LOCUS       sequence-127049-        7277 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flp-dependent expression of TVA-mCherry fusion to mediate modifed 
            rabies virus expression in cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7277)
  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 7277)
  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)-TC
FEATURES             Location/Qualifiers
     source          1..7277
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        216..519
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        521..798
                     /label=chicken beta-actin promoter
     intron          799..1815
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1739..1761
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1823..1842
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1897..1944
                     /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)."
     CDS             complement(2060..2767)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2338..2357)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2607..2625
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3202..3211
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3217..3264)
                     /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    3406..3994
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3459..3479)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3877..3888)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4018..4494
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4217..4236)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4540..4680
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4540..4669
                     /label=AAV2 ITR
     rep_origin      4755..5210
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4842..4861)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5052..5073
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5227..5246)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5346..5368
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5406..5424)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5492..5596
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5597..5665,5666..6457)
                     /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(5815..5834)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6628..7216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7117..7136
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtctgcag gaatctagtt attaatagta atcaattacg
      181 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      241 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      301 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      361 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      421 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      481 tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc
      541 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt
      601 taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg
      661 gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc
      721 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag
      781 cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc
      841 gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg
      901 cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct
      961 tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc
     1021 ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg
     1081 cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc
     1141 gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa
     1201 ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg
     1261 ctgcaacccc ccctgcaccc ccctccccga gttgctgagc acggcccggc ttcgggtgcg
     1321 gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg
     1381 ggggtgccgg gcggggcggg gccgcctcgg gccggggagg gctcggggga ggggcgcggc
     1441 ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg
     1501 taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct
     1561 gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg cgccggcagg
     1621 aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc
     1681 cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg
     1741 ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt tcatgccttc
     1801 ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca tcattttggc
     1861 aaagaattga tttgataccg cgggcccggg ggtaccgaag ttcctattcc gaagttccta
     1921 ttctctagaa agtataggaa cttcatcaaa ataggaagac caatgcttca ccatcgaccc
     1981 gaattgccaa gcatcaccat cgacagaagt tcctattccg aagttcctat tcttcaaaag
     2041 gtataggaac ttcgtcgact tacttgtaca gctcgtccat gccgccggtg gagtggcggc
     2101 cctcggcgcg ttcgtactgt tccacgatgg tgtagtcctc gttgtgggag gtgatgtcca
     2161 acttgatgtt gacgttgtag gcgccgggca gctgcacggg cttcttggcc ttgtaggtgg
     2221 tcttgacctc agcgtcgtag tggccgccgt ccttcagctt cagcctctgc ttgatctcgc
     2281 ccttcagggc gccgtcctcg gggtacatcc gctcggagga ggcctcccag cccatggtct
     2341 tcttctgcat tacggggccg tcggagggga agttggtgcc gcgcagcttc accttgtaga
     2401 tgaactcgcc gtcctgcagg gaggagtcct gggtcacggt caccacgccg ccgtcctcga
     2461 agttcatcac gcgctcccac ttgaagccct cggggaagga cagcttcaag tagtcgggga
     2521 tgtcggcggg gtgcttcacg taggccttgg agccgtacat gaactgaggg gacaggatgt
     2581 cccaggcgaa gggcaggggg ccacccttgg tcaccttcag cttggcggtc tgggtgccct
     2641 cgtaggggcg gccctcgccc tcgccctcga tctcgaactc gtggccgttc acggagccct
     2701 ccatgtgcac cttgaagcgc atgaactcct tgatgatggc catgttatcc tcctcgccct
     2761 tgctcacccc accccccttg gatgcgcttt caagactgaa gatgtcagac ctgctttttg
     2821 ctttggactt cccccatgca gcgataccac ccacagctac caggcagcac aggagcactg
     2881 cagtgatcag catccacatg cggccgtgat tcctggctgg cagagcacgt ccaggagcag
     2941 gggcagtggg agcctctgtg ccgttgtccg tgggcaccgc ggggatcgcg ctggtcccgc
     3001 agccccactc gtcccgcccg tcgtcgcagt cggggtgtcc gtcgcacagc cagtcctgcg
     3061 ggtaacactc cccgtgggca ccgggcggct ccgagcagcg gaactgaccg ggggggcaac
     3121 gggacaaaga accgttaccg gacccgttac cggtcacgtt accgggcagc agcagcagca
     3181 gcagcgcggg cagcagccgc gccatggtgg cgcgccgaag ttcctatact ttctagagaa
     3241 taggaacttc ggaataggaa cttccattgg gattcttcct attttgatcc aagcatcacc
     3301 atcgaccctc tagtccagat ctcaccatcg acccgaagtt cctatacctt ttgaagaata
     3361 ggaacttcgg aataggaact tcaagcttat atgcggccga tcgataatca acctctggat
     3421 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     3481 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     3541 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     3601 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     3661 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     3721 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3781 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     3841 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     3901 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     3961 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtagatc gatctacggg
     4021 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
     4081 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
     4141 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
     4201 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
     4261 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
     4321 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
     4381 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
     4441 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
     4501 ttgtaggtaa ccacgtgcgg accgaactag tgcggccgca ggaaccccta gtgatggagt
     4561 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     4621 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     4681 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     4741 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4801 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     4861 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4921 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     4981 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     5041 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     5101 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     5161 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     5221 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     5281 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     5341 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     5401 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     5461 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     5521 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5581 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5641 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5701 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5761 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5821 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5881 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5941 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     6001 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     6061 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     6121 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     6181 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     6241 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     6301 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     6361 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6421 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     6481 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6541 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6601 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6661 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6721 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6781 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6841 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6901 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6961 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     7021 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     7081 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     7141 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     7201 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     7261 gccttttgct cacatgt
//
