LOCUS       pAAV_hSyn_NCS3_C        6857 bp DNA     circular SYN 19-APR-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6857)
  AUTHORS   Hochgeschwender
  TITLE     Direct Submission
  JOURNAL   Exported Mar 4, 2024 from SnapGene 7.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6857
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     repeat_region   5..145
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     promoter        190..637
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     primer_bind     419..441
                     /label=D134
     regulatory      657..666
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             663..713
                     /codon_start=1
                     /label=GLuc signal sequence
                     /translation="MGVKVLFALICIAVAEA"
     CDS             join(714..1389,1390..1395,1396..1892)
                     /codon_start=1
                     /label=NCS3
                     /translation="VSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYEE
                     LNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFEDG
                     ASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIST
                     FKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQK
                     AFTGFEDFVGDWEQIAAYNLDQVLEQGGVSSVLQTLAVSVTPIQRIVRSGENGLKIDIH
                     VIIPYEGLSADQMAHIEEVFKVVYPVDDHHFKVIMEYGTLVIDGVTPNMLNYFGRPYEG
                     IAVFDGNKITVTGTLWNGNKIIDERLITPEGSMLFRVTINGVTGYRLLKKISN"
     primer_bind     714..731
                     /label=Lifeact-F
     primer_bind     714..731
                     /label=C564
     primer_bind     1173..1192
                     /label=C644
     primer_bind     complement(1173..1192)
                     /label=C646
     primer_bind     1832..1851
                     /label=C650
     CDS             join(1899..1907,1908..1943)
                     /codon_start=1
                     /label=15aa linker
                     /translation="GDPLVQCGGIAGSAT"
     primer_bind     1899..1917
                     /label=C613
     CDS             1944..2936
                     /label=ChRmine
     CDS             2979..2996
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    3027..3615
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3094..3114)
                     /label=D527
     polyA_signal    3647..4123
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   4163..4303
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4378..4833
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5115..5219
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5220..5288,5289..6080)
                     /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      6251..6839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg
       61 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
      121 gccaactcca tcactagggg ttcctgcggc cgcacgcgtg tgtctagact gcagagggcc
      181 ctgcgtatga gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct
      241 gacgaccgac cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc
      301 cctatcagag agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc
      361 agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac
      421 tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg
      481 tcgcgtccgc gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg
      541 gcacgggcgc gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt
      601 gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg agaaggtacc ggatccgcca
      661 ccatgggagt caaagttctg tttgccctga tctgcatcgc tgtggccgag gccgtgagca
      721 agggcgagga ggataacatg gctagcctcc cagcgacaca tgagttacac atctttggct
      781 ccatcaacgg tgtggacttt gacatggtgg gtcagggcac cggcaatcca aatgatggtt
      841 atgaggagtt aaacctgaag tccaccaagg gtgacctcca gttctccccc tggattctgg
      901 tccctcatat cgggtatggc ttccatcagt acctgcccta ccctgacggg atgtcgcctt
      961 tccaggccgc catggtagat ggctccggat accaagtcca tcgcacaatg cagtttgaag
     1021 atggtgcctc ccttactgtt aactaccgct acacctacga gggaagccac atcaaaggag
     1081 aggcccaggt gaaggggact ggtttccctg ctgacggtcc tgtgatgacc aactcgctga
     1141 ccgctgcgga ctggtgcagg tcgaagaaga cttaccccaa cgacaaaacc atcatcagta
     1201 cctttaagtg gagttacacc actggaaatg gcaagcgcta ccggagcact gcgcggacca
     1261 cctacacctt tgccaagcca atggcggcta actatctgaa gaaccagccg atgtacgtgt
     1321 tccgtaagac ggagctcaag cactccaaga ccgagctcaa cttcaaggag tggcaaaagg
     1381 cctttaccgg atttgaagac ttcgttggtg actgggaaca gatcgctgct tacaacctgg
     1441 accaggttct ggaacagggt ggtgtttctt ctgttctgca gaccctggct gtttctgtta
     1501 cccctatcca gcgtatcgtt cgttctggtg aaaacggtct gaagatcgac atccatgtta
     1561 tcatccctta cgaaggtctg tctgctgacc agatggctca tatcgaagaa gttttcaagg
     1621 ttgtttaccc tgttgacgac catcatttca aggttatcat ggaatacggt accctggtta
     1681 tcgacggtgt tacccctaac atgctgaact acttcggtcg tccttacgaa ggtatcgctg
     1741 ttttcgacgg taataagatc accgttaccg gtaccctgtg gaacggtaac aagatcatcg
     1801 acgaacgtct gatcacccct gaaggttcta tgctgttccg tgttaccatc aacggtgtta
     1861 ccggttaccg tctgctgaag aagatctcta acctcgaggg cgacccacta gtccagtgtg
     1921 gtggaattgc cggatccgcc accgcacacg caccaggcac cgaccagatg ttctacgtgg
     1981 gcacaatgga cggctggtat ctggatacca agctgaactc cgtggccatc ggcgcccact
     2041 ggtcttgctt tatcgtgctg acaatcacca cattctacct gggctatgag tcctggacat
     2101 ctcggggccc tagcaagaga acctcctttt acgccggcta tcaggaggag cagaacctgg
     2161 ccctgtttgt gaatttcttt gccatgctga gctacttcgg caagatcgtg gccgacacac
     2221 tgggccacaa cttcggcgat gtgggcccct tcatcatcgg cttcggcaat tacaggtatg
     2281 ccgactacat gctgacctgc ccaatgctgg tgtacgatct gctgtatcag ctgagggccc
     2341 cctatcgcgt gtcttgtagc gccatcatct ttgccatcct gatgtctggc gtgctggccg
     2401 agttctacgc agagggcgac cctaggctga ggaatggcgc ctacgcctgg tatggctttg
     2461 gctgtttctg gtttatcttc gcctactcta tcgtgatgag catcgtggcc aagcagtata
     2521 gccggctggc ccagctggcc caggatacag gcgccgagca cagcctgcac gtgctgaagt
     2581 tcgccgtgtt taccttctcc atgctgtgga ttctgtttcc cctggtgtgg gccatctgcc
     2641 ctagaggctt cggctggatc gacgataact ggacagaggt ggcccactgc gtgtgcgaca
     2701 tcgtggccaa gtcctgttac ggctttgccc tggcccggtt cagaaagacc tatgatgagg
     2761 agctgtttcg gctgctggag cagctgggac acgacgagga tgagttccag aagctggagc
     2821 tggatatgag gctgagcagc aatggcgagc gcctgcggag actgtctgcg gccgccaaga
     2881 gcaggatcac cagcgagggc gagtacatcc ccctggacca gatcgacatc aacgtgtgaa
     2941 cgcgtggcgc gccaatatta tgctagccaa ttgccggtca tcatcaccat caccattgag
     3001 tttaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     3061 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3121 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     3181 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     3241 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     3301 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     3361 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     3421 aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt
     3481 ccttctgcta cgtcccttcn gccctcaatc cagcgganct tccttcccgc ggcctgctgc
     3541 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gangantcgg atctcccttt
     3601 gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg
     3661 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     3721 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     3781 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     3841 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     3901 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     3961 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     4021 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     4081 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac
     4141 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     4201 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     4261 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     4321 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     4381 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4441 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4501 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4561 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     4621 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4681 tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag
     4741 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4801 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     4861 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     4921 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4981 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     5041 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     5101 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5161 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5221 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5281 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5341 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5401 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5461 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5521 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5581 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5641 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5701 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5761 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5821 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5881 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5941 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6001 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6061 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6121 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6181 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6241 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6301 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6361 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     6421 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6481 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6541 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6601 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6661 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6721 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6781 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6841 acgccagcaa cgcggcc
//
