LOCUS       Exported                6826 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 6826)
  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..6826
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    1..130
                     /label=ITR
     misc_feature    150..608
                     /label=hSyn1 Promoter
     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"
     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    663..1527
                     /label=Stop
     polyA_signal    708..756
                     /note="synthetic polyadenylation signal"
     polyA_signal    765..1241
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     polyA_signal    1320..1368
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    1528..1561
                     /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    1563..1733
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     misc_feature    1563..1575
                     /label=Gibs_Flex_Fb
     protein_bind    1568..1601
                     /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    1652..1685
                     /label=LoxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    1674..1693
                     /label=Gibson_Flex_R1
     misc_feature    1694..1699
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     misc_feature    1694..1699
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_G_EGFP-N"
     misc_feature    1694..1699
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     misc_feature    1694..1699
                     /label=SpeI
     misc_feature    1700..1733
                     /label=MCS
                     /note=" multiple cloning site
                     "
     primer_bind     1718..1760
                     /label=GA-GCaMP6f.Rev
     misc_feature    1738..3097
                     /label=Insert_2
     CDS             complement(1742..3091)
                     /codon_start=1
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     primer_bind     2701..2720
                     /label=EGFP.R
                     /note="20 mer	60"
     primer_bind     complement(2759..2776)
                     /label=EGFP.F
                     /note="18 mer	61"
     primer_bind     complement(3078..3117)
                     /label=GA-GCaMP6f.For
     misc_feature    3092..3097
                     /label=Kozak
     misc_feature    3103..3258
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     misc_feature    3103..3121
                     /label=MCS
                     /note=" multiple cloning site
                     "
     misc_feature    3122..3128
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_G_EGFP-N"
     misc_feature    3122..3128
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     misc_feature    3123..3128
                     /label=SpeI
     misc_feature    3141..3174
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     protein_bind    complement(3141..3174)
                     /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    3167..3174
                     /label=Gibs_Flex_Fb
     misc_feature    3225..3258
                     /label=140708_Sequencing_Clone1
                     /note="Check 140708_AAV_GFP_1_R_140619_Seq_AAV_Dlx_R"
     protein_bind    complement(3225..3258)
                     /label=LoxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3225..3236
                     /label=Gibson_Flex_R1
     misc_feature    3259..3266
                     /label=AscI
     misc_feature    3267..3855
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     polyA_signal    3863..4087
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4089..4229
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4304..4759
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5041..5145
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5146..6006
                     /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      6177..6765
                     /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 tccctcagca ccatggctag cggcagcctc ggagtttgaa tagatagaat aaaatatctt
      721 tattttcatt ccatctgtgt gttggttttt tgtgtgagat ctacgggtgg catccctgtg
      781 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg
      841 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg
      901 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg
      961 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg
     1021 cctcctgggt tcaagcgatt ctcctgcctc aggctcccga gttgttggga ttccaggcat
     1081 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg
     1141 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc
     1201 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgcctcag ctaattgagt
     1261 aggggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     1321 ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg
     1381 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     1441 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     1501 cagccaacgt cggggcggca ggccgtggaa gttcctattc tctagaaagt ataggaactt
     1561 caccggaata acttcgtata ggatacttta tacgaagtta tgcagaatgg tagctggatt
     1621 gtagctgcta ttagcaatat gaaacctctt aataacttcg tatagcatac attatacgaa
     1681 gttataacgc gccactagtg aattcgagct cggtacccgg ggatcctcta gagtcgactc
     1741 acttcgctgt catcatttgt acaaactctt cgtagtttac ctgaccatcc ccatcgatgt
     1801 ctgcttccct gatcatttca tcaacctctt catctgttaa cttctctcca aggtttgtca
     1861 tcacgtggcg aagctctgct gcactgatgt agccattgcc atccttatca aacacaccga
     1921 acgcttctct aatttcttct tccgtgtccc tgtatttcat ttttcttgcc atcattgtca
     1981 ggaactcagg gaagtcgatt gtgccgtcac cgtcggcatc tacttcattg atcatgtcct
     2041 gcagctctgc ttctgtgggg ttctgcccca gagaccgcat caccgtcccc agctccttgg
     2101 ttgttattgt cccatccccg tccttgtcaa atagggagaa ttcctcttta aattctgcga
     2161 tctgctcttc agtcagttgg tccggcaggt tgtactccag cttgtgcccc aggatgttgc
     2221 cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga
     2281 acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct
     2341 ggatgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt
     2401 agcggctgaa gcactgcacg ccgtaggtca gggtggtcac gagggtgggc cagggcacgg
     2461 gcagcttgcc ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct
     2521 caccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga
     2581 tgggcaccac cccggtgaac agctcctcgc ccttgctcac catgctccct ccggtaccgc
     2641 ccttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     2701 ccatgtgatc gcgcttctcg ttggggtctt tcgaaagttt ggactgcacg ctcaggtagt
     2761 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtagg
     2821 cgagctgcac gccgccgtcc tcgatgttgt ggcggatctt gaagttcgcc ttgatgccgt
     2881 tcttctgctt gtcggccttg atatagacgt tctcgagtga gctcagccga cctatagctc
     2941 tgactgcgtg acctgtctta ttccacttac gacgtgatga gtcgaccatg gtggcgagat
     3001 ccttatcgtc atcgtcgtac agatcccgac ccatttgctg tccaccagtc atgctagcca
     3061 taccatgatg atgatgatga tgagaaccca tggtggcgtc gacctgcagg catgcaagct
     3121 ttactagtgg tggcgctagc ataacttcgt ataaagtatc ctatacgaag ttatttgcct
     3181 taacccagaa attatcactg ttattcttta gaatggtgca aagaataact tcgtataatg
     3241 tatgctatac gaagttatgg cgcgccaatc aacctctgga ttacaaaatt tgtgaaagat
     3301 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3361 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     3421 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     3481 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     3541 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     3601 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     3661 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     3721 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     3781 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3841 gggccgcctc cccgcctgga tcctgtgcct tctagttgcc agccatctgt tgtttgcccc
     3901 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3961 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     4021 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     4081 tctatggcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     4141 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     4201 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     4261 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     4321 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4381 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4441 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4501 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4561 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4621 ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     4681 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     4741 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     4801 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4861 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4921 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     4981 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     5041 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5101 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5161 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5221 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5281 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5341 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5401 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5461 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5521 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5581 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5641 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5701 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5761 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5821 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5881 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5941 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6001 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6061 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6121 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6181 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6241 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6301 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     6361 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6421 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6481 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6541 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6601 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6661 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6721 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6781 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
