LOCUS       62056_2495        6513 bp DNA     circular SYN 30-NOV-2021
DEFINITION  Cloning vector pAAV-Syn-NS1-p2A-NLSdTomato, complete sequence.
ACCESSION   MZ695807
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6513)
  AUTHORS   Li E, Guo J, Oh SJ, Luo Y, Oliveros HC, Du W, Arano R, Kim Y, Chen 
            YT, Eitson J, Lin DT, Li Y, Roberts T, Schoggins JW, Xu W.
  TITLE     Anterograde transneuronal tracing and genetic control with 
            engineered yellow fever vaccine YFV-17D
  JOURNAL   Nat Methods (2021) In press
  PUBMED    34824475
REFERENCE   2  (bases 1 to 6513)
  AUTHORS   Xu W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-AUG-2021) Neuroscience, UT Southwestern Medical 
            Center, Dallas, TX 75229, USA
REFERENCE   3  (bases 1 to 6513)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Methods
            (2021) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-AUG-2021) Neuroscience, UT Southwestern Medical Center, Dallas, 
            TX 75229, USA"
FEATURES             Location/Qualifiers
     source          1..6513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     promoter        194..641
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             1871..1891
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             1889..2590
                     /codon_start=1
                     /label=dTomato
                     /note="dimeric variant of DsRed fluorescent protein (Shaner
                     et al., 2004)"
                     /translation="VVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     misc_feature    2640..3228
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    3260..3736
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   3776..3916
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      3991..4446
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4728..4832
                     /label=AmpR promoter
     CDS             4833..5690
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5864..6452
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtttaat taagtgtcta gactgcagag
      181 ggccctgcgt atgagtgcaa gtgggtttta ggaccaggat gaggcggggt gggggtgcct
      241 acctgacgac cgaccccgac ccactggaca agcacccaac ccccattccc caaattgcgc
      301 atcccctatc agagaggggg aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag
      361 cttcagcacc gcggacagtg ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca
      421 gcactgaagg cgcgctgacg tcactcgccg gtcccccgca aactcccctt cccggccacc
      481 ttggtcgcgt ccgcgccgcc gccggcccag ccggaccgca ccacgcgagg cgcgagatag
      541 gggggcacgg gcgcgaccat ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg
      601 cggtgggcag cggaggagtc gtgtcgtgcc tgagagcgca gtcgagaaac cggctagagg
      661 atccgccacc atgaacatga caatgtccat gagcatgatc ttggtaggag tgatcatgat
      721 gtttttgtct ctaggagttg gggcggatca aggatgcgcc atcaactttg gcaagagaga
      781 gctcaagtgc ggagatggta tcttcatatt tagagactct gatgactggc tgaacaagta
      841 ctcatactat ccagaagatc ctgtgaagct tgcatcaata gtgaaagcct cttttgaaga
      901 agggaagtgt ggcctaaatt cagttgactc ccttgagcat gagatgtgga gaagcagggc
      961 agatgagatc aatgccattt ttgaggaaaa cgaggtggac atttctgttg tcgtgcagga
     1021 tccaaagaat gtttaccaga gaggaactca tccattttcc agaattcggg atggtctgca
     1081 gtatggttgg aagacttggg gtaagaacct tgtgttctcc ccagggagga agaatggaag
     1141 cttcatcata gatggaaagt ccaggaaaga atgcccgttt tcaaaccggg tctggaattc
     1201 tttccagata gaggagtttg ggacgggagt gttcaccaca cgcgtgtaca tggacgcagt
     1261 ctttgaatac accatagact gcgatggatc tatcttgggt gcagcggtga acggaaaaaa
     1321 gagtgcccat ggctctccaa cattttggat gggaagtcat gaagtaaatg ggacatggat
     1381 gatccacacc ttggaggcat tagattacaa ggagtgtgag tggccactga cacatacgat
     1441 tggaacatca gttgaagaga gtgaaatgtt catgccgaga tcaatcggag gcccagttag
     1501 ctctcacaat catatccctg gatacaaggt tcagacgaac ggaccttgga tgcaggtacc
     1561 actagaagtg aagagagaag cttgcccagg gactagcgtg atcattgatg gcaactgtga
     1621 tggacgggga aaatcaacca gatccaccac ggatagcggg aaagttattc ctgaatggtg
     1681 ttgccgctcc tgcacaatgc cgcctgtgag cttccatggt agtgatgggt gttggtatcc
     1741 catggaaatt aggccaagga aaacgcatga aagccatctg gtgcgctcct gggttacagc
     1801 tgccactaac ttctccctgt tgaaacaagc aggggatgtc gaagagaatc ccgggccatc
     1861 tagaatggcc cccaagaaga agaggaaggt ggtgagcaag ggcgaggagg tcatcaaaga
     1921 gttcatgcgc ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt tcgagatcga
     1981 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     2041 gggcggcccc ctgcccttcg cctgggacat cctgtccccc cagttcatgt acggctccaa
     2101 ggcgtacgtg aagcaccccg ccgacatccc cgattacaag aagctgtcct tccccgaggg
     2161 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg tgacccagga
     2221 ctcctccctg caggacggca cgctgatcta caaggtgaag atgcgcggca ccaacttccc
     2281 ccccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct
     2341 gtacccccgc gacggcgtgc tgaagggcga gatccaccag gccctgaagc tgaaggacgg
     2401 cggccactac ctggtggagt tcaagaccat ctacatggcc aagaagcccg tgcaactgcc
     2461 cggctactac tacgtggaca ccaagctgga catcacctcc cacaacgagg actacaccat
     2521 cgtggaacag tacgagcgct ccgagggccg ccaccacctg ttcctgtacg gcatggacga
     2581 gctgtacaag taagtcgacg gcgcgcccct gcagggaatt cgatatcaag cttatcgata
     2641 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     2701 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     2761 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     2821 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     2881 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     2941 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3001 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     3061 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3121 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3181 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag
     3241 cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     3301 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     3361 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     3421 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     3481 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     3541 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     3601 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     3661 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     3721 tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa
     3781 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     3841 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     3901 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3961 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     4021 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4081 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4141 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4201 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4261 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4321 accctatctc gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4381 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     4441 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     4501 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4561 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4621 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     4681 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4741 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4801 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4861 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4921 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4981 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5041 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5101 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5161 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5221 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5281 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5341 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5401 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5461 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5521 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5581 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5641 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5701 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5761 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5821 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5881 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5941 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6001 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6061 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6121 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6181 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6241 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6301 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6361 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6421 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6481 tcctggcctt ttgctggcct tttgctcaca tgt
//
