LOCUS       sequence-385176-        8107 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV2 transfer plasmid for Cre-dependent expression of iLMO2, an 
            opto-chemogenetic probe for neuronal inhibition by light or 
            chemical.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8107)
  AUTHORS   Jaiswal PB, Tung JK, Gross RE, English AW
  TITLE     Motoneuron activity is required for enhancements in functional 
            recovery after peripheral nerve injury in exercised female mice.
  JOURNAL   J Neurosci Res. 2017 Aug 3. doi: 10.1002/jnr.24109.
  PUBMED    28771790
REFERENCE   2  (bases 1 to 8107)
  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: "J Neurosci 
            Res. 2017 Aug 3. doi: 10.1002/jnr.24109."
COMMENT     Sequence Label: pAAV/EF1a::DIO-iLMO2
FEATURES             Location/Qualifiers
     source          1..8107
                     /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        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1440..1473)
                     /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(1524..1557)
                     /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(2532..2553)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(join(2688..3200,3201..3203))
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIE"
     primer_bind     2881..2900
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3140..3161
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    4092..4125
                     /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    4176..4209
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4234..4822
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4287..4307)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4705..4716)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4854..5330
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5053..5072)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5370..5510
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5370..5499
                     /label=AAV2 ITR
     rep_origin      5585..6040
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5672..5691)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5882..5903
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6057..6076)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6176..6198
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6236..6254)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6322..6426
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6427..6495,6496..7287)
                     /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(6645..6664)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7458..8046
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7947..7966
                     /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 gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa
     1441 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
     1501 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
     1561 gcgccttaca cctcgttctc gtagcagaac tgctcgttct tcagcactct ctccacgaag
     1621 ctcttgatgt acttgcccat ctcgtcgggg gcgtcctcct ggaggaagtg caggcccttc
     1681 accttcacga actcggtgtt ggggaacttc ttggcgccct ccacgatggc gttgctgaag
     1741 aagccggggt cgccctcgat gaacagcttg ggcaggtcgt cgctggctct caggtaggcg
     1801 ttgtagtttc tcacgatctg caccacgtcg ggcttgccgc ccttcaccag ggggatctct
     1861 ctgggccagc tcagggtggg tcttctcacc tcgcccttct ccttgaaggg ctccaggtag
     1921 gcggcgaact cctcgggctc cagctttctc atgatcttgc tgggcagcac ggtctccacg
     1981 aagaagttgt tctccagcac catcttctcg ccctcctcgc tcttgatcag ggcgatgtcc
     2041 tcctcgatgt ctggccactc gtcccagctc tcgatcacgt ccaccacgct ctccatgtgc
     2101 acgatggcct tgatcctgtc ctggtgctcg taggcgtagt ggaaggccag ggcggcgccc
     2161 cagtcgtggc ccacgaagat gatcttcttg ggcaggttca gaagctcgaa ccaggcggtc
     2221 aggtacttgt agtggtccag cagcctgtag ctgccgttgc cgctcttgcc gctcttgccc
     2281 atgccgatca gatcggggat gatgcacctg gccacgggct cgatgtgggg caccacgtgc
     2341 ctccacaggt agctgctagt ggcgttgccg tgcaggaaga tcacggcgtt ctcggcgtgc
     2401 ttctcgctgt cgtagtagtt gatgaagctg tccagcacgt tcatctgctt gcacctggcc
     2461 caccactggg ggccggtgat catcctcttc ctctgctcgg ggtcgtacac cttggtaccc
     2521 ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg
     2581 ctcagcttgg actggtagct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg
     2641 atgggggtgt tctgctggta gtggtcggcg agctgcacgc cgccgtcctc gatgttgtgg
     2701 cggatcttga agttggcctt gatgccgttc ttctgcttgt cggcggtgat atagacgttg
     2761 tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg
     2821 atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg
     2881 gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc
     2941 atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc
     3001 aggccgtagc ccagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg
     3061 cagatcagct tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg
     3121 ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg
     3181 aacagctcct cgcccttgct cactgcggcc gcatcatcag ccggggtccc agaagcagat
     3241 ggaacatcca aaatgctgcc ggacacgaca gattcgtttg acgtcagata gttcaggagc
     3301 aagaatgcga aaatgtactt tgccacgata tccaggaaag aatatcccca gctcgtcacc
     3361 ccaacgggca acaccgcgat accctcgact ccaagagccc acacaattgg ataccccagc
     3421 cacatcacta ctgtcaacag cttcagggta ttgaacatgt cagcggttcc cgcggctttg
     3481 gcgtcctgtg cccactccac cagcaggata tacaagacca ccagaaagca tgcacaactg
     3541 atagcgtacc agaaccatct catgaggtgg gaggaggtag tcagggccgc ggcaaggcca
     3601 gtcacgcaca tagcgatatc gaaagtgata gctgtgaaga gctttgtagc attgcttccg
     3661 gccaggagac ccagagcgag gagaatcatg ggcgtggaaa gtgcccaggt gagataccgt
     3721 ccccacatgg tgactacacc atctacctct tctcctccca gcatcactga gctgccttct
     3781 gcaaaatggc ccgctggcat ttcaagaacg gaaattgtca ggccgctcgc caatccagtg
     3841 taggaggcaa tgctgacgac aggcacaagg atggttgaca cagcaataag ttttgcccgt
     3901 ggatcatcga gtcctcgggt cataaaaacg aacagcagta tactcagtcc tgcaagtgcg
     3961 atgttgatat agagactgct tgcaagcaaa gggtcgttca gcacgaactc gaacaactcc
     4021 ctttgggtaa cctcggcttg aagggccacg gcactctcgg tcacgggagg cagggtctct
     4081 gtcatgctag cataacttcg tataaagtat cctatacgaa gttatttgcc ttaacccaga
     4141 aattatcact gttattcttt agaatggtgc aaagaataac ttcgtataat gtatgctata
     4201 cgaagttatg aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt
     4261 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     4321 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     4381 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     4441 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     4501 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     4561 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     4621 tcggggaaat catcgtcctt tccttggctg ctcgcctatg ttgccacctg gattctgcgc
     4681 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     4741 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     4801 tccctttggg ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc tacgggtggc
     4861 atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca
     4921 ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata
     4981 atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta
     5041 gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg
     5101 caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat
     5161 tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca
     5221 ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc
     5281 ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt
     5341 aggtaaccac gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc
     5401 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     5461 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat
     5521 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     5581 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     5641 accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc
     5701 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     5761 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     5821 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5881 agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat
     5941 ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa
     6001 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     6061 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     6121 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     6181 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     6241 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     6301 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     6361 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     6421 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     6481 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     6541 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     6601 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     6661 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     6721 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     6781 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     6841 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     6901 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6961 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     7021 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     7081 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     7141 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     7201 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     7261 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     7321 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     7381 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     7441 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     7501 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     7561 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     7621 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     7681 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     7741 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     7801 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     7861 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     7921 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     7981 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     8041 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     8101 cacatgt
//
