LOCUS       sequence-468899-        7903 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAG-driven Cre recombinase coupled with H2B-GFP for nuclear 
            fluorescent labelling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7903)
  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 7903)
  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-iCRE-2A-H2BGFP-WPRE
FEATURES             Location/Qualifiers
     source          1..7903
                     /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"
     CDS             1918..1938
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2974..3027
                     /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             3046..3423
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSAK"
     CDS             join(3508..3510,3511..4221)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3550..3571)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3811..3830)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4158..4179
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4307..4895
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4360..4380)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4778..4789)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4930..4947)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4936..5143
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5166..5306
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5166..5295
                     /label=AAV2 ITR
     rep_origin      5381..5836
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5468..5487)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5678..5699
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5853..5872)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5972..5994
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6032..6050)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6118..6222
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6223..6291,6292..7083)
                     /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(6441..6460)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7254..7842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7743..7762
                     /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 ggggccaggg
      661 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg ggggggtgga
      721 aaaaaagggg gcggccgggt aaacgttctt ttatgggtgg gggggggtgg gcgggggccc
      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 catggtgccc
     1921 aagaagaaga ggaaagtctc caacctgctg actgtgcacc aaaacctgcc tgccctccct
     1981 gtggatgcca cctctgatga agtcaggaag aacctgatgg acatgttcag ggacaggcag
     2041 gccttctctg aacacacctg gaagatgctc ctgtctgtgt gcagatcctg ggctgcctgg
     2101 tgcaagctga acaacaggaa atggttccct gctgaacctg aggatgtgag ggactacctc
     2161 ctgtacctgc aagccagagg cctggctgtg aagaccatcc aacagcacct gggccagctc
     2221 aacatgctgc acaggagatc tggcctgcct cgcccttctg actccaatgc tgtgtccctg
     2281 gtgatgagga gaatcagaaa ggagaatgtg gatgctgggg agagagccaa gcaggccctg
     2341 gcctttgaac gcactgactt tgaccaagtc agatccctga tggagaactc tgacagatgc
     2401 caggacatca ggaacctggc cttcctgggc attgcctaca acaccctgct gcgcattgcc
     2461 gaaattgcca gaatcagagt gaaggacatc tcccgcaccg atggtgggag aatgctgatc
     2521 cacattggca ggaccaagac cctggtgtcc acagctggtg tggagaaggc cctgtccctg
     2581 ggggttacca agctggtgga gagatggatc tctgtgtctg gtgtggctga tgaccccaac
     2641 aactacctgt tctgccgggt cagaaagaat ggtgtggctg ccccttctgc cacctcccaa
     2701 ctgtccaccc gggccctgga agggatcttt gaggccaccc accgcctgat ctatggtgcc
     2761 aaggatgact ctgggcagag atacctggcc tggtctggcc actctgccag agtgggtgct
     2821 gccagggaca tggccagggc tggtgtgtcc atccctgaaa tcatgcaggc tggtggctgg
     2881 accaatgtga acattgtgat gaactacatc agaaacctgg actctgagac tggggccatg
     2941 gtgaggctgc tcgaggatgg ggacggcagt ggagagggca gaggaagtct gctaacatgc
     3001 ggtgacgtcg aggagaatcc tggcccagga gatctgaata tcaccatgcc agagccagcg
     3061 aagtctgctc ccgccccgaa aaagggctcc aagaaggcgg tgactaaggc gcagaagaaa
     3121 ggcggcaaga agcgcaagcg cagccgcaag gagagctatt ccatctatgt gtacaaggtt
     3181 ctgaagcagg tccaccctga caccggcatt tcgtccaagg ccatgggcat catgaattcg
     3241 tttgtgaacg acattttcga gcgcatcgca ggtgaggctt cccgcctggc gcattacaac
     3301 aagcgctcga ccatcacctc cagggagatc cagacggccg tgcgcctgct gctgcctggg
     3361 gagttggcca agcacgccgt gtccgagggt actaaggccg tcaccaagta caccagcgct
     3421 aagggatctg ggggtggagg ctccggcggg ggtggatctg gaggtggggg cagcggcgga
     3481 ggtgggctag cgctaccgcg ggccaccatg agcaagggcg aggagctgtt caccggggtg
     3541 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     3601 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     3661 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     3721 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3781 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3841 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3901 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3961 atcatggccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     4021 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     4081 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     4141 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactcac
     4201 ggcatggacg agctgtacaa gtccggactc agatcatcaa gctcatcgga tccgggcggc
     4261 cgtgataata ggtgatagta agtaatagtg aggtaagctt atcgataatc aacctctgga
     4321 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     4381 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     4441 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     4501 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     4561 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     4621 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     4681 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct atgttgccac
     4741 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     4801 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     4861 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag
     4921 atcgatctgc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     4981 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     5041 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     5101 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggacacgtgc ggaccgagcg
     5161 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     5221 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     5281 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     5341 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     5401 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     5461 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     5521 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5581 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     5641 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     5701 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     5761 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     5821 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     5881 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     5941 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     6001 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     6061 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     6121 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6181 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6241 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6301 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6361 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     6421 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     6481 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     6541 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     6601 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     6661 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     6721 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     6781 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     6841 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     6901 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     6961 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     7021 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     7081 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     7141 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     7201 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     7261 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     7321 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7381 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     7441 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     7501 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     7561 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     7621 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     7681 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     7741 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     7801 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7861 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
