LOCUS       Exported                5298 bp DNA     circular SYN 14-OCT-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5298)
  AUTHORS   Fishell Lab
  TITLE     Direct Submission
  JOURNAL   Exported Nov 3, 2021 from SnapGene 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5298
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    1..130
                     /label=ITR
     misc_feature    150..608
                     /label=hSyn1 Promoter
     misc_feature    609..628
                     /label=P/E cut
     misc_feature    609..622
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_G_EGFP-N"
     misc_feature    609..622
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_G_EGFP-N"
     misc_feature    623..628
                     /label=Gibs_Flex_Fb
     misc_feature    623..628
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     misc_feature    629..662
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     protein_bind    629..662
                     /label=LoxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    651..662
                     /label=Gibson_Flex_R1
     protein_bind    663..696
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    747..780
                     /label=F5
     misc_feature    781..782
                     /label=Changed gg to aa to remove AscI site
     misc_feature    792..825
                     /label=MCS
                     /note=" multiple cloning site
                     "
     primer_bind     810..854
                     /label=GA-dTom.Rev
     misc_feature    831..1544
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     misc_feature    831..1544
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     CDS             complement(831..1538)
                     /codon_start=1
                     /label=dTomato
                     /translation="MAVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQT
                     AKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDG
                     GLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEI
                     HQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGR
                     HHLFLYGMDELYK"
     misc_feature    831..1532
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     misc_feature    831..852
                     /label=Gibs_Fb
     primer_bind     831..852
                     /label=Gibs_JM1.1_F
     misc_feature    831..838
                     /label=Gibson_NLS_R2
     primer_bind     complement(837..855)
                     /label=Seq-YFP3.F
     primer_bind     complement(1513..1546)
                     /label=Gibs_JM1.1_R
     misc_feature    1513..1532
                     /label=Gibs_Ra
     primer_bind     complement(1528..1564)
                     /label=GA-dTom.For
     misc_feature    1533..1535
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     misc_feature    1536..1544
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     misc_feature    1539..1544
                     /label=Kozak
     misc_feature    1550..1568
                     /label=MCS
                     /note=" multiple cloning site
                     "
     protein_bind    complement(1579..1612)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    complement(1663..1696)
                     /label=F5
     misc_feature    1697..1730
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     protein_bind    1697..1730
                     /label=LoxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    1719..1730
                     /label=Gibson_Flex_R1
     misc_feature    1731..1738
                     /label=AscI
     misc_feature    1739..2327
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     polyA_signal    2335..2559
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2561..2701
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      2776..3231
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3513..3617
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3618..4478
                     /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      4649..5237
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gctagactgc agagggccct gcgtatgagt gcaagtgggt tttaggacca
      181 ggatgaggcg gggtgggggt gcctacctga cgaccgaccc cgacccactg gacaagcacc
      241 caacccccat tccccaaatt gcgcatcccc tatcagagag ggggagggga aacaggatgc
      301 ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac agtgccttcg cccccgcctg
      361 gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc gccggtcccc
      421 cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc cgccgccggc ccagccggac
      481 cgcaccacgc gaggcgcgag ataggggggc acgggcgcga ccatctgcgc tgcggcgccg
      541 gcgactcagc gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag
      601 cgcagtcaga tatcgtttaa actccggaat aacttcgtat agcatacatt atacgaagtt
      661 atgaagttcc tattctctag aaagtatagg aacttcgcag aatggtagct ggattgtagc
      721 tgctattagc aatatgaaac ctcttagaag ttcctattct tcaaaaggta taggaacttc
      781 aacgcgccct agaattcgag ctcggtaccc ggggatcctc tagagtcgac ttacttgtac
      841 agctcgtcca tgccgtacag gaacaggtgg tggcggccct cggagcgctc gtactgttcc
      901 acgatggtgt agtcctcgtt gtgggaggtg atgtccagct tggtgtccac gtagtagtag
      961 ccgggcagtt gcacgggctt cttggccatg tagatggtct tgaactccac caggtagtgg
     1021 ccgccgtcct tcagcttcag ggcctggtgg atctcgccct tcagcacgcc gtcgcggggg
     1081 tacaggcgct cggtggaggc ctcccagccc atggtcttct tctgcattac ggggccgtcg
     1141 ggggggaagt tggtgccgcg catcttcacc ttgtagatca gcgtgccgtc ctgcagggag
     1201 gagtcctggg tcacggtcac cagaccgccg tcctcgaagt tcatcacgcg ctcccacttg
     1261 aagccctcgg ggaaggacag cttcttgtaa tcggggatgt cggcggggtg cttcacgtac
     1321 gccttggagc cgtacatgaa ctggggggac aggatgtccc aggcgaaggg cagggggccg
     1381 cccttggtca ccttcagctt ggcggtctgg gtgccctcgt aggggcggcc ctcgccctcg
     1441 ccctcgatct cgaactcgtg gccgttcatg gagccctcca tgcgcacctt gaagcgcatg
     1501 aactctttga tgacctcctc gcccttgctc acggccatgg tggcgtcgac ctgcaggcat
     1561 gcaagcttgg tggctagcga agttcctata ctttctagag aataggaact tcttgcctta
     1621 acccagaaat tatcactgtt attctttaga atggtgcaaa gagaagttcc tatacctttt
     1681 gaagaatagg aacttcataa cttcgtatag catacattat acgaagttat ggcgcgccaa
     1741 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     1801 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     1861 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     1921 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     1981 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     2041 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2101 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     2161 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     2221 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     2281 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gatcctgtgc
     2341 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     2401 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     2461 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     2521 acaatagcag gcatgctggg gatgcggtgg gctctatggc aggaacccct agtgatggag
     2581 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     2641 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     2701 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     2761 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2821 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2881 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2941 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     3001 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3061 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct
     3121 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3181 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     3241 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     3301 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     3361 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     3421 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     3481 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3541 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3601 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3661 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3721 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3781 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3841 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3901 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3961 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4021 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4081 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4141 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4201 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     4261 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4321 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4381 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4441 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4501 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4561 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4621 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4681 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4741 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     4801 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4861 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4921 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4981 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5041 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5101 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5161 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5221 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5281 ggccttttgc tcacatgt
//
