LOCUS       Exported                5717 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 5717)
  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..5717
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    1..130
                     /label=ITR
     misc_feature    150..1041
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_G_EGFP-N"
     primer_bind     151..188
                     /label=Human_Dlx_F_Gibson
     misc_feature    151..166
                     /label=Gibson_hDlx_P_F
     misc_feature    166
                     /label=""A"" after PacI digest
     misc_feature    167..1005
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_G_EGFP-N"
     misc_feature    167..707
                     /label=hDlx
     misc_feature    167..370
                     /label=h56ii_enhancer
     misc_feature    167..188
                     /label=Gibson_hDlx_I_F
     misc_feature    complement(336..621)
                     /label=hI56i from Zerucha et al., 2000 in reverse
                     /note="100% identical to mouse homologous seq85%identical 
                     to zebrafish homologous seq
                     "
     primer_bind     complement(346..386)
                     /label=Human_Dlx_R_Gibson
     misc_feature    346..370
                     /label=Gibson_hDlx_I_R
     misc_feature    370
                     /label=h cloning
     misc_feature    371..704
                     /label=m56ii_enhancer
     misc_feature    371..386
                     /label=Gibson_hDlx_P_R
     misc_feature    371
                     /label=""C"" after PpumI digest
     primer_bind     514..533
                     /label=Seq_AAV_hDlx_F2
     primer_bind     complement(514..533)
                     /label=AAV_hDlx_R2
     promoter        740..788
                     /label=pB-Globin
     primer_bind     complement(743..762)
                     /label=AAV_hDlx_R1
     misc_feature    835..1005
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     intron          873..1005
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=Seq_AAV_hDlx_F3
     misc_feature    1006..1047
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     primer_bind     1012..1031
                     /label=Seq_AAV_Dlx_F
     misc_feature    1023..1042
                     /label=Gibson_NLS_F1a
     misc_feature    1026..1041
                     /label=Gibson_F_16bp
     misc_feature    1048..1081
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     protein_bind    1048..1081
                     /label=LoxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    1070..1081
                     /label=Gibson_Flex_R1
     protein_bind    1082..1115
                     /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    1166..1199
                     /label=F5
     misc_feature    1200..1201
                     /label=Changed gg to aa to remove AscI site
     misc_feature    1211..1244
                     /label=MCS
                     /note=" multiple cloning site
                     "
     primer_bind     1229..1273
                     /label=GA-dTom.Rev
     misc_feature    1250..1963
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     misc_feature    1250..1963
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     CDS             complement(1250..1957)
                     /codon_start=1
                     /label=dTomato
                     /translation="MAVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQT
                     AKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDG
                     GLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEI
                     HQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGR
                     HHLFLYGMDELYK"
     misc_feature    1250..1951
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     primer_bind     1250..1271
                     /label=Gibs_JM1.1_F
     misc_feature    1250..1257
                     /label=Gibson_NLS_R2
     primer_bind     complement(1256..1274)
                     /label=Seq-YFP3.F
     primer_bind     complement(1932..1965)
                     /label=Gibs_JM1.1_R
     primer_bind     complement(1947..1983)
                     /label=GA-dTom.For
     misc_feature    1952..1954
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     misc_feature    1955..1963
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     misc_feature    1958..1963
                     /label=Kozak
     misc_feature    1969..1987
                     /label=MCS
                     /note=" multiple cloning site
                     "
     protein_bind    complement(1998..2031)
                     /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(2082..2115)
                     /label=F5
     misc_feature    2116..2149
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     protein_bind    2116..2149
                     /label=LoxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2138..2149
                     /label=Gibson_Flex_R1
     misc_feature    2150..2157
                     /label=AscI
     misc_feature    2158..2746
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     polyA_signal    2754..2978
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2980..3120
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      3195..3650
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3932..4036
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4037..4897
                     /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      5068..5656
                     /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 agagggcccg cggccgcacg cgtttattca gaatgttatg
      181 cactcacagt ggtttggcat gcatctggtg aatttttttt aacgaaaaat tagtgttggt
      241 ttcgatgtat ggtagcattc tccctaacgt aatttgaata attcagcaaa gccccactac
      301 cagctgtact tctgcagcct cttccattct tttcagcatt ataattttgg ttaattttca
      361 attttaggtc ctacgtctct gcaatttgtg tatgaataac agaataattt ccctcttttg
      421 tttcgccttt cctgttcctg aatctaaata aagatggctt tttagtatta aaagtggaag
      481 aaaattacag gtaattatct ttgacggtaa aaacgctgta atcagcgggc tacatgaaaa
      541 attactctaa ttatggctgc atttaagaga atggaaaaaa accttcttgt ggataaaaac
      601 cttaaattgt ccccaatgtc tgcttcaaat tggatggcac tgcagctgga ggctttgttc
      661 agaattgatc ctggggagct acgaacccaa agtttcacag taggaagcta gatatcgttt
      721 aaacttcctg cagcccgggc tgggcataaa agtcagggca gagccatcta ttgcttacat
      781 ttgcttctag cctgcaagtc gaggagcgca gccttccaga agcagagcgc ggcgccttaa
      841 gctgcagaag ttggtcgtga ggcactgggc acgtaagtat caaggttaca agacaggttt
      901 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
      961 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt
     1021 tcaattacag ctcttaagaa agctagcata acttcgtata gcatacatta tacgaagtta
     1081 tgaagttcct attctctaga aagtatagga acttcgcaga atggtagctg gattgtagct
     1141 gctattagca atatgaaacc tcttagaagt tcctattctt caaaaggtat aggaacttca
     1201 acgcgcccta gaattcgagc tcggtacccg gggatcctct agagtcgact tacttgtaca
     1261 gctcgtccat gccgtacagg aacaggtggt ggcggccctc ggagcgctcg tactgttcca
     1321 cgatggtgta gtcctcgttg tgggaggtga tgtccagctt ggtgtccacg tagtagtagc
     1381 cgggcagttg cacgggcttc ttggccatgt agatggtctt gaactccacc aggtagtggc
     1441 cgccgtcctt cagcttcagg gcctggtgga tctcgccctt cagcacgccg tcgcgggggt
     1501 acaggcgctc ggtggaggcc tcccagccca tggtcttctt ctgcattacg gggccgtcgg
     1561 gggggaagtt ggtgccgcgc atcttcacct tgtagatcag cgtgccgtcc tgcagggagg
     1621 agtcctgggt cacggtcacc agaccgccgt cctcgaagtt catcacgcgc tcccacttga
     1681 agccctcggg gaaggacagc ttcttgtaat cggggatgtc ggcggggtgc ttcacgtacg
     1741 ccttggagcc gtacatgaac tggggggaca ggatgtccca ggcgaagggc agggggccgc
     1801 ccttggtcac cttcagcttg gcggtctggg tgccctcgta ggggcggccc tcgccctcgc
     1861 cctcgatctc gaactcgtgg ccgttcatgg agccctccat gcgcaccttg aagcgcatga
     1921 actctttgat gacctcctcg cccttgctca cggccatggt ggcgtcgacc tgcaggcatg
     1981 caagcttggt ggctagcgaa gttcctatac tttctagaga ataggaactt cttgccttaa
     2041 cccagaaatt atcactgtta ttctttagaa tggtgcaaag agaagttcct ataccttttg
     2101 aagaatagga acttcataac ttcgtatagc atacattata cgaagttatg gcgcgccaat
     2161 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     2221 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     2281 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     2341 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     2401 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     2461 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     2521 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     2581 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     2641 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     2701 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg atcctgtgcc
     2761 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     2821 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     2881 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     2941 caatagcagg catgctgggg atgcggtggg ctctatggca ggaaccccta gtgatggagt
     3001 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     3061 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     3121 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     3181 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3241 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3301 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3361 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     3421 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3481 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta
     3541 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3601 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     3661 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     3721 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     3781 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     3841 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     3901 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3961 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4021 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4081 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4141 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4201 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4261 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4321 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4381 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4441 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4501 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4561 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     4621 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4681 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4741 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4801 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4861 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4921 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4981 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5041 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5101 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5161 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     5221 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5281 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5341 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5401 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5461 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5521 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5581 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5641 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5701 gccttttgct cacatgt
//
