LOCUS       Exported                6485 bp ds-DNA     circular SYN 28-JUN-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    PT-2788   rAAV-CMV-DIO-(mCherry-U6)-shRNA(scramble)-WPRE-hGH p...
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6485)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 14, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6485
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    221..508
                     /label=CAG Enhancer
     promoter        521..723
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        816..1308
                     /label=beta-globin intron
     misc_feature    1193..1214
                     /label=CMV-1153-C-F
     misc_feature    1315..1341
                     /label=F
     protein_bind    1327..1360
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     misc_feature    1399..1437
                     /label=R
     protein_bind    1411..1444
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    1445..1473
                     /label=F
     promoter        complement(1449..1689)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    1665..1689
                     /label=R
     CDS             complement(1696..2406)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEYNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     protein_bind    complement(2419..2452)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     misc_feature    2494..2532
                     /label=R
     protein_bind    complement(2503..2536)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2542..2594
                     /label=shRNA(scramble)
     misc_feature    2612..3200
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     misc_feature    2665..2685
                     /label=WPRE-R  擎科通用测序引物  CATAGCGTAAAAGGAGCAACA
     polyA_signal    3232..3708
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     misc_feature    3403..3423
                     /label=hGH polyA-N-R   ggccctacaggttgtcttccc
     repeat_region   3748..3888
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      3963..4418
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4700..4804
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4805..5665
                     /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"
     rep_origin      5836..6424
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtctagt tattaatagt aatcaattac
      181 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      241 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      301 catagtaacg tcaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      361 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      421 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      481 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      541 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      601 cgtcaatggg agtttgtttt gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
      661 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
      721 gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat
      781 agaagacacc gggaccgatc cagcctccgc ggattcgaat cccggccggg aacggtgcat
      841 tggaacgcgg attccccgtg ccaagagtga cgtaagtacc gcctatagag tctataggcc
      901 cacaaaaaat gctttcttct tttaatatac ttttttgttt atcttatttc taatactttc
      961 cctaatctct ttctttcagg gcaataatga tacaatgtat catgcctctt tgcaccattc
     1021 taaagaataa cagtgataat ttctgggtta aggcaatagc aatatttctg catataaata
     1081 tttctgcata taaattgtaa ctgatgtaag aggtttcata ttgctaatag cagctacaat
     1141 ccagctacca ttctgctttt attttatggt tgggataagg ctggattatt ctgagtccaa
     1201 gctaggccct tttgctaatc atgttcatac ctcttatctt cctcccacag ctcctgggca
     1261 acgtgctggt ctgtgtgctg gcccatcact ttggcaaaga attgggattc tagagtcgac
     1321 tccggaataa cttcgtatag gatactttat acgaagttat gcagaatggt agctggattg
     1381 tagctgctat tagcaatatg aaacctctta ataacttcgt atagcataca ttatacgaag
     1441 ttattttcgt cctttccaca agatatataa agccaagaaa tcgaaatact ttcaagttac
     1501 ggtaagcata tgatagtcca ttttaaaaca taattttaaa actgcaaact acccaagaaa
     1561 ttattacttt ctacgtcacg tattttgtac taatatcttt gtgtttacag tcaaattaat
     1621 tccaattatc tctctaacag ccttgtatcg tatatgcaaa tatgaaggaa tcatgggaaa
     1681 taggccctcg gatccttact tgtacagctc gtccatgccg ccggtggagt ggcggccctc
     1741 ggcgcgttcg tactgttcca cgatggtgta gtcctcgttg tgggaggtga tgtccaactt
     1801 gatgttgacg ttgtaggcgc cgggcagctg cacgggcttc ttggccttgt aggtggtctt
     1861 gacctcagcg tcgtagtggc cgccgtcctt cagcttcagc ctctgcttga tctcgccctt
     1921 cagggcgccg tcctcggggt acatccgctc ggaggaggcc tcccagccca tggtcttctt
     1981 ctgcattacg gggccgtcgg aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa
     2041 ctcgccgtcc tgcagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt
     2101 catcacgcgc tcccacttga agccctcggg gaaggacagc ttcaagtagt cggggatgtc
     2161 ggcggggtgc ttcacgtagg ccttggagcc gtacatgaac tgaggggaca ggatgtccca
     2221 ggcgaagggc agggggccac ccttggtcac cttcagcttg gcggtctggg tgccctcgta
     2281 ggggcggccc tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat
     2341 gtgcaccttg aagcgcatga actccttgat gatggccatg ttatactcct cgcccttgct
     2401 caccatggtg gcgctagcat aacttcgtat aaagtatcct atacgaagtt atttgcctta
     2461 acccagaaat tatcactgtt attctttaga atggtgcaaa gaataacttc gtataatgta
     2521 tgctatacga agttatcacc ggcctaaggt taagtcgccc tcgttcaaga gacgagggcg
     2581 acttaacctt aggctttttt aattcactag taatcaacct ctggattaca aaatttgtga
     2641 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2701 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2761 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2821 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2881 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2941 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     3001 ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg
     3061 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3121 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3181 cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat
     3241 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     3301 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     3361 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     3421 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     3481 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc
     3541 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     3601 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc
     3661 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag
     3721 gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     3781 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     3841 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     3901 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     3961 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4021 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4081 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4141 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     4201 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4261 tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt
     4321 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4381 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     4441 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     4501 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4561 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     4621 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     4681 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4741 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4801 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4861 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4921 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4981 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5041 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5101 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5161 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5221 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5281 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5341 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5401 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5461 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5521 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5581 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5641 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5701 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5761 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5821 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5881 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5941 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     6001 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6061 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6121 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6181 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6241 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6301 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6361 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6421 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6481 catgt
//
