LOCUS       Exported                4629 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV vector to drive TET-ON activator expression from human synapsin 
            I promoter.
ACCESSION   .
VERSION     .
KEYWORDS    AAV-hSynI-rtTAV16
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4629)
  AUTHORS   Watakabe A, Takaji M, Kato S, Kobayashi K, Mizukami H, Ozawa K, 
            Ohsawa S, Matsui R, Watanabe D, Yamamori T
  TITLE     Simultaneous visualization of extrinsic and intrinsic axon 
            collaterals in Golgi-like detail for mouse corticothalamic and 
            corticocortical cells: a double viral infection method.
  JOURNAL   Front Neural Circuits. 2014 Sep 17;8:110. doi: 
            10.3389/fncir.2014.00110. eCollection 2014.
  PUBMED    25278843
REFERENCE   2  (bases 1 to 4629)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4629
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             572..1318
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKIINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     polyA_signal    1373..1849
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1572..1591)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   1889..2029
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1889..2018
                     /label=AAV2 ITR
     rep_origin      2104..2559
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2191..2210)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2401..2422
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2576..2595)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2695..2717
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2755..2773)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2841..2945
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2946..3806
                     /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"
     primer_bind     complement(3164..3183)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3977..4565
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4466..4485
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4627..127
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt
       61 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
      121 ggttcctgcg gccgcacgcg ccctgcagag ggccctgcgt atgagtgcaa gtgggtttta
      181 ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca
      241 agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca
      301 ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg ccttcgcccc
      361 cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg tcactcgccg
      421 gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag
      481 ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg
      541 gcggacggta tcgataagct tacccacaac catgtccaga ctggacaaga gcaaaatcat
      601 aaacagcgct ctggaattac tcaatggagt cggtatcgaa ggcctgacga caaggaaact
      661 cgctcaaaag ctgggagttg agcagcctac cctgtactgg cacgtgaaga acaagcgggc
      721 cctgctcgat gccctgccaa tcgagatgct ggacaggcat catacccaca gctgccccct
      781 ggaaggcgag tcatggcaag actttctgcg gaacaacgcc aagtcatacc gctgtgctct
      841 cctctcacat cgcgacgggg ctaaagtgca tctcggcacc cgcccaacag agaaacagta
      901 cgaaaccctg gaaaatcagc tcgcgttcct gtgtcagcaa ggcttctccc tggagaacgc
      961 actgtacgct ctgtccgccg tgggccactt tacactgggc tgcgtattgg aggaacagga
     1021 gcatcaagta gcaaaagagg aaagagagac acctaccacc gattctatgc ccccacttct
     1081 gaagcaagca attgagctgt tcgaccggca gggagccgaa cctgccttcc ttttcggcct
     1141 ggaactaatc atatgtggcc tggagaaaca gctaaagtgc gaaagcggcg ggccgaccga
     1201 cgcccttgac gattttgact tagacatgct cccagccgat gcccttgacg actttgacct
     1261 tgatatgctg cctgctgacg ctcttgacga ttttgacctt gacatgctcc ccgggtaagc
     1321 tagagtcgac ctgcagaagc ttgcctcgag cagcgctgct cgagagatct acgggtggca
     1381 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac
     1441 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa
     1501 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag
     1561 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc
     1621 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt
     1681 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac
     1741 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc
     1801 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta
     1861 ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     1921 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     1981 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     2041 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     2101 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2161 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2221 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2281 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     2341 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2401 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     2461 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     2521 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     2581 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     2641 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     2701 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     2761 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     2821 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2881 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2941 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3001 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3061 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3121 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3181 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3241 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3301 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3361 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3421 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3481 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3541 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3601 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3661 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3721 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3781 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3841 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3901 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3961 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4021 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4081 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4141 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4201 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4261 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4321 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4381 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4441 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4501 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4561 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4621 acatgtcct
//
