LOCUS       sequence-377175-        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(r)
FEATURES             Location/Qualifiers
     source          1..5171
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   19..148
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    163..287
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        465..567
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             568..1251
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     complement(634..653)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1571..1876
                     /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     1755..1774
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2305..2429)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     repeat_region   2452..2592
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2452..2581
                     /label=AAV2 ITR
     rep_origin      2667..3122
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2754..2773)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2964..2985
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3139..3158)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3258..3280
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3318..3336)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3404..3508
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3509..3577,3578..4369)
                     /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(3727..3746)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4540..5128
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5029..5048
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccttttgc tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc
       61 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      121 gtggccaact ccatcactag gggttcctgc ggccgcacgc gtaccacttt gtacaagaaa
      181 gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa
      241 aaacagacta cataatactg taaaacacaa catatccagt cactatgaat caactactta
      301 gatggtatta gtgacctgta gtcgactaag ttggcagcat cacccgacgc actttgcgcc
      361 gaataaatac ctgtgacgga agatcacttc gcagaataaa taaatcctgg tgtccctgtt
      421 gataccggga agccctgggc caacttttgg cgaaaatgag acgttgatcg gcacgtaaga
      481 ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg
      541 agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt
      601 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
      661 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
      721 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg
      781 gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
      841 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
      901 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
      961 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg
     1021 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
     1081 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
     1141 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
     1201 tgcgatgagt ggcagggggg gcgtaaacgc cgcgtggatc cggcttacta aaagccagat
     1261 aacagtatgc gtatttgcgc gctgattttt gcggtataag aatatatact gatatgtata
     1321 cccgaagtat gtcaaaaaga ggtatgctat gaagcagcgt attacagtga cagttgacag
     1381 cgacagctat cagttgctca aggcatatat gatgtcaata tctccggtct ggtaagcaca
     1441 accatgcaga atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa
     1501 gggatggctg aggtcgcccg gtttattgaa atgaacggct cttttgctga cgagaacagg
     1561 ggctggtgaa atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt
     1621 tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc
     1681 cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg
     1741 ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg
     1801 ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat
     1861 gttctgggga atataaatgt caggctccct tatacacagc cagtctgcag gtcgatacag
     1921 tagaaattac agaaacttta tcacgtttag taagtataga ggctgaaaat ccagatgaag
     1981 ccgaacgact tgtaagagaa aagtataaga gttgtgaaat tgttcttgat gcagatgatt
     2041 ttcaggacta tgacactagc gtatatgaat aggtagatgt ttttattttg tcacacaaaa
     2101 aagaggctcg cacctctttt tcttatttct ttttatgatt taatacggca ttgaggacaa
     2161 tagcgagtag gctggatacg acgattccgt ttgagaagaa catttggaag gctgtcggtc
     2221 gactaagttg gcagcatcac ccgaagaaca tttggaaggc tgtcggtcga ctacaggtca
     2281 ctaataccat ctaagtagtt gattcatagt gactggatat gttgtgtttt acagtattat
     2341 gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt
     2401 tctcgttcag cttttttgta caaacttgtc acgtgcggac cgagcggccg caggaacccc
     2461 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     2521 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     2581 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     2641 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     2701 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
     2761 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2821 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     2881 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     2941 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
     3001 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
     3061 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     3121 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     3181 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     3241 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     3301 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     3361 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3421 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3481 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3541 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3601 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3661 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3721 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3781 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3841 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3901 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3961 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4021 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4081 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4141 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4201 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4261 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4321 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4381 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4441 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4501 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4561 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4621 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4681 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4741 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4801 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4861 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4921 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4981 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5041 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5101 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5161 ggccttttgc t
//
