LOCUS       sequence-454424-        3181 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3181)
  TITLE     Accessory plasmids for optogenetics
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3181)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-FLEX-GFP
FEATURES             Location/Qualifiers
     source          1..3181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     18..37
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     277..298
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      340..349
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(366..399)
                     /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(470..503)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    545..1133
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(598..618)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1016..1027)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1176..1297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1187..1206)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1241..1260
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   1459..1588
                     /label=AAV2 ITR
     primer_bind     2515..2537
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2599..2618
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    2653..2686
                     /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    2749..2782
                     /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(2805..3181,1..337,338..340,341..343))
                     /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(2808..3181,1..337,338..340,341..343))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2850..2871)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct
       61 ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt
      121 agcggctgaa gcactgcacg ccgtaggtca gggtggtcac gagggtgggc cagggcacgg
      181 gcagcttgcc ggtggtgcaa atgaacttca gggtcagctt gccgtaggtg gcatcgccct
      241 cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga
      301 tgggcaccac cccggtgaac agctcctcgc ccttgctcac catggtggcg gtaccgaatt
      361 ctagtataac ttcgtatagg atactttata cgaagttatc attgggattc ttcctatttt
      421 gatccaagca tcaccatcga ccctctagtc cagatctcac catcgaccca taacttcgta
      481 tagcatacat tatacgaagt tatgtccctc gaagaggttc gaattcgata tcaagcttat
      541 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
      601 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
      661 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
      721 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
      781 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
      841 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
      901 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
      961 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     1021 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     1081 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     1141 ccgtcgaccc gggcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
     1201 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     1261 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     1321 tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa
     1381 atgtggtaaa atcgataagg atcttcctag agcatggcta cgtagataag tagcatggcg
     1441 ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     1501 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     1561 cggcctcagt gagcgagcga gcgcgcagct ggggagtggc caactccatc actaggggtt
     1621 ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc tctaggaaga
     1681 gtaccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     1741 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     1801 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     1861 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     1921 gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc
     1981 tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg
     2041 gggggggggg ggggcggggg ccaggggggg gggggggggg gggggggggg gggggggggg
     2101 gggggggggg gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg
     2161 cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg
     2221 aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc
     2281 cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg
     2341 ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt
     2401 tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg
     2461 ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg
     2521 gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct
     2581 ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag
     2641 aattggatcc ccataacttc gtataaagta tcctatacga agttatatca aaataggaag
     2701 accaatgctt caccatcgac ccgaattgcc aagcatcacc atcgacccat aacttcgtat
     2761 aatgtatgct atacgaagtt atactaggga tcccccctcg agttttactt gtacagctcg
     2821 tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc
     2881 ttctcgttgg ggtctttgct cagggcggac tgggtgctca ggtagtggtt gtcgggcagc
     2941 agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg
     3001 ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg
     3061 gccatgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg
     3121 ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg
     3181 a
//
