LOCUS       Exported                6423 bp ds-DNA     circular SYN 25-NOV-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    90 pAAV-ZFN-hSyn-DIO-PSD95.FingR-Citrine-reg.-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6423)
  AUTHORS   Waja Wegner
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Nov 30, 2023 from SnapGene Viewer 3.1.2
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6423
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /note="AAV2 ITR"
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    97..603
                     /note="Sequenziert 13.08.2015"
     misc_feature    102..591
                     /note="seq Endo 15.09.18 fwd"
     misc_feature    156..167
                     /note="ZFN DNA binding domain"
     misc_feature    168..652
                     /note="hSyn promoter"
     misc_feature    654..2574
                     /note="sequenziert 13.08.2015"
     misc_feature    660..1730
                     /note="seq Endo 15.09.18 fwd"
     protein_bind    677..710
                     /bound_moiety="Cre recombinase"
                     /note="lox2272"
                     /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."
     protein_bind    761..794
                     /bound_moiety="Cre recombinase"
                     /note="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(803..2401)
                     /codon_start=1
                     /note="Feature 21"
                     /translation="MLEVKEASPTSIQISWVLHLRHVRYYRITYGETGGNSPVQEFTVP
                     GSKSTATISGLKPGVDYTITVYAVTIFSAYRSAWPPISINYRTGTDYKDDDDKGSGSSR
                     SMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPW
                     PTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEG
                     DTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGS
                     VQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMD
                     ELYKSRGAGAGAGAGAGSRFQCRICMRNFSDRSNLSRHIRTHTGEKPFACDICGRKFAI
                     SSNLNSHTKIHTGSQKPFQCRICMRNFSRSDNLARHIRTHTGEKPFACDICGRKFATSG
                     NLTRHTKIHLRGSQLSIVDAPEQREGASQVSVSVTFEDVAVLFTRDEWKKLDLSQRSLY
                     REVMLENYSNLASMA"
     misc_feature    803..976
                     /note="KRAB-A"
     misc_feature    986..1324
                     /note="CCR5 left-handed ZFN"
     CDS             complement(1370..2086)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /note="Citrine"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    1415..2445
                     /note="seq Endo 15.09.18 fwd"
     CDS             complement(2108..2131)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /note="FLAG"
                     /translation="DYKDDDDK"
     misc_feature    2138..2401
                     /note="PSD95FingR"
     protein_bind    complement(2408..2441)
                     /bound_moiety="Cre recombinase"
                     /note="lox2272"
                     /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."
     protein_bind    complement(2492..2525)
                     /bound_moiety="Cre recombinase"
                     /note="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2550..3138
                     /note="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     polyA_signal    3170..3646
                     /note="hGH poly(A) signal"
                     /note="human growth hormone polyadenylation signal"
     repeat_region   3686..3826
                     /note="AAV2 ITR"
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      3901..4356
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4638..4742
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             4743..5603
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5774..6362
                     /direction=RIGHT
                     /note="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 cgcgtgtcat cctcatcgtg tctagactgc
      181 agagggccct gcgtatgagt gcaagtgggt tttaggacca ggatgaggcg gggtgggggt
      241 gcctacctga cgaccgaccc cgacccactg gacaagcacc caacccccat tccccaaatt
      301 gcgcatcccc tatcagagag ggggagggga aacaggatgc ggcgaggcgc gtgcgcactg
      361 ccagcttcag caccgcggac agtgccttcg cccccgcctg gcggcgcgcg ccaccgccgc
      421 ctcagcactg aaggcgcgct gacgtcactc gccggtcccc cgcaaactcc ccttcccggc
      481 caccttggtc gcgtccgcgc cgccgccggc ccagccggac cgcaccacgc gaggcgcgag
      541 ataggggggc acgggcgcga ccatctgcgc tgcggcgccg gcgactcagc gctgcctcag
      601 tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag cgcagtcgag aaggtaccgg
      661 atccgtcgac tccggaataa cttcgtatag gatactttat acgaagttat gcagaatggt
      721 agctggattg tagctgctat tagcaatatg aaacctctta ataacttcgt atagcataca
      781 ttatacgaag ttatggcgcg ccttaagcca tagaagcaag attagaataa ttttcaagca
      841 taacttcacg ataaagagaa cgttgagaaa gatcaagttt tttccattca tcacgagtaa
      901 aaagaacagc aacatcttca aaagtaacag aaacagaaac ttgagaagca ccttcacgtt
      961 gttcaggagc gtcgacaatc gatagctggc tgccccgcag gtgaatcttt gtgtgccggg
     1021 tcagattgcc ggaggtggca aactttctgc cacagatatc acaggcaaaa ggtttttcac
     1081 cagtatgtgt ccggatgtgt ctggccaggt tgtcgcttct gctaaagttt ctcatacaaa
     1141 ttctgcactg gaaaggtttc tggctgccag tgtggatctt ggtgtggctg ttcaggttgc
     1201 tgctgatggc gaacttccgg ccgcaaatgt cgcaggcgaa gggcttttct ccggtgtggg
     1261 ttctgatgtg cctgctcagg ttggaccggt cgctgaagtt ccgcatgcag atccggcact
     1321 ggaatctaga gccagcccct gcacccgctc cggcgccagc tcccctagac ttgtacagct
     1381 cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc
     1441 gcttctcgtt ggggtctttg ctcagggcgg actggtagct caggtagtgg ttgtcgggca
     1501 gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc
     1561 tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgccgttc ttctgcttgt
     1621 cggccatgat atagacgttg tggctgttgt agttgtactc cagcttgtgc cccaggatgt
     1681 tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct
     1741 cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct
     1801 cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg
     1861 ggtagcgggc gaagcacatc aggccgtagc cgaaggtggt cacgagggtg ggccagggca
     1921 cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag gtggcatcgc
     1981 cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca
     2041 ggatgggcac caccccggtg aacagctcct cgcccttgct caccatggat ctactggagc
     2101 cgctaccctt atcgtcgtca tccttgtaat cggttccggt gcggtagttg atggagatcg
     2161 gcggccaggc ggagcggtag gcgctgaaga tcgtgacggc gtacaccgtg atggtatagt
     2221 cgacaccagg tttcaggccg ctgatggtag cagtggactt gctgccaggc acggtgaatt
     2281 cctggacagg gctattgcca ccagtttcac cgtaggtgat gcggtagtag cgaacgtggc
     2341 gcaagtggag cacccagctg atctggatgc tggttggtga tgcttccttg acttcgagca
     2401 tgctagcata acttcgtata aagtatccta tacgaagtta tttgccttaa cccagaaatt
     2461 atcactgtta ttctttagaa tggtgcaaag aataacttcg tataatgtat gctatacgaa
     2521 gttatgaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     2581 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     2641 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     2701 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     2761 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     2821 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     2881 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     2941 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     3001 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     3061 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     3121 tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc
     3181 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     3241 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     3301 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     3361 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     3421 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     3481 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     3541 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     3601 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt
     3661 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     3721 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     3781 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     3841 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     3901 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3961 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4021 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4081 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     4141 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4201 gactcttgtt ccaaactgga acaacactca accctatctc gggctattct tttgatttat
     4261 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4321 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     4381 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     4441 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4501 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     4561 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4621 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4681 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4741 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4801 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4861 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4921 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4981 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5041 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5101 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5161 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5221 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5281 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5341 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5401 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5461 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5521 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5581 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5641 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5701 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5761 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5821 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5881 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     5941 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6001 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6061 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6121 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6181 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6241 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6301 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6361 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6421 tgt
//
