LOCUS       YY225:_pAAV_TRE::GFP-_G 5222 bp ds-DNA     circular     23-OCT-2024
DEFINITION  AAV expression of jRGECO1a, a red fluorescent calcium sensor
            protein, from the Synapsin promoter.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
COMMENT     Imported using the Genbank importer. File name:
            addgene-plasmid-100854-sequence-198193.gbk
FEATURES             Location/Qualifiers
     misc_feature    1..8
                     /label="NotI"
     polyA_signal    24..145
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(35..54)
                     /label="EBV-rev"
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     89..108
                     /label="SV40pA-R"
                     /note="SV40 polyA, reverse primer"
     repeat_region   307..436
                     /label="AAV2 ITR"
     primer_bind     complement(502..519)
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     rep_origin      complement(673..1261)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
                     /direction="LEFT"
     primer_bind     complement(753..772)
                     /label="pBR322ori-F"
                     /note="pBR322 origin, forward primer"
     CDS             complement(1432..2292)
                     /label="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, andrelated antibiotics"
                     /gene="bla"
                     /product="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"
     primer_bind     2055..2074
                     /label="Amp-R"
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2293..2397)
                     /label="AmpR promoter"
                     /gene="bla"
     rep_origin      complement(2423..2878)
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
                     /direction="LEFT"
     primer_bind     complement(2560..2581)
                     /label="F1ori-F"
                     /note="F1 origin, forward primer"
     primer_bind     2772..2791
                     /label="F1ori-R"
                     /note="F1 origin, reverse primer"
     repeat_region   2950..3079
                     /label="AAV2 ITR"
     misc_feature    3147..3152
                     /label="ApaI"
     protein_bind    3187..3205
                     /label="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
     protein_bind    3223..3241
                     /label="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
     protein_bind    3259..3277
                     /label="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
     protein_bind    3295..3313
                     /label="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
     protein_bind    3331..3349
                     /label="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
     protein_bind    3367..3385
                     /label="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
     protein_bind    3403..3421
                     /label="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
     primer_bind     3466..3490
                     /label="LNCX"
                     /note="Human CMV promoter, forward primer"
     misc_feature    3560..3565
                     /label="XbaI"
     CDS             4862..4867
                     /label="HindIII"
                     /note="codon_start: 1 note: mammalian codon-optimized product: EGFP destabilized by a mutated version of residues 422-461 of mouse ornithine decarboxylase transl_table: 1 translation: MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKKLSHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINV* modified by: DCLin-workstation"
     motif           4997..5005
                     /label="N1"
                     /note="created by: DCLin-workstation"
     motif           4997..5047
                     /label="N4"
                     /note="created by: DCLin-workstation"
     motif           5011..5019
                     /label="N1"
                     /note="created by: DCLin-workstation"
     extracted re... 5011..5024
                     /label="Extracted region from DL146_lenti_TRE_GFP-PAK4cat_P1N4"
                     /note="Extracted interval: 5079 -> 5092 Original Circular Sequence Length: 9952"
     motif           5025..5033
                     /label="N1"
                     /note="created by: DCLin-workstation"
     motif           5039..5047
                     /label="N1"
                     /note="created by: DCLin-workstation"
ORIGIN
        1 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
       61 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
      121 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
      181 tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat
      241 cgataaggat cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta
      301 actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
      361 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
      421 gcgagcgagc gcgcagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
      481 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
      541 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
      601 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
      661 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
      721 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
      781 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
      841 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
      901 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
      961 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1021 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1081 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     1141 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1201 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     1261 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     1321 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     1381 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     1441 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     1501 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     1561 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     1621 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     1681 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     1741 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     1801 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     1861 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     1921 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     1981 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     2041 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     2101 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     2161 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     2221 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     2281 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     2341 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     2401 atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta
     2461 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     2521 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     2581 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     2641 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     2701 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     2761 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     2821 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     2881 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     2941 ttacgccagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg
     3001 gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac
     3061 tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt atctacgtag
     3121 ccatgctcta ggaagatctc tgcagagggc ccccactagt ctttcgtctt caagaattcc
     3181 gtttactccc tatcagtgat agagaacgta tgaagagttt actccctatc agtgatagag
     3241 aacgtatgca gactttactc cctatcagtg atagagaacg tataaggagt ttactcccta
     3301 tcagtgatag agaacgtatg accagtttac tccctatcag tgatagagaa cgtatctaca
     3361 gtttactccc tatcagtgat agagaacgta tatccagttt actccctatc agtgatagag
     3421 aacgtataag ctttaggcgt gtacggtggg cgcctataaa agcagagctc gtttagtgaa
     3481 ccgtcagatc gcctggagca attccacaac acttttgtct tataccaact ttccgtacca
     3541 cttcctaccc tcgtaaaggt ctagaATGAG TAAAGGAGAA GAACTTTTCA CTGGAGTTGT
     3601 CCCAATTCTT GTTGAATTAG ATGGTGATGT TAATGGGTAC AAATTTTCTG TCAGTGGAGA
     3661 GGGTGAAGGT GATGCAACAT ACGGAAAACT TACCCTTAAA TTTATTTGCA CTACTGGAAA
     3721 ACTACCTGTT CCATGGCCAA CACTTGTCAC TACTCTCACT TATGGTGTTC AATGCTTTTC
     3781 AAGATATCCA GATCATATGA AGCGGCACGA CTTCTTCAAG AGCGCCATGC CTGAGGGATA
     3841 CGTGCAGGAG AGGACCATCT TCTTCAAGGA CGACGGGAAC TACAAGACAC GTGCTGAAGT
     3901 CAAGTTTGAG GGAGACACCC TCGTCAACAG GATCGAGCTT AAGGGAATCG ATTTCAAGGA
     3961 GGACGGAAAC ATCCTCGGCC ACAAGTTGGA ATACAACTAC AACTCCCACA ACGTATACAT
     4021 CATGGCCGAC AAGCAAAAGA ACGGCATCAA AGCCAACTTC AAGACCCGCC ACAACATCGA
     4081 AGACGGCGGC GTGCAACTCG CTGATCATTA TCAACAAAAT ACTCCAATTG GCGATGACCC
     4141 TGTCCTTTTA CCAGACAACC ATTACCTGTC CACACAATCT GCCCTTTCGA AAGATCCCAA
     4201 CGAAAAGAGA GACCACATGG TCATTCTTGA GTTTGTAACG GCTGCTGGGA TTACACATGG
     4261 CATGGATGAA CTATACAAAG GTGGATCTGG GGGTAGCGGA GGATCAGGGG GTTCCGGAGG
     4321 TTCTGGCGGA AGTAGCAAAG CGAAAATTGG CATTGTGACC GTGAGCGATC GCGCGAGCGC
     4381 GGGCATTACC GCAGATATTA GCGGCAAAGC CATTATTCTG GCGCTGAATC TGTATCTGAC
     4441 CAGCGAATGG GAACCGATTT ATCAGGTGAT TCCGGATGAA CAGAAAGTTA TTGAACGCAC
     4501 CCTGATTAAA ATGGCGGATA TTCAGGATTG TTGCCTGATT GTTACCACCG GCGGCACCGG
     4561 TCCGGCGAAA CGTGATGTGA CTCCTGAAGC AACTGAAGCG GTGTGCGATC GTATGATGCC
     4621 GGGCTTTGGC GAACTGATGC GCGCGGAAAG CCTGAAAGAA GTGCCGACCG CGATTCTGTC
     4681 TCGCCAGACT GCGGGTCTGA GAGGTGATAG CCTGATCGTG AACCTGCCGG GTGATCCGGC
     4741 CAGCATTAGC GATTGCCTGC TGGCAGTGTT TCCGGCAATT CCGTATTGCA TTGATCTGAT
     4801 GGAAGGGCCG TATCTGGAAT GCAACGAAGC GATGATCAAA CCGTTTCGCC CGAAAGCCAA
     4861 AAAGCTTAGC CATGGCTTCC CGCCGGCGGT GGCGGCGCAG GATGATGGCA CGCTGCCCAT
     4921 GTCTTGTGCC CAGGAGAGCG GGATGGACCG TCACCCTGCA GCCTGTGCTT CTGCTAGGAT
     4981 CAATGTGTGA TCTAGATTAT TTATTGATCC TTATTTATTG ATCCTTATTT ATTGATCCTT
     5041 ATTTATTGTC GGGGCGGCCG GCCGCTTCGA GCAGACATGA TAAGATACAT TGATGAGTTT
     5101 GGACAAACCA CAACTAGAAT GCAGTGAAAA AAATGCTTTA TTTGTGAAAT TTGTGATGCT
     5161 ATTGCTTTAT TTGTAACCAT TATAAGCTGC AATAAACAAG TTgatatcct cgagagatct
     5221 ac
//