LOCUS       sequence-442750-        5222 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Cre-dependently expresses mCherry-CAAX in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5222)
  TITLE     Nagai Lab plasmids for expression in astrocytes and neurons
REFERENCE   2  (bases 1 to 5222)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: hSyn1-DIO-mCherry-CAAX
FEATURES             Location/Qualifiers
     source          1..5222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     protein_bind    137..157
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        179..626
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    626..650
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(651..684)
                     /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(717..750)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(781..1488)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1056..1075)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1325..1343
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1485..1494
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1501..1534
                     /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    1567..1600
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(1601..1625)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1635..2223
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1688..1708)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2106..2117)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2253..2270)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2259..2466
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2474..2614
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2474..2603
                     /label=AAV2 ITR
     rep_origin      2689..3144
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2776..2795)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2986..3007
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3161..3180)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3280..3302
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3340..3358)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3426..3530
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3531..3599,3600..4391)
                     /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(3749..3768)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4562..5150
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5051..5070
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(5212..5222,1..130)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tctagacaac tttgtataga aaagttgctg cagagggccc tgcgtatgag
      181 tgcaagtggg ttttaggacc aggatgaggc ggggtggggg tgcctacctg acgaccgacc
      241 ccgacccact ggacaagcac ccaaccccca ttccccaaat tgcgcatccc ctatcagaga
      301 gggggagggg aaacaggatg cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga
      361 cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc
      421 tgacgtcact cgccggtccc ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg
      481 ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg
      541 accatctgcg ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag
      601 gagtcgtgtc gtgcctgaga gcgcagcaag tttgtacaaa aaagcaggct ataacttcgt
      661 ataggatact ttatacgaag ttatcctcga gtaccggtca cgcgtacgca agcttgataa
      721 cttcgtatag catacattat acgaagttat cctagggcca ccttacagga tgatgcagca
      781 cttgtacagc tcgtccatgc cgccggtgga gtggcggccc tcggcgcgtt cgtactgttc
      841 cacgatggtg tagtcctcgt tgtgggaggt gatgtccaac ttgatgttga cgttgtaggc
      901 gccgggcagc tgcacgggct tcttggcctt gtaggtggtc ttgacctcag cgtcgtagtg
      961 gccgccgtcc ttcagcttca gcctctgctt gatctcgccc ttcagggcgc cgtcctcggg
     1021 gtacatccgc tcggaggagg cctcccagcc catggtcttc ttctgcatta cggggccgtc
     1081 ggaggggaag ttggtgccgc gcagcttcac cttgtagatg aactcgccgt cctgcaggga
     1141 ggagtcctgg gtcacggtca ccacgccgcc gtcctcgaag ttcatcacgc gctcccactt
     1201 gaagccctcg gggaaggaca gcttcaagta gtcggggatg tcggcggggt gcttcacgta
     1261 ggccttggag ccgtacatga actgagggga caggatgtcc caggcgaagg gcagggggcc
     1321 acccttggtc accttcagct tggcggtctg ggtgccctcg taggggcggc cctcgccctc
     1381 gccctcgatc tcgaactcgt ggccgttcac ggagccctcc atgtgcacct tgaagcgcat
     1441 gaactccttg atgatggcca tgttatcctc ctcgcccttg ctcaccatgg tggctctaga
     1501 ataacttcgt ataaagtatc ctatacgaag ttataggtac ctgagctcaa catcgatagc
     1561 actagtataa cttcgtataa tgtatgctat acgaagttat acccagcttt cttgtacaaa
     1621 gtgggaattc cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     1681 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     1741 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     1801 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     1861 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     1921 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     1981 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     2041 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     2101 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     2161 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     2221 cgcatcggga attcctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     2281 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2341 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     2401 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagagaata gcaggcatgc
     2461 tggggagggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     2521 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     2581 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     2641 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     2701 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2761 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2821 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2881 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     2941 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3001 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     3061 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3121 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     3181 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3241 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3301 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     3361 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     3421 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3481 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3541 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3601 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3661 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3721 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     3781 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3841 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3901 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3961 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4021 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4081 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4141 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4201 cggctggctg gtttattgct gataaatctg gagccggtga gcgtggaagc cgcggtatca
     4261 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4321 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4381 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4441 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4501 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4561 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4621 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4681 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4741 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4801 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4861 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4921 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4981 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5041 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5101 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5161 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc
     5221 ag
//
