LOCUS       sequence-468900-        7578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAG-driven, Cre-dependent FlpO recombinase for H2B-V5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7578)
  AUTHORS   Falasconi A, Kanodia H, Arber S
  TITLE     Dynamic basal ganglia output signals license and suppress forelimb 
            movements.
  JOURNAL   Nature. 2025 May 28. doi: 10.1038/s41586-025-09066-z.
  PUBMED    40437098
REFERENCE   2  (bases 1 to 7578)
  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 May 28. doi: 10.1038/s41586-025-09066-z."
COMMENT     Sequence Label: pAAV-pCAG-Floxed-FlpO-2A-H2BV5c.o.,rev.-WPRE
FEATURES             Location/Qualifiers
     source          1..7578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        223..526
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          808..1824
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1748..1770
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1832..1851
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1928..1961)
                     /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(1972..2005)
                     /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(2051..2092)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(2486..2539)
                     /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(3818..3838)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      3841..3850
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3881..3914
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3925..3958
                     /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    3982..4570
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4035..4055)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4453..4464)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4605..4622)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4611..4818
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4841..4981
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4841..4970
                     /label=AAV2 ITR
     rep_origin      5056..5511
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5143..5162)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5353..5374
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5528..5547)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5647..5669
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5707..5725)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5793..5897
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5898..5966,5967..6758)
                     /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(6116..6135)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6929..7517
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7418..7437
                     /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 gcgaaacaat tctgcaggaa tctagttatt aatagtaatc
      181 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      241 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      301 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      361 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      421 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      481 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
      541 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
      601 tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggg gggcgcgggg
      661 gggggggggg gggggggggg gggggggggc ggggcgaggc ggagaggtgc ggcggcagcc
      721 aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc
      781 tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc
      841 cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag
      901 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg
      961 cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc
     1021 ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc
     1081 tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc
     1141 agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag
     1201 gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg
     1261 tcggtcgggc tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct
     1321 tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg
     1381 cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag
     1441 gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc
     1501 cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag
     1561 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc
     1621 gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt
     1681 ctccctctcc agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc
     1741 agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt
     1801 catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat
     1861 cattttggca aagaattgat ttgataccgc gggccccgcg tggacggtac ccttcaggtc
     1921 gacgtcgata acttcgtata gcatacatta tacgaagtta tccgaatcgc aataacttcg
     1981 tataggatac tttatacgaa gttatttggc gcgccgctag ctcactatta cttactatca
     2041 cctattatca cgtgctgtcc aggcccagca aaggattagg gatgggcttg ccggcgctag
     2101 tgtacttagt cactgcctta gttccctcag atacagcgtg cttagccagt tcgccaggga
     2161 ggagaagtct cacagctgtc tgaatttcgc gtgatgtgat cgtagacctc ttgttgtagt
     2221 gggccaatcg gcttgcctcc ccagctattc tctcgaatat gtcgttcacg aagctattca
     2281 ttatacccat cgcttttgat gaaatcccag tgtcagggtg tacctgcttc agcactttgt
     2341 aaacgtatat agagtaggac tctttccttg accgtttccg ttttttaccg cctttcttct
     2401 gggccttggt cactgctttt ttggaaccct tcttcggcgc aggagcactc ttagcaggtt
     2461 caggcattgt gatattcagg tcgccggggc caggattctc ctccacgtcg ccgcaagtca
     2521 gaaggctccc ccgtccctcg ccggatccga tccgtctatt aatataagag gagaggtagt
     2581 cgaggacttc ctgactgatg attccgttcc aagctgggta cctaatgctc ccctcagcgg
     2641 atcccttgag ctgttcaata tgctgccatt cctcaatggg atttgtttca tccttgaggg
     2701 cgatcatctc tttggagata ggatcgtaag cgtaatatct gctaaccaga gcgaaatagt
     2761 gatcaggaat agcggtgatc tgatgtgtgt acgtggttct agcaacagca ctggccctct
     2821 tgtcggacca gttgccaacg acattagtga gctctgtcaa acctttcata ctgagaaagc
     2881 tagtcatcaa gtgccggcca atgtgggatt ttggtccatt tttgatagca aaaattggat
     2941 aaggagcgtt cttcttaaga gctttgttat agctgcggac cagattatct ttcagcagct
     3001 ggtactcttg cttattagat gatgaattgc cggttcggtt caccctcttc agcactggct
     3061 cgctgttccg cagaaattca tccaagtaca cgagggggtc aattcgaccg cgtgcgctaa
     3121 agaagtagat gtggcggctc actgaagtct tggtctcggt gaccaggcat tggattataa
     3181 cgccgaggta cttgttctgc accagcttga agctcttagg gtctacgttc ttaatgtcgg
     3241 aaaaccgacc gcagttaatg aaggttgcca gaaacaggaa ttgatacaga gtttttgtct
     3301 ttgtgaatcg actcgtgtat tcgaaagaat tcaggatttt ttctgtaatc tcccaaatag
     3361 actcgccctc ggacagcaat gccttgagca ttttcttgga gtggctattc cccttatctg
     3421 cttcctcgct agactcgaac tgcagctgca aagaacttac tatatcagtt atatcgcttt
     3481 gatgtttctg accattgtag ggtatgatag tgaactccca ggcgggaatg agcttcttga
     3541 gtgaagcctc caggattgtg gctttctggg tcttatattt aaactgcagg cttttattaa
     3601 cgatgtcgaa ggacagagag tttgaaatga ttgtattgta ggacataaaa gtagcccgct
     3661 ttatagcagt tccgttatga gtaatcatcc agcaaaggta tgtcagctca gcggcgcagg
     3721 aggcaatttt ctcgccggat ggccgttcaa acctctcgac aaactgtctc acgagaacct
     3781 tcgggggggt tttgcacagg atgtcaaact gtgacatgac tttgcgcttc ttctttgggg
     3841 ccatggtggc gctagcggcg cgccaacctg gccggccttg ataacttcgt ataatgtatg
     3901 ctatacgaag ttatccgaat cgcaataact tcgtataaag tatcctatac gaagttatcg
     3961 aagatcatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     4021 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     4081 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     4141 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     4201 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     4261 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     4321 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     4381 tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc
     4441 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     4501 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     4561 gcctccccgc atcgataccg agcgctgctc gagagatcga tctgcctcga ctgtgccttc
     4621 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     4681 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     4741 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     4801 tagcaggcat gctggggaca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     4861 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     4921 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     4981 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     5041 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     5101 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     5161 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     5221 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     5281 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5341 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     5401 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     5461 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     5521 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     5581 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     5641 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     5701 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     5761 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5821 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5881 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5941 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     6001 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     6061 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     6121 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     6181 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     6241 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6301 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6361 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6421 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6481 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6541 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6601 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6661 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6721 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6781 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6841 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6901 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6961 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     7021 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     7081 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     7141 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     7201 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     7261 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7321 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7381 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7441 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7501 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7561 ggccttttgc tcacatgt
//
