3001 ccagtgccat atactatagt gaattctacc agtgccatac aagcttatcg atctcgatcg
     3061 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     3121 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     3181 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     3241 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     3301 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     3361 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     3421 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     3481 tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     3541 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     3601 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgataag
     3661 atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc
     3721 actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag
     3781 gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg
     3841 aagacaacct gtagggcctg cggggtctat tgggaaccac acgtgcggac cgagcggccg
     3901 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     3961 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     4021 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     4081 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     4141 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     4201 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4261 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4321 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4381 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4441 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     4501 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     4561 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4621 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4681 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4741 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     4801 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4861 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4921 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4981 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5041 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5101 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5161 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5221 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5281 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5341 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5401 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5461 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5521 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5581 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5641 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5701 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5761 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5821 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5881 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5941 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttc
//
