LOCUS       Exported                6476 bp ds-DNA     circular SYN 06-SEP-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6476)
  AUTHORS   HMS Neurobiology
  TITLE     Direct Submission
  JOURNAL   Exported Sep 6, 2018 from SnapGene 4.2.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6476
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     promoter        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     protein_bind    1450..1483
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    1534..1567
                     /label=FRT3 (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="minimal FRT site with mutations in the spacer 
                     sequence, supports FLP-mediated excision but not 
                     integration, incompatible with the WT FRT site (Schlake and
                     Bode, 1994)"
     CDS             complement(1608..1619)
                     /codon_start=1
                     /label=KDEL
                     /translation="KDEL"
     CDS             complement(1620..2366)
                     /codon_start=1
                     /product="modified soybean ascorbate peroxidase (W41F and 
                     A134P), dimeric"
                     /label=dAPEX2
                     /translation="GKSYPTVSADYQKAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAGT
                     FDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVAV
                     EVTGGPEVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGHT
                     IGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVDK
                     YAADEDAFFADYAEAHQKLSELGFADA"
     CDS             complement(2367..2429)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     misc_signal     complement(2426..2435)
                     /label=Kozak consensus sequence
                     /note="vertebrate translation initiation sequence"
     protein_bind    complement(2461..2494)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    complement(2545..2578)
                     /label=FRT3 (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="minimal FRT site with mutations in the spacer 
                     sequence, supports FLP-mediated excision but not 
                     integration, incompatible with the WT FRT site (Schlake and
                     Bode, 1994)"
     misc_feature    2603..3191
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3074..3085)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3223..3699
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   3739..3879
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      3954..4409
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4691..4795
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4796..5656
                     /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      5827..6415
                     /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 cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc
     1441 gactccggag aagttcctat tctctagaaa gtataggaac ttcgcagaat ggtagctgga
     1501 ttgtagctgc tattagcaat atgaaacctc ttggaagttc ctattcttca aatagtatag
     1561 gaacttcgcc tcaaagccgc gtgacgagct agcccatggg aattttacag ctcgtccttg
     1621 gcatcagcaa acccaagctc ggaaagcttt tggtgagcct cagcgtaatc agcaaagaag
     1681 gcatcttcgt ccgctgcata tttatcaacg agagggcgga atacagggtc agacaaaaga
     1741 gccttgtcag aaggtagctg aaggagacct tccttctcac cactcaacaa ctccgtgaag
     1801 tatgagttgt cgaaaataag aggattagag gtccagggac cctcaaatcc agaacgctcc
     1861 ttgtgtgcag ctccaatagt gtgaccccca gatagagcaa cgatatcttg gtcagtaagc
     1921 cccatagctt tgccaaacac atctctcaaa tggtcagaac ccttagtggg atcgggcaag
     1981 cgaccctctg gtggtggctc aggcttgtcc tctcttccag ggtggaatgg aacttcaggt
     2041 ccacccgtga cctcaacggc aacaacgcca gccaactggt agaaatcggc gtagctcaaa
     2101 atagggaact ccgccttgag tggctccaaa agcctaacag cgatgtcaag accgttgtta
     2161 gcgctgtgag ccagttcggc agggtgcttg atggttccga agggtccacc ggtcttcgtg
     2221 cccttgtcaa aggttccagc agagtggaat gccaaacgga gcattagagg agcgcatctc
     2281 ttctcagcga tgaagcctct gagcttcttc ttcgccttct caacggcctt ctggtaatca
     2341 gcactcacag ttgggtaaga ctttccgtca ccagtggaac ctggaaccca gagcagcagt
     2401 acccatagca ggagtgtgtc tgtctccatg gtggcaattc gtgagcgtcg gttgaacagc
     2461 gaagttccta tactttctag agaataggaa cttcttgcct taacccagaa attatcactg
     2521 ttattcttta gaatggtgca aagagaagtt cctatactat ttgaagaata ggaacttcaa
     2581 tcccgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     2641 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2701 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2761 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2821 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2881 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     2941 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     3001 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt
     3061 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3121 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3181 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac
     3241 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     3301 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     3361 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     3421 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     3481 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     3541 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     3601 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     3661 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     3721 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     3781 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     3841 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     3901 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     3961 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4021 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4081 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4141 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     4201 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4261 gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat
     4321 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4381 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     4441 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     4501 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     4561 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     4621 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4681 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4741 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4801 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4861 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4921 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4981 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5041 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5101 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5161 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5221 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5281 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5341 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5401 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5461 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5521 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5581 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5641 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5701 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5761 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5821 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5881 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5941 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     6001 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6061 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6121 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6181 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6241 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6301 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6361 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6421 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
