LOCUS       Exported                6656 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 6656)
  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..6656
                     /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..2354)
                     /codon_start=1
                     /product="modified soybean ascorbate peroxidase (W41F and 
                     A134P), dimeric"
                     /label=dAPEX2
                     /translation="GKSYPTVSADYQKAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAGT
                     FDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVAV
                     EVTGGPEVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGHT
                     IGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVDK
                     YAADEDAFFADYAEAHQKLSELGFADA"
     CDS             complement(2355..2396)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(2406..2609)
                     /codon_start=1
                     /gene="Lactb"
                     /gene_synonym="LACT-1; Lact1; Mrpl56"
                     /product="serine beta-lactamase-like protein LACTB, 
                     mitochondrial precursor"
                     /label=LACTB
                     /note="mitochondrial ribosomal protein L56; serine beta 
                     lactamase-like protein; serine beta-lactamase-like protein 
                     LACTB, mitochondrial"
                     /db_xref="GI:13507666"
                     /db_xref="CCDS:CCDS40674.1"
                     /db_xref="GeneID:80907"
                     /db_xref="MGI:MGI:1933395"
                     /protein_id="NP_109642.1"
                     /translation="MYRLLSSVTARAAATAGPAWDGGRRGAHRRPGLPVLGLGWAGGLG
                     LGLGLALGAKLVVGLRGAVPIQS"
     misc_signal     complement(2610..2615)
                     /label=Kozak consensus sequence (short)
                     /note="vertebrate translation initiation sequence"
     protein_bind    complement(2641..2674)
                     /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(2725..2758)
                     /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    2783..3371
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3254..3265)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3403..3879
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   3919..4059
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4134..4589
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4871..4975
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4976..5836
                     /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      6007..6595
                     /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 aattctaggc atcagcaaac
     1621 ccaagctcgg aaagcttttg gtgagcctca gcgtaatcag caaagaaggc atcttcgtcc
     1681 gctgcatatt tatcaacgag agggcggaat acagggtcag acaaaagagc cttgtcagaa
     1741 ggtagctgaa ggagaccttc cttctcacca ctcaacaact ccgtgaagta tgagttgtcg
     1801 aaaataagag gattagaggt ccagggaccc tcaaatccag aacgctcctt gtgtgcagct
     1861 ccaatagtgt gacccccaga tagagcaacg atatcttggt cagtaagccc catagctttg
     1921 ccaaacacat ctctcaaatg gtcagaaccc ttagtgggat cgggcaagcg accctctggt
     1981 ggtggctcag gcttgtcctc tcttccaggg tggaatggaa cttcaggtcc acccgtgacc
     2041 tcaacggcaa caacgccagc caactggtag aaatcggcgt agctcaaaat agggaactcc
     2101 gccttgagtg gctccaaaag cctaacagcg atgtcaagac cgttgttagc gctgtgagcc
     2161 agttcggcag ggtgcttgat ggttccgaag ggtccaccgg tcttcgtgcc cttgtcaaag
     2221 gttccagcag agtggaatgc caaacggagc attagaggag cgcatctctt ctcagcgatg
     2281 aagcctctga gcttcttctt cgccttctca acggccttct ggtaatcagc actcacagtt
     2341 gggtaagact ttccggtgct gtccaggccc agcagggggt tggggatggg cttgcctgga
     2401 tccttggact gaatggggac ggcgccccgc agcccgacca ccagcttcgc gccgagagcc
     2461 agccccagcc cgagccccag gccgccggcc caaccaaggc ccagcacagg caggcccggt
     2521 cgcctgtgcg ccccgcgccg ccctccgtcc caggctgggc ctgcggtggc cgcagcccga
     2581 gctgtcacgc ttgacaggag ccggtacatg gtggcaattc gtgagcgtcg gttgaacagc
     2641 gaagttccta tactttctag agaataggaa cttcttgcct taacccagaa attatcactg
     2701 ttattcttta gaatggtgca aagagaagtt cctatactat ttgaagaata ggaacttcaa
     2761 tcccgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     2821 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2881 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2941 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3001 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3061 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3121 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     3181 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt
     3241 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3301 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3361 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac
     3421 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     3481 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     3541 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     3601 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     3661 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     3721 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     3781 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     3841 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     3901 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     3961 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     4021 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     4081 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     4141 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4201 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4261 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4321 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     4381 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4441 gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat
     4501 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4561 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     4621 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     4681 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     4741 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     4801 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4861 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4921 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4981 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5041 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5101 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5161 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5221 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5281 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5341 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5401 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5461 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5521 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5581 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5641 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5701 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5761 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5821 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5881 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5941 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6001 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6061 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6121 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     6181 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6241 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6301 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6361 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6421 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6481 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6541 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6601 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
