LOCUS       sequence-208797-        8709 bp    DNA     linear   UNA 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      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8709)
  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 8709)
  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..8709
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          134..1150
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1074..1096
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1158..1177
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1254..1287)
                     /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(1298..1331)
                     /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(2926..2985)
                     /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(3473..3526)
                     /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(3542..3565)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(3569..3592)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(3596..3619)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      4333..4342
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    4369..4402
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4413..4446
                     /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    4470..5058
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4523..4543)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4941..4952)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5093..5110)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5099..5306
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5329..5469
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5329..5458
                     /label=AAV2 ITR
     rep_origin      5544..5999
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5631..5650)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5841..5862
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6016..6035)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6135..6157
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6195..6213)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6281..6385
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6386..6454,6455..7246)
                     /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(6604..6623)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7417..8005
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7906..7925
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   8067..8196
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        8289..8592
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
       61 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
      121 cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc
      181 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
      241 cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct
      301 gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc
      361 ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg
      421 cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg
      481 gagcgcggcc gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg
      541 cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca
      601 accccccctg cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct
      661 ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt
      721 gccgggcggg gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc
      781 ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc
      841 gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag
      901 gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga
      961 aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc
     1021 tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg
     1081 cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt
     1141 tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga
     1201 attgatttga taccgcgggc cccgcgtgga cggtaccctt caggtcgacg tcgataactt
     1261 cgtatagcat acattatacg aagttatccg aatcgcaata acttcgtata ggatacttta
     1321 tacgaagtta tttggcgcgc cactagttca caagcgcgtt tcacctcctg acttgtgtga
     1381 ctcccacgac gaaatgatct tcccggactg cggagtgacg gacacttctc tccccgtacc
     1441 cctcaagttg tgctgtgtgg gttccgatct gttcaccctt ctacagcacg tcatgaggaa
     1501 gatgatgagc atgagtgccg tcaacgcgcc tgcgctcaac aggacgtact ttccccaatt
     1561 cggcaaaccg aggtccaccc cggacacttg gttatgtacg tccgggaggt ggacttccac
     1621 aaaatcttcc gcttcatctc cgtccttaaa gacggtggaa gggtcggcca acgggtgcac
     1681 gagcggaatt actgagcttt cgagcaactc catgtgttgc tgaaggaggg atgactgcat
     1741 ttcgggaatc aggacattcc catcgggacc gaggataatt ccattgaaaa acactccatt
     1801 gacatggggg tggcaccgtc ctccgactct cagacaacct tttgagggaa gaatctcgtt
     1861 ccacgtccgt accgacttat agtgcgcgtc ggcttccata agggttttgt taaagatggt
     1921 gtatgccttt ccgaacccag ggacaagttt gcggaggtgg ctcagccggc gaaaggacac
     1981 cgacttcgta gtcataattg attccagcgc gtccagacac tcttcccttt ttcgtacaag
     2041 ttcctcgacg acaaggtgtt cgatttcgtc tgatctaaag tcatgaaggt tcacgagctt
     2101 gtcgggtggg caccacttcg tttcgttgga cgtctgcatc gacacccacg ttccatccat
     2161 aagcctaagt ccgaggaccc cacacagttt cagtttgcac gcccccttga gggacttata
     2221 cagacccctc tcatcgacaa atccgcacgt ttctgacccc tttgatgccc tctttcctct
     2281 tgagttcgtg aaaatgtcac atgacattcc gagtctcgga ttctccggca tccagatcgt
     2341 ataatcgtga ttggtcgaac aatacgtgct gctgacggct acccccgagc attttccgct
     2401 aggaaacacg cgcgagtgga gggaccgatc atacgggtcc aaatccgcga ccgacggcga
     2461 aatgattacg agcgactctt tcgttgtttt tactgtcctc aaccagcggt aatcaggata
     2521 ggggttatgc agtgattctt cgtagcgcgg gtccccagcc atcttccagt tgtatgcagc
     2581 ccgacaggcg tccggcgtgg ggcgaaaatg ctttcttttg aaggtagtag tcacatagcc
     2641 cacgaagttc gtgtatgtct ccgcctccgt caccacgccg gtacacgtga acccattgac
     2701 tttgatggcc aggatgtatc cgaccttgag ttccatgtac gagaaccccg acaagttggt
     2761 gcatccctca tcctccacta cgagattgtt ggggcaggac agatgatgaa tatcgatagg
     2821 ggaccagggt cccagtttat caggaattgt gtagataggg aactttccga aacacagggg
     2881 aaacacgagc aaagggacaa acaaaagggc ttgcgggact ccggaaggac cggggttact
     2941 ttcaacatcg ccagcgagtt tcaacaaagc gtagttagta cattgaccac taccgctagc
     3001 ggagaacaag tctgcctggc tcttgtcagg caccagcagc tccttggatg cgctttcaag
     3061 actgaagatg tcagacctgc tttttgcttt ggacttcccc catgcagcga taccacccac
     3121 agctaccagg cagcacagga gcactgcagt gatcagcatc cacatgcggc cgtgattcct
     3181 ggctggcaga gcacgtccag gagcaggggc agtgggagcc tctgctcctg gacgtgggca
     3241 ccgcggggat cgcgctggtc ccgcagcccc actcgtcccg cccgtcgtcg cagtcggggt
     3301 gtccgtcgca cagccagtct tgcgggtaac actccccgtg ggcaccgggc ggctccgagc
     3361 agcggaactg accggggggg caacgggaca aagaaccgtt accggacccg ttaccggtca
     3421 cgttaccggg cagcagcagc agcagcagcg cgggcagcag ccgcgcatcg attgggccag
     3481 gattctcctc gacgtcaccg catgttagca gacttcctct gccctctcca ctaccaagct
     3541 tcttatcatc atcgtctttg tagtccccct tgtcatcatc atccttgtag tcgcctttgt
     3601 cgtcgtcgtc cttgtaatcc ccaccgttga gtttgtggcc gagcttggag ggaaggtcac
     3661 aataccgagc caccgcgact tcgtgctgct cgacgtaagt ctcgttgttc gcctctttga
     3721 tgcgttcaag tcggtagtcg acatagtaca ctccgggcat cttgagattc ttggcaggct
     3781 ttttgctcct atatgtggtc ttggcgttcg caataagatg cgaaccccct accaatttga
     3841 gcgccatatc attcctccct tccagaccac cgtcagccgg gtacagggtt tcagtgaatg
     3901 cctcccatcc gagtgtcttt ttctgcatga ccggaccatt ggatgtgaaa ttgactcctc
     3961 tgatcttcac gttgtagata aggcacccgt cttggagcga cgtgtcttgt gtcgcagtca
     4021 gcactccacc atcctcatac gtcgtcactc tctcccacgt aaacccttcg ggaaatgact
     4081 gcttaaagaa atcaggaatc ccctgcgtgt gattgataaa cgttttcgat ccataaagga
     4141 acgaggttgc cagaatgtcg aaggcaaagg gcaatgggcc tccttccact actttgatcc
     4201 gcatggtctg tgtcccctcg tacggctttc cctccccttc tgacgtacac ttgaaatgat
     4261 ggttatccac tgttccctcc atatacagtt tcatatgcat gttttccttg atcaactcct
     4321 ctcctttgct gaccatggtg gcggcactag tggatccggc cggccttgat aacttcgtat
     4381 aatgtatgct atacgaagtt atccgaatcg caataacttc gtataaagta tcctatacga
     4441 agttatcgaa gatcatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     4501 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     4561 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     4621 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     4681 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     4741 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     4801 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     4861 ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga
     4921 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     4981 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     5041 tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatcgatc tgcctcgact
     5101 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     5161 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     5221 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     5281 gaagacaata gcaggcatgc tggggacacg tgcggaccga gcggccgcag gaacccctag
     5341 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     5401 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     5461 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     5521 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5581 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     5641 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5701 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     5761 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5821 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     5881 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     5941 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     6001 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     6061 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     6121 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     6181 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     6241 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     6301 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     6361 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     6421 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     6481 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     6541 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     6601 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     6661 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6721 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6781 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6841 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6901 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     6961 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     7021 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     7081 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     7141 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     7201 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     7261 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     7321 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     7381 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     7441 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     7501 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     7561 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     7621 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     7681 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     7741 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     7801 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7861 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     7921 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7981 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     8041 cttttgctgg ccttttgctc acatgtcctg caggcagctg cgcgctcgct cgctcactga
     8101 ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca
     8161 gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgcacgcga aacaattctg
     8221 caggaatcta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     8281 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     8341 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     8401 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     8461 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     8521 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     8581 gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc
     8641 tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg
     8701 ggggggggg
//
