LOCUS       sequence-444384-        6732 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses VGF and mCherry in a Cre-dependent manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6732)
  AUTHORS   Selten M, Bernard C, Mukherjee D, Hamid F, Hanusz-Godoy A, Oozeer F,
            Zimmer C, Marin O
  TITLE     Regulation of PV interneuron plasticity by neuropeptide-encoding 
            genes.
  JOURNAL   Nature. 2025 Apr 30. doi: 10.1038/s41586-025-08933-z.
  PUBMED    40307547
REFERENCE   2  (bases 1 to 6732)
  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"; journalName: "Nature. 
            2025 Apr 30. doi: 10.1038/s41586-025-08933-z."
COMMENT     Sequence Label: pAAV-hSyn-DIO-Vgf-T2A-mCherry-CW3SL
FEATURES             Location/Qualifiers
     source          1..6732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     38..57
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   199..328
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        373..820
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(858..891)
                     /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(942..975)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2835..2888)
                     /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(2898..3605)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3173..3192)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3442..3460
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3602..3611
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3626..3659
                     /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    3710..3743
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3813..3833)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4193..4333
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4193..4322
                     /label=AAV2 ITR
     rep_origin      4408..4863
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4495..4514)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4705..4726
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4880..4899)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4999..5021
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5059..5077)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5145..5249
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5250..5318,5319..6110)
                     /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(5468..5487)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6281..6732,1..137)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
       61 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
      121 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
      181 ggccttttgc tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc
      241 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      301 gtggccaact ccatcactag gggttcctgc ggccgcacgc gtgtgtctag actgcagagg
      361 gccctgcgta tgagtgcaag tgggttttag gaccaggatg aggcggggtg ggggtgccta
      421 cctgacgacc gaccccgacc cactggacaa gcacccaacc cccattcccc aaattgcgca
      481 tcccctatca gagaggggga ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc
      541 ttcagcaccg cggacagtgc cttcgccccc gcctggcggc gcgcgccacc gccgcctcag
      601 cactgaaggc gcgctgacgt cactcgccgg tcccccgcaa actccccttc ccggccacct
      661 tggtcgcgtc cgcgccgccg ccggcccagc cggaccgcac cacgcgaggc gcgagatagg
      721 ggggcacggg cgcgaccatc tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc
      781 ggtgggcagc ggaggagtcg tgtcgtgcct gagagcgcag tcgagaaggt accggatcct
      841 ctagagtcga ctccggaata acttcgtata ggatacttta tacgaagtta tgcagaatgg
      901 tagctggatt gtagctgcta ttagcaatat gaaacctctt aataacttcg tatagcatac
      961 attatacgaa gttatggcgc gcctcacggg cggtgcagca gcacgtgctc aatgtaattc
     1021 tccagctcct cctgctcctg cagccggcgc tcctccgcgt ccgcttcctc ctgcgcgcgc
     1081 ctggcctggg cctccagatc cggatggtgg cgcgcaggtg gcaacgcgtg atggaagtga
     1141 cggcggcggg aggatgcggg cggctgcagt gtccgcggcc gaatgtagtt tgggaagggg
     1201 tgatagggcc ccgggggaaa cacctcatcc tcctcccgat cccagggtgg gagcacttcg
     1261 ttccagtccg gcaactcatc ccgggctggg gcgggaggtg ggggctgcgg agagcggaca
     1321 tgggtcgggg ctggggcagc cctgggaggc ggcaccggct cgggaggggc gttcttcttc
     1381 cgcttccgtt tctcctccac ctcttcgatg atgctgacca catcgtctgc tggcaggtgg
     1441 agtttggtgg acagttcaat gagactatcg atcgtctgcg gatccatctc ctcgtcgtcg
     1501 tcatcctcct ccccgccctc ctctgccccc tcagcatcct tccgccgatg gcctggcgcc
     1561 tcctcctggg agcgtttgtc ctcggctccg gcttccccgt cctcctcctc ggcgaacagg
     1621 agtgcgttct gccgcgccct ctccgcctcc tctgcctccg cctccgcctc cgccgcctcc
     1681 tcatcctctt cccccacatc gtcctcccca ccacctcgcc tctcctgctc tgcctcctcc
     1741 tccctctcgt tctcccgctc ttgctgcgtc tcctgcagct cgcgaccccc gagatcgcgc
     1801 tgccgggctc cgccctgcaa cagatactgg agcagcaggt cggaggcgag atcggccagc
     1861 cgctcttctt gcgccgcggc ctgccgggtg gcctccgcct gtctcctccc tgcctctacc
     1921 tgagctaatc cttgctgaag caggcgctct cctgcctcgg agccgcccag gaatgagccc
     1981 tcgagccggc gcaccttagg gaaggggcca cctagacttt gatacgcctt ggataagggt
     2041 gtcaaagtct cacctaaatg tgttttaggg gaggacactc cttccccgaa ctgatgagtt
     2101 tcgggaaggg gagcgctttc ggacattcga gcctggaatt gggaagggac cgggggcgga
     2161 agaggagcgc gctccggaac gcgcgcctgg aactctcccc aggaagcgcg ccacacgcgc
     2221 tctggcccgg ggctctccag attgactcgg gtcagcgtgt gcgtgcgggt ttcagtctct
     2281 gctgccgccg tctcttgttg gcgcttagca ttgctcggac tgaaatctcg aagttcttgg
     2341 agcaaggatg ctagcgcctc cagctcttct gagcggtcgt cctcctcggg atcgcggtcc
     2401 tgagtctgag ggcgaggggg ggccacagcg ggcgcttgga tttctggtgc cgggaggcta
     2461 tgggtctgac tgcgcacgga ctcggtcagc agagcttctg ctgcttcttc gggtgttccc
     2521 tgctgggaac ctcccgggac cgacgggggc gaggccggac ggtccagtgc ctgcaacagt
     2581 accgaggcca gcgcccgggg atccacgccc tggaagagct ctccctggtc ctgaggctcg
     2641 ggattccgag cagctcggac ctctggtacg ccgtcatcct ttggccggga cactgcgtcc
     2701 tcagctacct gcccattatg ctcagagctg aggggaggag gaaaaacatc ggggcgcccg
     2761 gggggcgctg ctcccaaccc ctggatcagt agaaggaagc agaagaggac ggatgccggc
     2821 aacgtgaagg ttttggggcc ggggttctcc tccacgtcgc cgcaggtcag caggctgccc
     2881 ctgccctcac cactaccctt gtacagctcg tccatgccgc cggtggagtg gcggccctcg
     2941 gcgcgttcgt actgttccac gatggtgtag tcctcgttgt gggaggtgat gtccaacttg
     3001 atgttgacgt tgtaggcgcc gggcagctgc acgggcttct tggccttgta ggtggtcttg
     3061 acctcagcgt cgtagtggcc gccgtccttc agcttcagcc tctgcttgat ctcgcccttc
     3121 agggcgccgt cctcggggta catccgctcg gaggaggcct cccagcccat ggtcttcttc
     3181 tgcattacgg ggccgtcgga ggggaagttg gtgccgcgca gcttcacctt gtagatgaac
     3241 tcgccgtcct gcagggagga gtcctgggtc acggtcacca cgccgccgtc ctcgaagttc
     3301 atcacgcgct cccacttgaa gccctcgggg aaggacagct tcaagtagtc ggggatgtcg
     3361 gcggggtgct tcacgtaggc cttggagccg tacatgaact gaggggacag gatgtcccag
     3421 gcgaagggca gggggccacc cttggtcacc ttcagcttgg cggtctgggt gccctcgtag
     3481 gggcggccct cgccctcgcc ctcgatctcg aactcgtggc cgttcacgga gccctccatg
     3541 tgcaccttga agcgcatgaa ctccttgatg atggccatgt tatcctcctc gcccttgctc
     3601 accatggtgg cttaattaag ctagcataac ttcgtataaa gtatcctata cgaagttatt
     3661 tgccttaacc cagaaattat cactgttatt ctttagaatg gtgcaaagaa taacttcgta
     3721 taatgtatgc tatacgaagt tatgaattcc gctcgagata atcaacctct ggattacaaa
     3781 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3841 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3901 ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg ccttgcccgc
     3961 tgctggacag gggctcggct gttgggcact gacaattccg tggtgtttat ttgtgaaatt
     4021 tgtgatgcta ttgctttatt tgtaaccatc tagctttatt tgtgaaattt gtgatgctat
     4081 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     4141 ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcggcc gcaggaaccc
     4201 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     4261 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     4321 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4381 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     4441 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     4501 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4561 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4621 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4681 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     4741 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     4801 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     4861 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     4921 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     4981 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     5041 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     5101 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5161 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5221 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5281 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5341 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5401 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5461 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5521 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5581 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5641 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5701 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5761 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5821 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5881 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5941 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6001 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6061 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     6121 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6181 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6241 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6301 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6361 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6421 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6481 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6541 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6601 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6661 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6721 cggtaagcgg ca
//
