LOCUS       Exported                6789 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV-mediated expression of Chronos-tdTomato under the Syn promoter. 
            Using bGHpA signal. tdTomato has codons varied between the first and
            second tandem repeats to reduce recombination..
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-Syn-Chronos-tdTomato
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6789)
  AUTHORS   Klapoetke NC, Murata Y, Kim SS, Pulver SR, Birdsey-Benson A, Cho YK,
            Morimoto TK, Chuong AS, Carpenter EJ, Tian Z, Wang J, Xie Y, Yan Z, 
            Zhang Y, Chow BY, Surek B, Melkonian M, Jayaraman V, 
            Constantine-Paton M, Wong GK, Boyden ES
  TITLE     Independent optical excitation of distinct neural populations.
  JOURNAL   Nat Methods. 2014 Mar;11(3):338-46. doi: 10.1038/nmeth.2836. Epub 
            2014 Feb 9.
  PUBMED    24509633
REFERENCE   2  (bases 1 to 6789)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6789
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   46..175
                     /label=AAV2 ITR
     promoter        275..722
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      735..744
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    3186..3774
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3239..3259)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3657..3668)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3815..4022
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4192..4321
                     /label=AAV2 ITR
     primer_bind     complement(4387..4404)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4558..5146)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4638..4657)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5317..6177)
                     /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"
     primer_bind     5940..5959
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6178..6282)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6308..6763)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6445..6466)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6657..6676
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctgcg cgctcgctcg
       61 ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca
      121 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttccttgtag
      181 ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa gatcgtaggt
      241 taattaatct agactgcaga gggccctgcg tatgagtgca agtgggtttt aggaccagga
      301 tgaggcgggg tgggggtgcc tacctgacga ccgaccccga cccactggac aagcacccaa
      361 cccccattcc ccaaattgcg catcccctat cagagagggg gaggggaaac aggatgcggc
      421 gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt gccttcgccc ccgcctggcg
      481 gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc ggtcccccgc
      541 aaactcccct tcccggccac cttggtcgcg tccgcgccgc cgccggccca gccggaccgc
      601 accacgcgag gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc ggcgccggcg
      661 actcagcgct gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc ctgagagcgc
      721 agtcgagagg atccgccacc atggaaacag ccgccacaat gacccacgcc tttatctcag
      781 ccgtgcctag cgccgaagcc acaattagag gcctgctgag cgccgcagca gtggtgacac
      841 cagcagcaga cgctcacgga gaaacctcta acgccacaac agccggagcc gatcacggtt
      901 gcttccccca catcaaccac ggaaccgagc tgcagcacaa gatcgcagtg ggactccagt
      961 ggttcaccgt gatcgtggct atcgtgcagc tcatcttcta cggttggcac agcttcaagg
     1021 ccacaaccgg ctgggaggag gtctacgtct gcgtgatcga gctcgtcaag tgcttcatcg
     1081 agctgttcca cgaggtcgac agcccagcca cagtgtacca gaccaacgga ggagccgtga
     1141 tttggctgcg gtacagcatg tggctcctga cttgccccgt gatcctgatc cacctgagca
     1201 acctgaccgg actgcacgaa gagtacagca agcggaccat gaccatcctg gtgaccgaca
     1261 tcggcaacat cgtgtggggg atcacagccg cctttacaaa gggccccctg aagatcctgt
     1321 tcttcatgat cggcctgttc tacggcgtga cttgcttctt ccagatcgcc aaggtgtata
     1381 tcgagagcta ccacaccctg cccaaaggcg tctgccggaa gatttgcaag atcatggcct
     1441 acgtcttctt ctgctcttgg ctgatgttcc ccgtgatgtt catcgccgga cacgagggac
     1501 tgggcctgat cacaccttac accagcggaa tcggccacct gatcctggat ctgatcagca
     1561 agaacacttg gggcttcctg ggccaccacc tgagagtgaa gatccacgag cacatcctga
     1621 tccacggcga catccggaag acaaccacca tcaacgtggc cggcgagaac atggagatcg
     1681 agaccttcgt cgacgaggag gaggagggag gagtggcggc accggtagta gcagtgagta
     1741 agggcgagga agtgatcaaa gagttcatgc ggtttaaggt gagaatggaa ggaagcatga
     1801 acggccacga gttcgaaatt gagggagaag gagagggacg gccctacgag ggcacccaga
     1861 cagccaagct gaaagtgaca aagggcgggc ctctgccatt cgcttgggac atcctgagcc
     1921 cacagtttat gtacggctcc aaggcctatg tgaaacatcc agctgacatt cccgattata
     1981 agaaactgag cttccccgag gggtttaagt gggaaagagt gatgaacttc gaggacggag
     2041 gcctggtgac tgtgacccag gacagctccc tgcaggatgg gaccctgatc tacaaggtga
     2101 aaatgagagg gacaaatttt ccccctgatg gacctgtgat gcagaagaaa actatgggat
     2161 gggaggcctc caccgaaagg ctgtatccac gcgacggggt gctgaaagga gaaatccacc
     2221 aggctctgaa gctgaaagat gggggacatt acctggtgga gttcaagaca atctacatgg
     2281 ccaagaaacc tgtgcagctg ccaggctact attacgtgga cacaaaactg gatatcactt
     2341 cacacaacga ggactacact attgtggagc agtatgaacg gagcgagggg agacaccatc
     2401 tgttcctggg ccatgggact ggaagtaccg gctcagggtc tagtggaacc gcctcaagcg
     2461 aggataacaa tatggctgtg atcaaagagt tcatgaggtt taaggtgcgc atggagggca
     2521 gcatgaatgg gcacgaattt gagattgaag gagagggcga agggaggcct tacgagggca
     2581 cacagactgc caagctgaaa gtgaccaagg gaggaccact gcctttcgct tgggatatcc
     2641 tgtctcctca gtttatgtac ggaagtaagg cctatgtcaa gcatcccgct gacattcctg
     2701 attacaagaa actgtctttc ccagagggct ttaagtggga gagagtgatg aattttgaag
     2761 atggaggcct ggtgaccgtg acacaggact cctctctgca ggatggcact ctgatctaca
     2821 aagtcaaaat gcgcggcacc aattttccac ccgatgggcc cgtgatgcag aagaaaacaa
     2881 tggggtggga ggccagcact gaacggctgt atcctagaga cggagtgctg aagggcgaaa
     2941 tccaccaggc cctgaagctg aaagacggcg gccactacct ggtggagttc aaaaccatct
     3001 acatggccaa gaaaccagtg cagctgcccg gctattacta tgtggacacc aagctggata
     3061 tcacatccca caatgaagac tacaccattg tggaacagta tgagaggtct gaaggacgcc
     3121 accatctgtt tctgtacggc atggatgagc tgtataagta agaattcgat atcaagctta
     3181 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     3241 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     3301 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     3361 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     3421 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     3481 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3541 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     3601 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3661 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3721 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     3781 accgtcgacc cgggcggccg ctcataaagc tcgactgtgc cttctagttg ccagccatct
     3841 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3901 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3961 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     4021 gatgcggtcc ggactgtaca attgcattca ttttatgttt caggttcagg gggagatgtg
     4081 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatcttcc
     4141 tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac aaggaacccc
     4201 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     4261 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     4321 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4381 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4441 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4501 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4561 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4621 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4681 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4741 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4801 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4861 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4921 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4981 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5041 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5101 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5161 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5221 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5281 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5341 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5401 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5461 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5521 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5581 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5641 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5701 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5761 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5821 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5881 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5941 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6001 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6061 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6121 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6181 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6241 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6301 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     6361 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     6421 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     6481 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     6541 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     6601 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     6661 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     6721 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg
     6781 ctgcgcaac
//
