LOCUS       sequence-271167-        5465 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Can be used to drive axon-enriched expression of EGFP in the 
            presence of Cre recombinase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5465)
  TITLE     AAV plasmids for the expression of EGFP
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5465)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-hSyn-Flex-Axon-EGFP
FEATURES             Location/Qualifiers
     source          1..5465
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   74..214
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   74..203
                     /label=AAV2 ITR
     rep_origin      289..744
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(376..395)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     586..607
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(761..780)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     880..902
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(940..958)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1026..1130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1131..1199,1200..1991)
                     /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(1349..1368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2162..2750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2651..2670
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2812..2941
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        2994..3441
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(3483..3516)
                     /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(3567..3600)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(3615..4328,4329..4331,4332..4334))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(3618..4328,4329..4331,4332..4334))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3660..3681)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4009..4028
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4268..4289
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4331..4340
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(join(4347..4361,4362..4370))
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             complement(4374..4406)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             complement(4410..4427)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    4540..4573
                     /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    4624..4657
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4688..5276
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4741..4761)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5159..5170)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5310..5421
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 ggcaggacag caagggggag gattgggaag acaatagcag gcatgagatc tcacgtgcgg
       61 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
      121 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
      181 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
      241 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
      301 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
      361 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
      421 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
      481 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
      541 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
      601 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
      661 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
      721 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
      781 catagttacg ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
      841 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
      901 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
      961 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     1021 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1081 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     1141 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     1201 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     1261 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     1321 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     1381 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     1441 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     1501 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     1561 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     1621 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     1681 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     1741 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     1801 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     1861 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     1921 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     1981 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     2041 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     2101 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2161 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2221 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2281 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     2341 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2401 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2461 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2521 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2581 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2641 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2701 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2761 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc
     2821 agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg
     2881 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
     2941 tgcggccgca cgcgtttaat taagtgtcta gactgcagag ggccctgcgt atgagtgcaa
     3001 gtgggtttta ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac
     3061 ccactggaca agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg
     3121 aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg
     3181 ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg
     3241 tcactcgccg gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc
     3301 gccggcccag ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat
     3361 ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc
     3421 gtgtcgtgcc tgagagcgca gtcgagaaac cggctagagg atcctctaga gtcgactccg
     3481 gaataacttc gtataggata ctttatacga agttatgcag aatggtagct ggattgtagc
     3541 tgctattagc aatatgaaac ctcttaataa cttcgtatag catacattat acgaagttat
     3601 ggcgcgccct attattactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc
     3661 acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac
     3721 tgggtgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc
     3781 tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag
     3841 ttcaccttga tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag
     3901 ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc
     3961 tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg
     4021 ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg
     4081 aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac gccgtaggtc
     4141 agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc
     4201 agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg
     4261 ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg
     4321 cccttgctca ccatggtggc gagatcctta tcgtcatcgt cgtacagatc ccgacccatt
     4381 tgctgtccac cagtcatgct agccatacca tgatgatgat gatgatgaga accaatcttt
     4441 tggtcctcat cattcttttc aacctgtttg gttcttctca tacagcacag catgccaccg
     4501 gtggcgggtg gatccttgct agcagcttga attgctagca taacttcgta taaagtatcc
     4561 tatacgaagt tatttgcctt aacccagaaa ttatcactgt tattctttag aatggtgcaa
     4621 agaataactt cgtataatgt atgctatacg aagttatgat atcgaattcg atatcaagct
     4681 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4741 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4801 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4861 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4921 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4981 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     5041 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     5101 gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     5161 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     5221 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     5281 ataccgagcg ctgctcgaga gatcttcgac tgtgccttct agttgccagc catctgttgt
     5341 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     5401 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     5461 ggtgg
//
