LOCUS       Exported                7455 bp DNA     circular SYN 12-JUL-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7455)
  AUTHORS   Fishell Lab
  TITLE     Direct Submission
  JOURNAL   Exported Aug 19, 2021 from SnapGene Viewer 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7455
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    1..130
                     /label=ITR
     misc_feature    131..608
                     /label=hSyn1 Promoter
     primer_bind     156..179
                     /label=N2c_Frecombineddel
     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=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     misc_feature    623..628
                     /label=Gibs_Flex_Fb
     protein_bind    629..662
                     /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    664..801
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     misc_feature    664..676
                     /label=Gibs_Flex_Fb
     protein_bind    669..702
                     /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."
     protein_bind    753..786
                     /label=LoxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    775..794
                     /label=Gibson_Flex_R1
     primer_bind     786..828
                     /label=GP9-Gib-F
     misc_feature    795..800
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     misc_feature    795..800
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_G_EGFP-N"
     misc_feature    795..800
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     misc_feature    795..800
                     /label=SpeI
     CDS             complement(807..2381)
                     /codon_start=1
                     /label=N2cG
                     /translation="MVPQVLLFVLLLGFSLCFGKFPIYTIPDELGPWSPIDIHHLSCPN
                     NLVVEDEGCTNLSEFSYMELKVGYISAIKVNGFTCTGVVTEAETYTNFVGYVTTTFKRK
                     HFRPTPDACRAAYNWKMAGDPRYEESLHNPYPDYHWLRTVRTTKESLIIISPSVTDLDP
                     YDKSLHSRVFPGGKCSGITVSSTYCSTNHDYTIWMPENPRPRTPCDIFTNSRGKRASNG
                     NKTCGFVDERGLYKSLKGACRLKLCGVLGLRLMDGTWVAMQTSDETKWCPPDQLVNLHD
                     FRSDEIEHLVVEELVKKREECLDALESIMTTKSVSFRRLSHLRKLVPGFGKAYTIFNKT
                     LMEADAHYKSVRTWNEIIPSKGCLKVGGRCHPHVNGVFFNGIILGPDDHVLIPEMQSSL
                     LQQHMELLESSVIPLMHPLADPSTVFKEGDEAEDFVEVHLPDVYKQISGVDLGLPNWGK
                     YVLMTAGAMIGLVLIFSLMTWCRRANRPESKQRSFGGTGGNVSVTSQSGKVIPSWESYK
                     SGGEIRL"
     CDS             complement(2382..2438)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(2455..3164)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDE"
     CDS             complement(3165..3221)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(3230..3699)
                     /codon_start=2
                     /label=TVA
                     /translation="ARLLPALLLLLLPGNVTGNGSGNGSLSRCPPGQFRCSEPPGAHGE
                     CYPQDWLCDGHPDCDDGRDEWGCGTSAIPAVPTDNGTEAPTAPAPGRALPARNHGRMWM
                     LITAVLLCCLVAVGGIAAWGKSKAKSRSDIFSLESASKELLVPDKSQADLFS"
     primer_bind     3372..3395
                     /label=N2c_Fcorrect
     primer_bind     complement(3682..3710)
                     /label=GP9-Gib-R
     misc_feature    3712..3853
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     misc_feature    3717..3723
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_G_EGFP-N"
     misc_feature    3717..3723
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     misc_feature    3718..3723
                     /label=SpeI
     misc_feature    3736..3769
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     protein_bind    complement(3736..3769)
                     /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."
     misc_feature    3762..3769
                     /label=Gibs_Flex_Fb
     misc_feature    3820..3853
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     protein_bind    complement(3820..3853)
                     /label=LoxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3820..3831
                     /label=Gibson_Flex_R1
     protein_bind    3854..3887
                     /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    3896..4484
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3963..3983)
                     /label=Primer Rcommon
     polyA_signal    4492..4716
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4718..4858
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4933..5388
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5670..5774
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5775..6635
                     /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      6806..7394
                     /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 actccggaga agttcctatt ctctagaaag tataggaact
      661 tcaccggaat aacttcgtat aggatacttt atacgaagtt atgcagaatg gtagctggat
      721 tgtagctgct attagcaata tgaaacctct taataacttc gtatagcata cattatacga
      781 agttataacg cgccactagt gaattcttac agtctgatct cacctccact cttatatgat
      841 tcccatgaag gtatgacttt tccgctttgg gaagtgactg acacattccc ccctgtccct
      901 ccaaaactgc gttgtttcga ttctggtcga ttggctcttc tgcaccatgt cattagggaa
      961 aatatcaaca ccaggccaat catggcccct gcagtcatca atacatactt tccccagttc
     1021 gggagaccca ggtcaacccc tgagatctgt ttgtacacat cggggaggtg aacttcaaca
     1081 aaatcctcag cctcatcacc ttctttgaaa actgtagaag ggtctgccag ggggtgcatc
     1141 agggggataa ctgaagattc caacaactcc atatgttgct ggaggaggga tgattgcatc
     1201 tctgggatta ggacatggtc gtcaggccct aatattatac cattgaaaaa caccccgttc
     1261 acatgaggat ggcaccttcc tccaactttc aaacaccctt ttgaggggat gatctcattc
     1321 caggtccgga ctgacttgta gtgagcatca gcctccatca aggttttgtt gaatatggta
     1381 tatgcttttc caaaccctgg gacaagtttt ctcaggtgac tgagacgtct gaaacttact
     1441 gacttggtgg tcatgatgga ctctaatgca tccagacatt cctctctttt cttgactaac
     1501 tcctccacaa cgagatgctc aatctcgtct gagcgaaagt cgtgcaaatt caccaactga
     1561 tctggagggc accatttggt ctcatctgat gtttgcatcg cgacccatgt tccatccata
     1621 agtctaagtc caagaactcc acataacttg agcctgcatg ctccttttag agacttatac
     1681 aggcctcttt catccacaaa gccgcaagtc ttgttcccgt tggatgctct cttccctctg
     1741 ctattggtaa aaatgtcaca aggtgtcctt ggtctcggat tctcgggcat ccaaatggtg
     1801 taatcatggt tagttgagca gtaggtagag gacaccgtta ttcctgagca ctttccgcca
     1861 gggaagaccc ttgagtgaag ggatttgtca tatgggtcca aatctgtcac acttggggat
     1921 atgataatga gggactcttt ggtggttctt acagttcgaa gccagtggta gtcggggtat
     1981 ggattgtgta gggactcttc atatctgggg tcaccggcca tcttccagtt atacgcggct
     2041 ctacatgcgt ctggggtggg gcggaaatgc tttctcttga atgtggttgt gacataacca
     2101 acaaagttgg tgtaggtctc tgcctctgtc acaacacctg tgcaagtgaa cccgttcact
     2161 ttgatggctg agatgtatcc cactttgagt tccatgtagg agaactcgga caggttggta
     2221 catccttcat cctccacaac caggttattt ggacagctga gatggtgtat gtcaataggg
     2281 ctccagggac caagttcgtc tggtatcgtg taaatgggga acttcccgaa acacaacgaa
     2341 aaacccagaa ggagtacaaa caaaagaacc tgaggaacca taggtccagg gttctcctcc
     2401 acgtctccag cctgcttcag caggctgaag ttagtagctc cgcttccctt gtacagctcg
     2461 tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc
     2521 ttctcgttgg ggtctttgct cagggcggac tgggtgctca ggtagtggtt gtcgggcagc
     2581 agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg
     2641 ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg
     2701 gccatgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg
     2761 ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg
     2821 aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc
     2881 tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg
     2941 tagcggctga agcactgcac gccgtaggtc agggtggtca cgagggtggg ccagggcacg
     3001 ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccgtaggt ggcatcgccc
     3061 tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg
     3121 atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatgggtcc agggttctcc
     3181 tccacgtctc cagcctgctt cagcaggctg aagttagtag ctccggatcc ggagaacaag
     3241 tctgcctggc tcttgtcagg caccagcagc tccttggatg cgctttcaag actgaagatg
     3301 tcagacctgc tttttgcttt ggacttcccc catgcagcga taccacccac agctaccagg
     3361 cagcacagga gcactgcagt gatcagcatc cacatgcggc cgtgattcct ggctggcaga
     3421 gcacgtccag gagcaggggc agtgggagcc tctgtgccgt tgtccgtggg caccgcgggg
     3481 atcgcgctgg tcccgcagcc ccactcgtcc cgcccgtcgt cgcagtcggg gtgtccgtcg
     3541 cacagccagt cctgcgggta acactccccg tgggcaccgg gcggctccga gcagcggaac
     3601 tgaccggggg ggcaacggga caaagaaccg ttaccggacc cgttaccggt cacgttaccg
     3661 ggcagcagca gcagcagcag cgcgggcagc agccgcgcca tggtggcggc aagctttact
     3721 agtggtggcg ctagcataac ttcgtataaa gtatcctata cgaagttatt tgccttaacc
     3781 cagaaattat cactgttatt ctttagaatg gtgcaaagaa taacttcgta taatgtatgc
     3841 tatacgaagt tatgaagttc ctattctcta gaaagtatag gaacttcggc gcgccaatca
     3901 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     3961 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     4021 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     4081 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     4141 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     4201 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     4261 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     4321 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     4381 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     4441 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggat cctgtgcctt
     4501 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     4561 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     4621 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     4681 atagcaggca tgctggggat gcggtgggct ctatggcagg aacccctagt gatggagttg
     4741 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     4801 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     4861 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     4921 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4981 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     5041 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5101 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     5161 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5221 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt
     5281 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     5341 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     5401 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     5461 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     5521 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     5581 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     5641 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5701 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5761 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5821 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5881 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5941 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     6001 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     6061 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     6121 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     6181 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     6241 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6301 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     6361 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     6421 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6481 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6541 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6601 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     6661 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6721 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6781 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6841 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6901 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     6961 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7021 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7081 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7141 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7201 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7261 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7321 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7381 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7441 cttttgctca catgt
//
