LOCUS       sequence-469218-        7151 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre dependent NaChBac-TdTomato expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7151)
  AUTHORS   Kosse C, Ivanov J, Knight Z, Pellegrino K, Friedman J
  TITLE     A subcortical feeding circuit linking an interoceptive node to jaw 
            movement.
  JOURNAL   Nature. 2024 Dec;636(8041):151-161. doi: 10.1038/s41586-024-08098-1.
            Epub 2024 Oct 23.
  PUBMED    39443799
REFERENCE   2  (bases 1 to 7151)
  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/s41586-024-08098-1"; journalName: "Nature"; date: 
            "2024-12"; volume: "636"; issue: "8041"; pages: "151-161"
COMMENT     Sequence Label: pAAV-Syn-DIO-NaChBac-dTomato
FEATURES             Location/Qualifiers
     source          1..7151
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(660..693)
                     /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)."
     protein_bind    complement(744..777)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(786..2216)
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     regulatory      2213..2222
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2232..2285)
                     /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"
     protein_bind    3136..3169
                     /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)."
     protein_bind    3220..3253
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3278..3866
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3331..3351)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3749..3760)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3898..4374
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4097..4116)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4414..4554
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4414..4543
                     /label=AAV2 ITR
     rep_origin      4629..5084
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4716..4735)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4926..4947
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5101..5120)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5220..5242
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5280..5298)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5366..5470
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5471..5539,5540..6331)
                     /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(5689..5708)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6502..7090
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6991..7010
                     /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 gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gactccggaa
      661 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
      721 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
      781 gcgccttact tgtacagctc gtccatgccg tacaggaaca ggtggtggcg gccctcggag
      841 cgctcgtact gttccacgat ggtgtagtcc tcgttgtggg aggtgatgtc cagcttggtg
      901 tccacgtagt agtagccggg cagttgcacg ggcttcttgg ccatgtagat ggtcttgaac
      961 tccaccaggt agtggccgcc gtccttcagc ttcagggcct ggtggatctc gcccttcagc
     1021 acgccgtcgc gggggtacag gcgctcggtg gaggcctccc agcccatggt cttcttctgc
     1081 attacggggc cgtcgggggg gaagttggtg ccgcgcatct tcaccttgta gatcagcgtg
     1141 ccgtcctgca gggaggagtc ctgggtcacg gtcaccagac cgccgtcctc gaagttcatc
     1201 acgcgctccc acttgaagcc ctcggggaag gacagcttct tgtaatcggg gatgtcggcg
     1261 gggtgcttca cgtacgcctt ggagccgtac atgaactggg gggacaggat gtcccaggcg
     1321 aagggcaggg ggccgccctt ggtcaccttc agcttggcgg tctgggtgcc ctcgtagggg
     1381 cggccctcgc cctcgccctc gatctcgaac tcgtggccgt tcatggagcc ctccatgcgc
     1441 accttgaagc gcatgaactc tttgatgacg gccatgttgt tgtcctcgga ggaggcggtg
     1501 ccggagctgc cgctgccggt gctgccggtg ccatgcccca ggaacaggtg gtggcggccc
     1561 tcggagcgct cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt gatgtccagc
     1621 ttggtgtcca cgtagtagta gccgggcagt tgcacgggct tcttggccat gtagatggtc
     1681 ttgaactcca ccaggtagtg gccgccgtcc ttcagcttca gggcctggtg gatctcgccc
     1741 ttcagcacgc cgtcgcgggg gtacaggcgc tcggtggagg cctcccagcc catggtcttc
     1801 ttctgcatta cggggccgtc gggggggaag ttggtgccgc gcatcttcac cttgtagatc
     1861 agcgtgccgt cctgcaggga ggagtcctgg gtcacggtca ccagaccgcc gtcctcgaag
     1921 ttcatcacgc gctcccactt gaagccctcg gggaaggaca gcttcttgta atcggggatg
     1981 tcggcggggt gcttcacgta cgccttggag ccgtacatga actgggggga caggatgtcc
     2041 caggcgaagg gcagggggcc gcccttggtc accttcagct tggcggtctg ggtgccctcg
     2101 taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcat ggagccctcc
     2161 atgcgcacct tgaagcgcat gaactctttg atgacctcct cgcccttgct caccatggtg
     2221 gcgggatccg ctgggccagg attctcctcg acgtcaccgc atgttagcag acttcctctg
     2281 ccctctccac tgccgatatc cttaagttgt ttgagcaagc tcttcagttc ggctacatcc
     2341 tttcgcaatg ctgaaatctc ctgtttaagt ccgtcggcct ccccatcttc ctcattgtcc
     2401 gtaagctccg ccttttccac gttgttcaca atgactccga tgaaaagatt gaaaatgata
     2461 aaagtaccaa tcaggacaaa cgagacaaag tagagccatg accagggcac ctctgcaaaa
     2521 atgggccgca ttacaccgct ggcccagctc tccaaggtca ccacttggaa cagcgtcagc
     2581 aacgaaagct ggaggttccc aaagtattca ggcgatacgt gctggaagag cattgtcccg
     2641 atcactgcaa agatatagaa gaagatggac atcaagatca agatattgcc caacgccgga
     2701 atcgtcatca ccagcgcatc tacgagtctg cgcagggagg ggacgacgga gatcgccctg
     2761 agtacccgca gcactctaag gattcgcaat actgtcacga actgggctcc cgcgaagata
     2821 tgcccagccg cgacaatcag aaaatcaaac caattccacg agctgcggaa gaaggccgac
     2881 tttgggttcg acgccaaaaa cctcatcgcg atctcgatgg tgaagatcca aagcaggacg
     2941 aggtcaatcc gataaaagag ccacttgtgg tccgcgtaga tcctcggata tgtttcgatt
     3001 cccacgatca gtgcattaaa gaggatcagg gcaatgacgg tgaaagtaaa cgctctatgg
     3061 ttgacaattt tctgcatctt tgacgtgaat gagttctgtt tctggcgtgc ttccattttc
     3121 atggtggcgg ctagcataac ttcgtataaa gtatcctata cgaagttatt tgccttaacc
     3181 cagaaattat cactgttatt ctttagaatg gtgcaaagaa taacttcgta taatgtatgc
     3241 tatacgaagt tatgaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     3301 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     3361 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     3421 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     3481 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     3541 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3601 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3661 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct
     3721 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3781 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3841 gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga gatctacggg
     3901 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
     3961 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
     4021 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
     4081 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
     4141 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
     4201 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
     4261 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
     4321 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
     4381 ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     4441 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     4501 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     4561 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     4621 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4681 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     4741 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4801 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     4861 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4921 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     4981 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     5041 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     5101 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     5161 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     5221 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     5281 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     5341 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5401 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5461 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5521 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5581 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5641 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5701 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5761 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5821 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5881 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5941 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6001 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6061 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6121 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6181 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6241 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6301 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6361 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6421 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6481 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6541 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6601 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6661 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6721 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6781 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6841 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6901 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6961 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7021 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7081 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7141 tgctcacatg t
//
