LOCUS       sequence-139908-        6562 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pAAV vector to express oG (optimized Glycoprotein) in a 
            Flp-dependent manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6562)
  AUTHORS   Kim EJ, Jacobs MW, Ito-Cole T, Callaway EM
  TITLE     Improved Monosynaptic Neural Circuit Tracing Using Engineered Rabies
            Virus Glycoproteins.
  JOURNAL   Cell Rep. 2016 Apr 26;15(4):692-699. doi: 
            10.1016/j.celrep.2016.03.067. Epub 2016 Apr 14.
  PUBMED    27149846
REFERENCE   2  (bases 1 to 6562)
  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"; doi: 
            "10.1016/j.celrep.2016.03.067"; journalName: "Cell Rep"; date: 
            "2016-04-26- 26"; volume: "15"; issue: "4"; pages: "692-699"
COMMENT     Sequence Label: pAAV-CAG-fDIO-oG-WPRE-SV40pA
FEATURES             Location/Qualifiers
     source          1..6562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2069..2090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2281..2300
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2459..2588
                     /label=AAV2 ITR
     promoter        2901..3178
                     /label=chicken beta-actin promoter
     primer_bind     3458..3480
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3542..3561
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    3597..3630
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     regulatory      5294..5303
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(5319..5352)
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     misc_feature    5464..6052
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5517..5537)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5935..5946)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6095..6216
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6106..6125)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6160..6179
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   6378..6507
                     /label=AAV2 ITR
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag
     2641 gaagatcgta ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     2701 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     2761 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     2821 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     2881 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
     2941 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
     3001 gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg
     3061 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
     3121 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
     3181 agtcgctgcg acgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
     3241 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
     3301 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
     3361 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ctgtccgcgg
     3421 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     3481 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     3541 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattgga tcctccgaag
     3601 ttcctattct tcaaatagta taggaacttc gcagaatggt agctggattg tagctgctat
     3661 tagcaatatg aaacctctta gaagttccta ttcttcaaaa ggtataggaa cttcggcgcg
     3721 ccttagagcc gtgtctcgcc cccagattta tgactttccc aactgctgat gattttaccg
     3781 ctctggggtg tgacagacac ttctctaccg gtccccctca gattgtgctg agttggctcg
     3841 gagcggttca ctcgtcgcca gcaggtcatc aggaaaatga tcagcatgag agcagtcagt
     3901 gctccagcgg agagcagcac gtacttgccc caattaggca gtccgagatc gacgccactg
     3961 atccgttcgt gcacgtcggg caggtggacc tccacaaaat cttcggcctc gtcgccgttt
     4021 ttgaacactg tgcttgggtc tgcgagagga tgcatcaggg gaatgacgct tgacacgagc
     4081 agttccatgt gctgctggag cagactagac tgcatctcgg ggatcagcac gttaccatct
     4141 ggccccagaa tgataccatt aaagaagacc ccgttcacat gtgggtgaca tcgacccccg
     4201 actcgcaggc atcccttact aggaatgatt tcattccagg ttctcacaga tttataatgg
     4261 gcgtctgcct ccatcagtgt cttgttaaag atggtgtatg cttttccgaa gccagggacg
     4321 agcttcctca ggtggctcag cctcctgaag ctcacgctct tggtagtcat aattgactcc
     4381 agagcatcga gacattcctc ccttttcttc acgagttcct cgaccaccag atgttcgatc
     4441 tcatcgcttc ggaagtcgtg cagattcacg agctgcccag gggggcacca cttagtctcg
     4501 ttagatgtct gcatagcgac ccaggttccg tccatcagcc ggagtcccag cacgccacag
     4561 agtttcagct tgcaggcacc tttgaggctc ttatacagcc ctctttcatc cacaaatcca
     4621 caagtctcgc tgccttttga ggccctcttg cccctgctgt tggtgaagat gtcgcaggac
     4681 atccccagcc ttggattctc aggcatccag attgtataat catggttggt ggaacagtaa
     4741 gtggagctga cggccactcc ggagcagttt ccgccgggaa acactggact gtggagagat
     4801 cggtcgtagg ggtccaggtc ggcgacagat ggggaaatga tgaccaggct ctcttttgtg
     4861 gtcttcacag tgcgcagcca atgatagtct gggtaaggat tgtgcaggct ttcctcatag
     4921 cgggggtcgc cagccatctt ccagttgtaa gctgcccgac aggcgtctgg tgttggccga
     4981 aagtgttttc tcttgaaagt tgtggtgaca tatcccacaa aatttgtgta ggtttctgcc
     5041 tcggtgacca caccagtgca tgtgaacccg ttcatcttaa tagcagagat atagcccact
     5101 ttcagctcca tgtaggagaa tccgctcagg ttggtacagc cttcgtcctc gaccacgaga
     5161 ttgttagggc atgacagatg gtgaatatcg ataggactcc aggggcccag cttgtcgggg
     5221 atagtgtaga tagggaattt accaaagcag agtgggaaga cgagcagagg gacgaagagg
     5281 agagcctgtg ggaccatggt ggcggcgcta gcactagaga agttcctata ctatttgaag
     5341 aataggaact tcttgcctta acccagaaat tatcactgtt attctttaga atggtgcaaa
     5401 gagaagttcc tatacctttt gaagaatagg aacttcgaag aattcgatat caagcttatc
     5461 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     5521 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     5581 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     5641 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     5701 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     5761 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     5821 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     5881 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     5941 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     6001 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
     6061 cgtcgacccg ggcggccgct tcgagcagac atgataagat acattgatga gtttggacaa
     6121 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     6181 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     6241 atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     6301 tgtggtaaaa tcgataagga tcttcctaga gcatggctac gtagataagt agcatggcgg
     6361 gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     6421 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     6481 ggcctcagtg agcgagcgag cgcgcagctg cattaatgaa tcggccaacg cgcggggaga
     6541 ggcggtttgc gtattgggcg ct
//
