LOCUS       sequence-117710-        7719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flp-dependent expression of rabies glycoprotein to mediate 
            trans-synaptic spread of modifed rabies virus in cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7719)
  AUTHORS   Schwarz LA, Miyamichi K, Gao XJ, Beier KT, Weissbourd B, DeLoach KE,
            Ren J, Ibanes S, Malenka RC, Kremer EJ, Luo L
  TITLE     Viral-genetic tracing of the input-output organization of a central 
            noradrenaline circuit.
  JOURNAL   Nature. 2015 Aug 6;524(7563):88-92. doi: 10.1038/nature14600. Epub 
            2015 Jul 1.
  PUBMED    26131933
REFERENCE   2  (bases 1 to 7719)
  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: 
            "10.1038/nature14600"; journalName: "Nature"; date: "2015-08-6- 6"; 
            volume: "524"; issue: "7563"; pages: "88-92"
COMMENT     Sequence Label: CAG-FLEx(FRT)-G
FEATURES             Location/Qualifiers
     source          1..7719
                     /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"
     repeat_region   12..141
                     /label=AAV2 ITR
     enhancer        227..530
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        532..809
                     /label=chicken beta-actin promoter
     intron          811..1828
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1752..1774
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1836..1855
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1910..1957
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      3644..3653
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3659..3706)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    3848..4436
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3901..3921)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4319..4330)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4460..4936
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4659..4678)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4982..5122
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4982..5111
                     /label=AAV2 ITR
     rep_origin      5197..5652
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5284..5303)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5494..5515
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5669..5688)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5788..5810
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5848..5866)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5934..6038
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6039..6107,6108..6899)
                     /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(6257..6276)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7070..7658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7559..7578
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtctgca ggaatctagt tattaatagt
      181 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      241 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      301 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      361 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      421 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      481 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga
      541 gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt
      601 tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc
      661 caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggagaggt gcggcggcag
      721 ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc
      781 cctataaaaa gcgaagcgcg cggcgggcgg ggagtcgctg cgacgctgcc ttcgccccgt
      841 gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc
      901 acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct tggtttaatg
      961 acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg agggcccttt
     1021 gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg
     1081 cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc tttgtgcgct
     1141 ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg gggggggctg
     1201 cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg
     1261 cgcgtcggtc gggctgcaac cccccctgca cccccctccc cgagttgctg agcacggccc
     1321 ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg ctcgccgtgc cgggcggggg
     1381 gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg agggctcggg
     1441 ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca
     1501 ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc aaatctgtgc
     1561 ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg cgaagcggtg
     1621 cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc gccgccgtcc
     1681 ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc gggggggacg
     1741 gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca
     1801 tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc
     1861 tcatcatttt ggcaaagaat tgatttgata ccgcgggccc gggggtaccg aagttcctat
     1921 tccgaagttc ctattctcta gaaagtatag gaacttcatc aaaataggaa gaccaatgct
     1981 tcaccatcga cccgaattgc caagcatcac catcgacaga agttcctatt ccgaagttcc
     2041 tattcttcaa aaggtatagg aacttcgtcg acttacagtc tggtctcacc cccactcttg
     2101 tgtgattccc atgaagatat gatcttcccg ctttggggag tgactgacac ctccctccct
     2161 gtccctctga gattgtgttg cgtaggttct gatcgattga ctcttctaca acatgtcatc
     2221 aggaaaatta tcaacatcaa ggcagtcagg gcccctgcac tcagtaatac atacttcccc
     2281 cagttcggga gacccaagtc aactcctgag acctgattgt gcacatcggg aaggtgaact
     2341 tcaacaaaat cctcagcctc gtcaccgtcc ttgaaaacgg tagacgggtc tgccaggggg
     2401 tgcacaaggg ggataaccga ggattccaac aactccatat gttgctggag gagggatgat
     2461 tgcatctctg ggattaagac attgccgtca ggtcctaata ttataccatt gaaaaacacc
     2521 ccgttcacat gaggatgaca cctcccccca actcttaaac acccttttga agggaggatc
     2581 tcattccaag ttctgactga cttgtagtga gcatcggctt ccatcaaggt cttgttgaat
     2641 atggtatatg cttttccaaa cccagggaca agttttctta aatgactgag acgtctgaaa
     2701 ctcactgact tggttgtcat gatggactct agtgcatcca gacactcctc tctcttcctg
     2761 accaactcct ctacaacaag gtgctcaatt tcgtctgagc gaaagtcgtg caggttcacc
     2821 aacttatcgg gagggcacca tttggtttca tttgatgttt gcatcgagac ccatgttcca
     2881 tccataagtc taagtcctag aactccacat aacttgagtt tgcatgctcc ttttaaagac
     2941 ttatataggc ctctttcatc tacaaagccg caagtctcac tccctttgga tgctctcttc
     3001 cctctactat tggtaaaaat gtcacaagac atccctagtc tcggattctc gggcatccaa
     3061 atggtgtaat cgtggttagt ggagcagtag gtagaagaca ccgctactcc tgagcacttc
     3121 ccgctaggga agaccctcga gtgaagggat ctgtcatatg ggtccaaatc tgccacactt
     3181 ggagatatga taacgagaga ctccttggtg gtttttacag ttcgaagcca gcggtagtca
     3241 gggtacggat tgtgtagaga ctcttcatat ctggggtcac cggccatctt ccagttgtac
     3301 gcggctctac atgcatctgg tgttgggcgg aaatgctttc ttttgaacgt ggttgtgaca
     3361 taaccaacga agttagtgta ggtttcagcc tccgtcacaa cgcctgtgca agtgaacccg
     3421 ttcactttta tggctaagat gtatccaact ttaagttcca tgtaggagaa ccctgacagg
     3481 ttggtgcatc cttcgtcctc cactaccaaa ttgtttgggc agctgaggtg atgtatgtca
     3541 atcggactcc agggaccaag cttgtctggt atcgtgtaaa tagggaattt cccaaaacac
     3601 aatggaaaaa ccagaagggg tacaaacagg agagcctgag gaaccatggt ggcgcgccga
     3661 agttcctata ctttctagag aataggaact tcggaatagg aacttccgtt gggattcttc
     3721 ctattttgat ccaagcatca ccatcgaccc tctagtccag atctcaccat cgacccgaag
     3781 ttcctatacc ttttgaagaa taggaacttc ggaataggaa cttcaagctt atatgcggcc
     3841 gatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3901 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3961 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4021 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4081 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4141 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4201 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     4261 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4321 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4381 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     4441 ataccgtaga tcgatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     4501 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     4561 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     4621 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     4681 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     4741 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     4801 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     4861 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     4921 tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgaact agtgcggccg
     4981 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     5041 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     5101 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     5161 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     5221 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     5281 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5341 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5401 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5461 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5521 cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct
     5581 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     5641 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5701 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     5761 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     5821 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     5881 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5941 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     6001 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     6061 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     6121 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     6181 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     6241 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     6301 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     6361 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6421 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     6481 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6541 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     6601 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     6661 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6721 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6781 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6841 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6901 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6961 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     7021 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     7081 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     7141 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     7201 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     7261 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     7321 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     7381 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     7441 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     7501 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     7561 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     7621 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     7681 tacggttcct ggccttttgc tggccttttg ctcacatgt
//
