LOCUS       sequence-81274-e        8615 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct used for viral production of virus expressing the TVA 
            receptor, Rabies protein G and a marker protein TagBFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8615)
  AUTHORS   Betley JN, Cao ZF, Ritola KD, Sternson SM
  TITLE     Parallel, redundant circuit organization for homeostatic control of 
            feeding behavior.
  JOURNAL   Cell. 2013 Dec 5;155(6):1337-50. doi: 10.1016/j.cell.2013.11.002.
  PUBMED    24315102
REFERENCE   2  (bases 1 to 8615)
  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.cell.2013.11"; journalName: "Cell"; date: "2013-12-5- 5";
            volume: "155"; issue: "6"; pages: "1337-50"
COMMENT     Sequence Label: AAV-FLEX-rev-BFP-2a-TVA-2a-RabiesG
FEATURES             Location/Qualifiers
     source          1..8615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        96..399
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        401..678
                     /label=chicken beta-actin promoter
     intron          680..1697
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1621..1643
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1705..1724
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1801..1834)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(1845..1878)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             complement(3473..3532)
                     /codon_start=1
                     /product="2A peptide from equine rhinitis A virus 
                     polyprotein"
                     /label=E2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="QCTNYALLKLAGDVESNPGP"
     CDS             complement(4022..4075)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(4091..4114)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(4118..4141)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(4145..4168)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      4882..4891
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    4918..4951
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4962..4995
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     misc_feature    5019..5607
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5072..5092)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5490..5501)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5642..5659)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5648..5855
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5878..6018
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5878..6007
                     /label=AAV2 ITR
     rep_origin      6093..6548
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6180..6199)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6390..6411
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6565..6584)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6684..6706
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6744..6762)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6830..6934
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6935..7003,7004..7795)
                     /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(7153..7172)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7966..8554
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8455..8474
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcggccg cacgcgaaac aattctgcag gaatctagtt attaatagta atcaattacg
       61 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      121 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      181 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      241 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      301 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      361 tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc
      421 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt
      481 taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg
      541 gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc
      601 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag
      661 cgaagcgcgc ggcgggcggg gagtcgctgc gacgctgcct tcgccccgtg ccccgctccg
      721 ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg
      781 ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt
      841 cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga
      901 gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc
      961 tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc
     1021 gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca
     1081 aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg
     1141 ggctgcaacc ccccctgcac ccccctcccc gagttgctga gcacggcccg gcttcgggtg
     1201 cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg
     1261 tgggggtgcc gggcggggcg gggccgcctc gggccgggga gggctcgggg gaggggcgcg
     1321 gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat
     1381 ggtaatcgtg cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat
     1441 ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca
     1501 ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc
     1561 tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg
     1621 ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct
     1681 tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg
     1741 gcaaagaatt gatttgatac cgcgggcccc gcgtggacgg tacccttcag gtcgacgtcg
     1801 ataacttcgt atagcataca ttatacgaag ttatccgaat cgcaataact tcgtatagga
     1861 tactttatac gaagttattt ggcgcgccac tagttcacaa gcgcgtttca cctcctgact
     1921 tgtgtgactc ccacgacgaa atgatcttcc cggactgcgg agtgacggac acttctctcc
     1981 ccgtacccct caagttgtgc tgtgtgggtt ccgatctgtt cacccttcta cagcacgtca
     2041 tgaggaagat gatgagcatg agtgccgtca acgcgcctgc gctcaacagg acgtactttc
     2101 cccaattcgg caaaccgagg tccaccccgg acacttggtt atgtacgtcc gggaggtgga
     2161 cttccacaaa atcttccgct tcatctccgt ccttaaagac ggtggaaggg tcggccaacg
     2221 ggtgcacgag cggaattact gagctttcga gcaactccat gtgttgctga aggagggatg
     2281 actgcatttc gggaatcagg acattcccat cgggaccgag gataattcca ttgaaaaaca
     2341 ctccattgac atgggggtgg caccgtcctc cgactctcag acaacctttt gagggaagaa
     2401 tctcgttcca cgtccgtacc gacttatagt gcgcgtcggc ttccataagg gttttgttaa
     2461 agatggtgta tgcctttccg aacccaggga caagtttgcg gaggtggctc agccggcgaa
     2521 aggacaccga cttcgtagtc ataattgatt ccagcgcgtc cagacactct tccctttttc
     2581 gtacaagttc ctcgacgaca aggtgttcga tttcgtctga tctaaagtca tgaaggttca
     2641 cgagcttgtc gggtgggcac cacttcgttt cgttggacgt ctgcatcgac acccacgttc
     2701 catccataag cctaagtccg aggaccccac acagtttcag tttgcacgcc cccttgaggg
     2761 acttatacag acccctctca tcgacaaatc cgcacgtttc tgaccccttt gatgccctct
     2821 ttcctcttga gttcgtgaaa atgtcacatg acattccgag tctcggattc tccggcatcc
     2881 agatcgtata atcgtgattg gtcgaacaat acgtgctgct gacggctacc cccgagcatt
     2941 ttccgctagg aaacacgcgc gagtggaggg accgatcata cgggtccaaa tccgcgaccg
     3001 acggcgaaat gattacgagc gactctttcg ttgtttttac tgtcctcaac cagcggtaat
     3061 caggataggg gttatgcagt gattcttcgt agcgcgggtc cccagccatc ttccagttgt
     3121 atgcagcccg acaggcgtcc ggcgtggggc gaaaatgctt tcttttgaag gtagtagtca
     3181 catagcccac gaagttcgtg tatgtctccg cctccgtcac cacgccggta cacgtgaacc
     3241 cattgacttt gatggccagg atgtatccga ccttgagttc catgtacgag aaccccgaca
     3301 agttggtgca tccctcatcc tccactacga gattgttggg gcaggacaga tgatgaatat
     3361 cgatagggga ccagggtccc agtttatcag gaattgtgta gatagggaac tttccgaaac
     3421 acaggggaaa cacgagcaaa gggacaaaca aaagggcttg cgggactccg gaaggaccgg
     3481 ggttactttc aacatcgcca gcgagtttca acaaagcgta gttagtacat tgaccactac
     3541 cgctagcgga gaacaagtct gcctggctct tgtcaggcac cagcagctcc ttggatgcgc
     3601 tttcaagact gaagatgtca gacctgcttt ttgctttgga cttcccccat gcagcgatac
     3661 cacccacagc taccaggcag cacaggagca ctgcagtgat cagcatccac atgcggccgt
     3721 gattcctggc tggcagagca cgtccaggag caggggcagt gggagcctct gtgccgttgt
     3781 ccgtgggcac cgcggggatc gcgctggtcc cgcagcccca ctcgtcccgc ccgtcgtcgc
     3841 agtcggggtg tccgtcgcac agccagtctt gcgggtaaca ctccccgtgg gcaccgggcg
     3901 gctccgagca gcggaactga ccgggggggc aacgggacaa agaaccgtta ccggacccgt
     3961 taccggtcac gttaccgggc agcagcagca gcagcagcgc gggcagcagc cgcgcatcga
     4021 ttgggccagg attctcctcg acgtcaccgc atgttagcag acttcctctg ccctctccac
     4081 taccaagctt cttatcatca tcgtctttgt agtccccctt gtcatcatca tccttgtagt
     4141 cgcctttgtc gtcgtcgtcc ttgtaatccc caccgttgag tttgtggccg agcttggagg
     4201 gaaggtcaca ataccgagcc accgcgactt cgtgctgctc gacgtaagtc tcgttgttcg
     4261 cctctttgat gcgttcaagt cggtagtcga catagtacac tccgggcatc ttgagattct
     4321 tggcaggctt tttgctccta tatgtggtct tggcgttcgc aataagatgc gaacccccta
     4381 ccaatttgag cgccatatca ttcctccctt ccagaccacc gtcagccggg tacagggttt
     4441 cagtgaatgc ctcccatccg agtgtctttt tctgcatgac cggaccattg gatgtgaaat
     4501 tgactcctct gatcttcacg ttgtagataa ggcacccgtc ttggagcgac gtgtcttgtg
     4561 tcgcagtcag cactccacca tcctcatacg tcgtcactct ctcccacgta aacccttcgg
     4621 gaaatgactg cttaaagaaa tcaggaatcc cctgcgtgtg attgataaac gttttcgatc
     4681 cataaaggaa cgaggttgcc agaatgtcga aggcaaaggg caatgggcct ccttccacta
     4741 ctttgatccg catggtctgt gtcccctcgt acggctttcc ctccccttct gacgtacact
     4801 tgaaatgatg gttatccact gttccctcca tatacagttt catatgcatg ttttccttga
     4861 tcaactcctc tcctttgctg accatggtgg cggcactagt ggatccggcc ggccttgata
     4921 acttcgtata atgtatgcta tacgaagtta tccgaatcgc aataacttcg tataaagtat
     4981 cctatacgaa gttatcgaag atcatcaagc ttatcgataa tcaacctctg gattacaaaa
     5041 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     5101 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     5161 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     5221 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     5281 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     5341 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     5401 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc
     5461 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     5521 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     5581 ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatcgatct
     5641 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     5701 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     5761 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     5821 gaggattggg aagacaatag caggcatgct ggggacacgt gcggaccgag cggccgcagg
     5881 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     5941 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     6001 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     6061 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     6121 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     6181 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     6241 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     6301 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     6361 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     6421 caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     6481 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     6541 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     6601 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     6661 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     6721 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     6781 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     6841 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     6901 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     6961 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     7021 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     7081 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     7141 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     7201 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     7261 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     7321 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     7381 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     7441 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     7501 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     7561 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     7621 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     7681 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     7741 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     7801 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     7861 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     7921 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     7981 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     8041 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     8101 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     8161 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     8221 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     8281 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     8341 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     8401 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     8461 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     8521 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     8581 gttcctggcc ttttgctggc cttttgctca catgt
//
