LOCUS       Exported                7197 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Can be used to generate AAV virus that will express tdTomato in the 
            presence of Cre.
ACCESSION   .
VERSION     .
KEYWORDS    AAV pCAG-FLEX-tdTomato-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7197)
  AUTHORS   Oh SW, Harris JA, Ng L, Winslow B, Cain N, Mihalas S, Wang Q, Lau C,
            Kuan L, Henry AM, Mortrud MT, Ouellette B, Nguyen TN, Sorensen SA, 
            Slaughterbeck CR, Wakeman W, Li Y, Feng D, Ho A, Nicholas E, 
            Hirokawa KE, Bohn P, Joines KM, Peng H, Hawrylycz MJ, Phillips JW, 
            Hohmann JG, Wohnoutka P, Gerfen CR, Koch C, Bernard A, Dang C, Jones
            AR, Zeng H
  TITLE     A mesoscale connectome of the mouse brain.
  JOURNAL   Nature. 2014 Apr 10;508(7495):207-14. doi: 10.1038/nature13186. Epub
            2014 Apr 2.
  PUBMED    24695228
REFERENCE   2  (bases 1 to 7197)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7197
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     enhancer        234..537
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        539..816
                     /label=chicken beta-actin promoter
     intron          818..1835
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1759..1781
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1843..1862
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1939..1972
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    1983..2016
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /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 (Lee and 
                     Saito, 1988)."
     primer_bind     complement(2845..2868)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(3043..3062)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3312..3330
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3460..3469
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    complement(3500..3533)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    complement(3544..3577)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /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 (Lee and 
                     Saito, 1988)."
     misc_feature    3601..4189
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3654..3674)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4072..4083)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4224..4241)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4230..4437
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4460..4600
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4460..4589
                     /label=AAV2 ITR
     rep_origin      4675..5130
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4762..4781)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4972..4993
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5147..5166)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5266..5288
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5326..5344)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5412..5516
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5517..6377
                     /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     complement(5735..5754)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6548..7136
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7037..7056
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgaaacaa ttctgcagga atctagttat
      181 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      241 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      301 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      361 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      421 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      481 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
      541 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
      601 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg gggggggggg
      661 cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg
      721 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
      781 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggga gtcgctgcga cgctgccttc
      841 gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt
      901 tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg
      961 tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg
     1021 gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg
     1081 ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt
     1141 gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg
     1201 ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg
     1261 gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc
     1321 acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg
     1381 gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg
     1441 gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc
     1501 gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa
     1561 tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga
     1621 agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc
     1681 gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg
     1741 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg
     1801 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg
     1861 tgctgtctca tcattttggc aaagaattga tttgataccg cgggccccgc gtggacggta
     1921 cccttcaggt cgacgtcgat aacttcgtat agcatacatt atacgaagtt atccgaatcg
     1981 caataacttc gtataggata ctttatacga agttatttgg cgcgccgcta gcttacttgt
     2041 acagctcgtc catgccgtac agaaacagat ggtgcctccc ctcgctccgc tcatactgct
     2101 ccacaatggt gtagtcttcg ttatggctag taatatccag ctttgtgtcg acgtaatagt
     2161 agccaggcag ctgcacgggt ttcttggcca tgtagatggt tttgaattcg accagataat
     2221 gccctccatc tttcagcttc agtgcctggt ggatctctcc cttcagcacg ccgtctctgg
     2281 ggtacagcct ttctgttgaa gcctcccagc ccattgtctt cttctgcatg actgggccat
     2341 cagggggaaa attggtgccg cgcattttca ccttatagat cagagtgccg tcctgcagag
     2401 agctgtcctg ggtgacagtc accaggcccc cgtcttcgaa gttcatcact ctttcccact
     2461 taaacccctc tgggaagctc agtttcttgt aatctggaat gtcagcaggg tgtttcacgt
     2521 aggctttact tccatacata aactgaggag acaggatgtc ccatgcgaat ggcagagggc
     2581 cgccctttgt cactttcagc tttgctgtct gggtgccctc atagggcctc ccttccccct
     2641 ctccttcaat ctcaaactcg tgcccgttca tgctgccctc catcctgact ttaaacctca
     2701 taaactcctt aatcacggcc atgttgttgt cctcggagga ggcggtgccg gagctgccgc
     2761 tgccggtgct gccggtacca tgccccagga acaggtggtg gcggccctcg gagcgctcgt
     2821 actgttccac gatggtgtag tcctcgttgt gggaggtgat gtccagcttg gtgtccacgt
     2881 agtagtagcc gggcagttgc acgggcttct tggccatgta gatggtcttg aactccacca
     2941 ggtagtggcc gccgtccttc agcttcaggg cctggtggat ctcgcccttc agcacgccgt
     3001 cgcgggggta caggcgctcg gtggaggcct cccagcccat ggtcttcttc tgcattacgg
     3061 ggccgtcggg ggggaagttg gtgccgcgca tcttcacctt gtagatcagc gtgccgtcct
     3121 gcagggagga gtcctgggtc acggtcacca gaccgccgtc ctcgaagttc atcacgcgct
     3181 cccacttgaa gccctcgggg aaggacagct tcttgtaatc ggggatgtcg gcggggtgct
     3241 tcacgtacgc cttggagccg tacatgaact ggggggacag gatgtcccag gcgaagggca
     3301 gggggccgcc cttggtcacc ttcagcttgg cggtctgggt gccctcgtag gggcggccct
     3361 cgccctcgcc ctcgatctcg aactcgtggc cgttcatgga gccctccatg cgcaccttga
     3421 agcgcatgaa ctctttgatg acctcctcgc ccttgctcac catggtggcg ctagcggcgc
     3481 gccaacctgg ccggccttga taacttcgta taatgtatgc tatacgaagt tatccgaatc
     3541 gcaataactt cgtataaagt atcctatacg aagttatcga agatcatcaa gcttatcgat
     3601 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     3661 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3721 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     3781 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     3841 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     3901 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3961 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     4021 gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     4081 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4141 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga
     4201 gcgctgctcg agagatcgat ctgcctcgac tgtgccttct agttgccagc catctgttgt
     4261 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     4321 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     4381 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggacac
     4441 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     4501 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     4561 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     4621 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     4681 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4741 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4801 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4861 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     4921 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4981 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga
     5041 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5101 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     5161 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     5221 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     5281 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     5341 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     5401 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5461 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     5521 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5581 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5641 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5701 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     5761 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5821 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     5881 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     5941 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6001 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6061 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6121 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6181 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6241 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6301 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6361 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     6421 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6481 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6541 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6601 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6661 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     6721 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6781 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6841 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6901 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6961 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7021 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7081 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7141 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
