LOCUS       sequence-376640-        5171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway destination vector, empty AAV vector backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5171)
  AUTHORS   Sohn J, Takahashi M, Okamoto S, Ishida Y, Furuta T, Hioki H
  TITLE     A Single Vector Platform for High-Level Gene Transduction of Central
            Neurons: Adeno-Associated Virus Vector Equipped with the Tet-Off 
            System.
  JOURNAL   PLoS One. 2017 Jan 6;12(1):e0169611. doi: 
            10.1371/journal.pone.0169611. eCollection 2017.
  PUBMED    28060929
REFERENCE   2  (bases 1 to 5171)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2017-01-6"; volume: "12(1):e0169611. doi"; pages: " 
            10.1371/journal.pone.0169611. eCollection 2017"
COMMENT     Sequence Label: pAAV2-DEST(f)
FEATURES             Location/Qualifiers
     source          1..5171
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   5..134
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    149..273
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(702..1007)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(804..823)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1327..2010)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     1925..1944
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2011..2113)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(2291..2415)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     repeat_region   2438..2578
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2438..2567
                     /label=AAV2 ITR
     rep_origin      2653..3108
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2740..2759)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2950..2971
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3125..3144)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3244..3266
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3304..3322)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3390..3494
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3495..3563,3564..4355)
                     /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(3713..3732)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4526..5114
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5015..5034
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac
       61 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
      121 cactaggggt tcctgcggcc gcacgcgtac aagtttgtac aaaaaagctg aacgagaaac
      181 gtaaaatgat ataaatatca atatattaaa ttagattttg cataaaaaac agactacata
      241 atactgtaaa acacaacata tccagtcact atgaatcaac tacttagatg gtattagtga
      301 cctgtagtcg accgacagcc ttccaaatgt tcttcgggtg atgctgccaa cttagtcgac
      361 cgacagcctt ccaaatgttc ttctcaaacg gaatcgtcgt atccagccta ctcgctattg
      421 tcctcaatgc cgtattaaat cataaaaaga aataagaaaa agaggtgcga gcctcttttt
      481 tgtgtgacaa aataaaaaca tctacctatt catatacgct agtgtcatag tcctgaaaat
      541 catctgcatc aagaacaatt tcacaactct tatacttttc tcttacaagt cgttcggctt
      601 catctggatt ttcagcctct atacttacta aacgtgataa agtttctgta atttctactg
      661 tatcgacctg cagactggct gtgtataagg gagcctgaca tttatattcc ccagaacatc
      721 aggttaatgg cgtttttgat gtcattttcg cggtggctga gatcagccac ttcttccccg
      781 ataacggaga ccggcacact ggccatatcg gtggtcatca tgcgccagct ttcatccccg
      841 atatgcacca ccgggtaaag ttcacgggag actttatctg acagcagacg tgcactggcc
      901 agggggatca ccatccgtcg cccgggcgtg tcaataatat cactctgtac atccacaaac
      961 agacgataac ggctctctct tttataggtg taaaccttaa actgcatttc accagcccct
     1021 gttctcgtca gcaaaagagc cgttcatttc aataaaccgg gcgacctcag ccatcccttc
     1081 ctgattttcc gctttccagc gttcggcacg cagacgacgg gcttcattct gcatggttgt
     1141 gcttaccaga ccggagatat tgacatcata tatgccttga gcaactgata gctgtcgctg
     1201 tcaactgtca ctgtaatacg ctgcttcata gcatacctct ttttgacata cttcgggtat
     1261 acatatcagt atatattctt ataccgcaaa aatcagcgcg caaatacgca tactgttatc
     1321 tggcttttag taagccggat ccacgcggcg tttacgcccc ccctgccact catcgcagta
     1381 ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac
     1441 ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa
     1501 aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac
     1561 ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag
     1621 gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc
     1681 gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca
     1741 agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac ggaattccgg
     1801 atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt
     1861 tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca
     1921 ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac
     1981 ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga
     2041 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     2101 tacgtgccga tcaacgtctc attttcgcca aaagttggcc cagggcttcc cggtatcaac
     2161 agggacacca ggatttattt attctgcgaa gtgatcttcc gtcacaggta tttattcggc
     2221 gcaaagtgcg tcgggtgatg ctgccaactt agtcgactac aggtcactaa taccatctaa
     2281 gtagttgatt catagtgact ggatatgttg tgttttacag tattatgtag tctgtttttt
     2341 atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc gttcagcttt
     2401 cttgtacaaa gtggtcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     2461 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     2521 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     2581 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     2641 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2701 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     2761 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2821 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     2881 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2941 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     3001 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     3061 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     3121 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     3181 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     3241 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     3301 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3361 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3421 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3481 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3541 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3601 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3661 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3721 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3781 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3841 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3901 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3961 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4021 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4081 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4141 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4201 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4261 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4321 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4381 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4441 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4501 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4561 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4621 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4681 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4741 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4801 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4861 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4921 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4981 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5041 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5101 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5161 cttttgctca c
//
