LOCUS       sequence-403522-        5276 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Neuron specific expression of a Cre-dependent mRuby3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5276)
  AUTHORS   Cohen-Kashi Malina K, Tsivourakis E, Kushinsky D, Apelblat D, 
            Shtiglitz S, Zohar E, Sokoletsky M, Tasaka GI, Mizrahi A, Lampl I, 
            Spiegel I
  TITLE     NDNF interneurons in layer 1 gain-modulate whole cortical columns 
            according to an animal's behavioral state.
  JOURNAL   Neuron. 2021 Jul 7;109(13):2150-2164.e5. doi: 
            10.1016/j.neuron.2021.05.001. Epub 2021 May 25.
  PUBMED    34038743
REFERENCE   2  (bases 1 to 5276)
  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.neuron.2021.05.001"; journalName: "Neuron"; date: 
            "2021-07-7- 7"; volume: "109"; issue: "13"; pages: "2150-2164.e5"
COMMENT     Sequence Label: pAAV-hSyn-Flex-mRuby3-2A
FEATURES             Location/Qualifiers
     source          1..5276
                     /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        166..613
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(646..679)
                     /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(690..723)
                     /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(751..816)
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     regulatory      1545..1554
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1579..1612
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1623..1656
                     /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    1680..2268
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1733..1753)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2151..2162)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2303..2320)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2309..2516
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2539..2679
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2539..2668
                     /label=AAV2 ITR
     rep_origin      2754..3209
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2841..2860)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3051..3072
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3226..3245)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3345..3367
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3405..3423)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3491..3595
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3596..3664,3665..4456)
                     /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(3814..3833)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4627..5215
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5116..5135
                     /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 agggccctgc gtatgagtgc aagtgggttt
      181 taggaccagg atgaggcggg gtgggggtgc ctacctgacg accgaccccg acccactgga
      241 caagcaccca acccccattc cccaaattgc gcatccccta tcagagaggg ggaggggaaa
      301 caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag tgccttcgcc
      361 cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga cgtcactcgc
      421 cggtcccccg caaactcccc ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc
      481 agccggaccg caccacgcga ggcgcgagat aggggggcac gggcgcgacc atctgcgctg
      541 cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg
      601 cctgagagcg cagacgcgtg gacggtaccc ttcaggtcga cgtcgataac ttcgtatagc
      661 atacattata cgaagttatc cgaatcgcaa taacttcgta taggatactt tatacgaagt
      721 tatttggcgc gccgctagct ttcagaattc tggacctgga ttgctttcta catccccagc
      781 cagtttgagt aaatcaaagt taagagtttg tttgacagga gcgacaattt tggatccctt
      841 ataaagttca tccataccac caccaagatt ggagtacttg gcaactgcga cttcgcgttg
      901 tactacgtac gtttcattgt cactttcctc tatcctttcc aggcggtgat caacggcatg
      961 gacaccgggc atcttgatgt tcccgacggt cttttttgac ctgtaagttg ttacgaaatt
     1021 gcaatgcaga tggccaccac catcaacttt cagtgcaata tcagtgtatc ccctcagacc
     1081 accatctgct ggatacatca tctctgtatt aggctcccaa cccttggtct tcttctgcat
     1141 cacgggacca ttggagggaa agtttacccc tctgactttg acgttgtaaa cgagttcgcc
     1201 atcctcaagg ctggtgtcct gcgtgacggt gacgactcca ccatcttcgt atctcgtaac
     1261 tctttcccaa gtaaaaccct caggaaagga ctgtttaaag aaatcaggaa tgtctgccgg
     1321 gtacttgata aaagtacggc tgccatacat gaacgacgtg gcaagaatgt caaaggcaaa
     1381 tggcaggggt cctccctcga tgactttgat cctcatggtt tgcactccct cgtacggcct
     1441 gccttctcct tcacctgtgc atttgaattg gtggccgttg accgaacctt ccatgaccac
     1501 cttcatacgc atattttcct tgatcagctc ttcgccctta gacaccatgg tggcggcgcg
     1561 ccaacctggc cggccttgat aacttcgtat aatgtatgct atacgaagtt atccgaatcg
     1621 caataacttc gtataaagta tcctatacga agttatcgaa gatcatcaag cttatcgata
     1681 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     1741 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     1801 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     1861 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     1921 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     1981 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     2041 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     2101 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     2161 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     2221 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag
     2281 cgctgctcga gagatcgatc tgcctcgact gtgccttcta gttgccagcc atctgttgtt
     2341 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2401 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2461 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggacacg
     2521 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     2581 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     2641 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     2701 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     2761 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2821 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2881 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2941 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     3001 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3061 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     3121 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3181 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     3241 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3301 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3361 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     3421 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     3481 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3541 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3601 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3661 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3721 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3781 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3841 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3901 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3961 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4021 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4081 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4141 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4201 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4261 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4321 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4381 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4441 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4501 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4561 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4621 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4681 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4741 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4801 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4861 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4921 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4981 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5041 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5101 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5161 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5221 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
