LOCUS       sequence-466780-        7478 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pAAV transfer plasmid with Ef1a promoter driving Cre- and 
            Flp-dependent expression of G protein for rabies N2c delta G.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7478)
  AUTHORS   Mendelsohn AI, Nikoobakht L, Bikoff JB, Costa RM
  TITLE     Segregated basal ganglia output pathways correspond to genetically 
            divergent neuronal subclasses.
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 18:2024.08.28.610136. doi: 
            10.1101/2024.08.28.610136.
  PUBMED    39257765
REFERENCE   2  (bases 1 to 7478)
  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: "bioRxiv 
            [Preprint]. 2024 Sep 18:2024.08.28.610136. doi: 
            10.1101/2024.08.28.610136."
COMMENT     Sequence Label: pAAV-Fon/ConN2cG
FEATURES             Location/Qualifiers
     source          1..7478
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        881..2059
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1112..2050
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2007..2027
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(2066..2099)
                     /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(2150..2183)
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     regulatory      2844..2853
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2858..2891
                     /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    2942..2975
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     protein_bind    2982..3015
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     protein_bind    complement(4113..4146)
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     misc_feature    4255..4843
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4308..4328)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4726..4737)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4875..5351
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5074..5093)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5391..5531
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5391..5520
                     /label=AAV2 ITR
     rep_origin      5606..6061
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5693..5712)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5903..5924
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6078..6097)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6197..6219
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6257..6275)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6343..6447
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6448..6516,6517..7308)
                     /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(6666..6685)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga ggaatctttg
      841 cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg
      901 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
      961 cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
     1021 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
     1081 ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc
     1141 tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct ggctgcagta
     1201 cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc
     1261 ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg
     1321 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat
     1381 ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc
     1441 gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg
     1501 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg
     1561 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat
     1621 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg
     1681 gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg
     1741 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg
     1801 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag
     1861 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg
     1921 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct
     1981 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct
     2041 tccatttcag gtgtcgtgag gatccataac ttcgtatagg atactttata cgaagttatg
     2101 tagctggagc aatttgaaaa gattgttgta ttacctctag ctgcgcttaa taacttcgta
     2161 tagtatacct tatacgaagt tatgtcgacc ctcctgtact cacctgctat tggtaaaaat
     2221 gtcacaaggt gtccttggtc tcggattctc gggcatccaa atggtgtaat catggttagt
     2281 tgagcagtag gtagaggaca ccgttattcc tgagcacttt ccgccaggga agacccttga
     2341 gtgaagggat ttgtcatatg ggtccaaatc tgtcacactt ggggatatga taatgaggga
     2401 ctctttggtg gttcttacag ttcgaagcca gtggtagtcg gggtatggat tgtgtaggga
     2461 ctcttcatat ctggggtcac cggccatctt ccagttatac gcggctctac atgcgtctgg
     2521 ggtggggcgg aaatgctttc tcttgaatgt ggttgtgaca taaccaacaa agttggtgta
     2581 ggtctctgcc tctgtcacaa cacctgtgca agtgaacccg ttcactttga tggctgagat
     2641 gtatcccact ttgagttcca tgtaggagaa ctcggacagg ttggtacatc cttcatcctc
     2701 cacaaccagg ttatttggac agctgagatg gtgtatgtca atagggctcc agggaccaag
     2761 ttcgtctggt atcgtgtaaa tggggaactt cccgaaacac aacgaaaaac ccagaaggag
     2821 tacaaacaaa agaacctgag gaaccatggt ggctagcata acttcgtata aagtatccta
     2881 tacgaagtta tttgccttaa cccagaaatt atcactgtta ttctttagat aggtgcaaag
     2941 aataacttcg tataaggtat actatacgaa gttatggtac cgaagttcct attcttcaaa
     3001 tagtatagga acttcaattg ttattggtgc aaagaatcac ttccttaagt ctttatactg
     3061 cccaggaagt tcctattctt caaaaggtat aggaacttcg gcgcgcctta tagtctgatc
     3121 tcgcctccac tcttatatga ttcccatgaa ggtatgactt ttccgctttg ggaagtgact
     3181 gacacattcc cccctgtccc tccaaaactg cgttgtttcg attctggtcg attggctctt
     3241 ctgcaccatg tcattaggga aaatatcaac accaggccaa tcatggcccc tgcagtcatc
     3301 aatacatact ttccccagtt cgggagaccc aggtcaaccc ctgagatctg tttgtacaca
     3361 tcggggaggt gaacttcaac aaaatcctca gcctcatcac cttctttgaa aactgtagaa
     3421 gggtctgcca gggggtgcat cagggggata actgaagatt ccaacaactc catatgttgc
     3481 tggaggaggg atgattgcat ctctgggatt aggacatggt cgtcaggccc caatattata
     3541 ccattgaaaa acaccccgtt cacatgagga tggcaccttc ctccaacttt caaacaccct
     3601 tttgagggga tgatctcatt ccaggtccgg actgacttgt agtgagcatc agcctccatc
     3661 aaggttttgt tgaatatggt atatgctttt ccaaaccctg ggacaagttt tctcaggtga
     3721 ctgagacgtc tgaaacttac tgacttggtg gtcatgatgg actctaatgc gtccagacat
     3781 tcctctcttt tcttgactaa ctcctccaca acgagatgct caatctcgtc tgagcgaaag
     3841 tcgtgcaaat tcaccaactg atctggaggg caccatttgg tctcatctga tgtttgcatc
     3901 gcgacccatg ttccatccat aagtctaagt ccaagaactc cacataactt gagcctgcat
     3961 gctcctttta gagacttata caggcctctt tcatccacaa agccgcaagt cttgttcccg
     4021 ttggatgctc tcttccccct gtgggacgac atgacttaac cagttaacca tgttctctga
     4081 acatgagaag ggctggccac tctccatcta gagaagttcc tatactattt gaagaatagg
     4141 aacttcttgc cttaacccag aaattatcac tgttattctt tagaatagtg caaagagaag
     4201 ttcctatacc ttttgaagaa taggaacttc gaattcgata tcaagcttat cgataatcaa
     4261 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4321 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4381 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     4441 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     4501 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     4561 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     4621 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat
     4681 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     4741 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     4801 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg
     4861 ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg
     4921 aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt
     4981 ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc
     5041 aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag
     5101 tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc
     5161 agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt
     5221 tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg
     5281 tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct
     5341 tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct
     5401 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     5461 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     5521 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     5581 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5641 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     5701 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     5761 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     5821 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5881 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     5941 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     6001 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     6061 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     6121 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     6181 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     6241 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     6301 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     6361 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6421 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6481 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6541 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6601 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6661 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6721 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6781 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6841 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6901 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6961 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     7021 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     7081 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     7141 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     7201 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     7261 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     7321 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     7381 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7441 tgagcgtcag accccgtaga aaagatcaaa ggatcttc
//
