LOCUS       sequence-444168-        4486 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pharmacologically Selective Actuator Module, chimeric ligand-gated 
            ion channels that can be used to manipulate electrophysiological 
            activity..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4486)
  TITLE     Tonic dendritic GABA release by substantia nigra dopaminergic 
            neurons
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4486)
  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            "Unpublished"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: EF1a-DIO-PSAML-141F,Y115F- GlyR-2A-FusionRed
FEATURES             Location/Qualifiers
     source          1..4486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    30..254
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     243..260
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     misc_feature    287..875
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             393..404
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     802..822
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    908..941
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1012..1045
                     /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)."
     CDS             2367..2423
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2424..3122
                     /codon_start=1
                     /product="monomeric red fluorescent protein optimized for 
                     use as a fusion tag (Shemiakina et al., 2012)"
                     /label=FusionRed
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMPMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFMYGSRTFIKHPPGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKVRGVNFPANGPVMQKKTLGWEASTETMYPADGGLEGACDMAL
                     KLVGGGHLICNLETTYRSKKPATNLKMPGVYNVDHRLERIKEADDETYVEQHEVAVARY
                     STGGAGDGGK"
     protein_bind    complement(3133..3166)
                     /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(3229..3262)
                     /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)."
     promoter        complement(3282..4472)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3291..4237
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(3314..3334)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 gcggccgctc ggtccgcacg tggtccggac catagagccc accgcatccc cagcatgcct
       61 gctattgtct tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga
      121 atgacaccta ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg
      181 agtggcacct tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca
      241 actagaaggc acagtcgagg ctgatcagcg agtcgacggt atcgatgcgg ggaggcggcc
      301 caaagggaga tccgactcgt ctgagggcga aggcgaagac gcggaagagg ccgcagagcc
      361 ggcagcaggc cgcgggaagg aaggtccgct ggattgaggg ccgaagggac gtagcagaag
      421 gacgtcccgc gcagaatcca ggtggcaaca caggcgagca gccaaggaaa ggacgatgat
      481 ttccccgaca acaccacgga attgtcagtg cccaacagcc gagcccctgt ccagcagcgg
      541 gcaaggcagg cggcgatgag ttccgccgtg gcaataggga gggggaaagc gaaagtcccg
      601 gaaaggagct gacaggtggt ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca
      661 gtgcacacca cgccacgttg cctgacaacg ggccacaact cctcataaag agacagcaac
      721 caggatttat acaaggagga gaaaatgaaa gccatacggg aagcaatagc atgatacaaa
      781 ggcattaaag cagcgtatcc acatagcgta aaaggagcaa catagttaag aataccagtc
      841 aatctttcac aaattttgta atccagaggt tgattatcga taagcttcgc gaacctcttc
      901 gagggacata acttcgtata atgtatgcta tacgaagtta tgggtcgatg gtgagatctg
      961 gactagaggg tcgatggtga tgcttggatc aaaataggaa gaatcccaat gataacttcg
     1021 tataaagtat cctatacgaa gttatactag gtaccgccac catgcgctgt tctccaggcg
     1081 gcgtgtggct cgccctggct gcttcccttc tgcacgttag cctgcagggt gagttccagc
     1141 gcaaactgta taaggagctt gttaagaatt ataaccccct ggagcggccg gtcgcaaatg
     1201 attcccagcc actgacagtg tacttcagcc tctccttgct gcagatcatg gacgtggatg
     1261 aaaagaacca ggtgctgacc actaatattt ggttgcagat gtcctggacc gatcactact
     1321 tgcagtggaa tgtgagcgaa tacccaggtg taaagactgt aagattccct gacggccaaa
     1381 tctggaaacc agatatcctg ctgttcaaca gcgcagacga aaggtttgat gcaacatttc
     1441 acaccaacgt gttggtcaat tcttcaggcc actgccagta cttcccccct ggaatcttca
     1501 agtcctcatg ctatatcgac gtccgctggt ttcccttcga cgtccagcac tgcaaactca
     1561 aattcgggag ctggagctac ggcggatgga gcctggatct gcaaatgcag gaggctgaca
     1621 tctctggtta catcccgaat ggggagtggg accttgtggg aatccccggt aaaagaagcg
     1681 agcgatttta tgaatgctgc aaagagccct acccagatgt caccttcaca gtgaccatgc
     1741 ggagacgcat gggttattat ctgatccaaa tgtatatccc aagcttgctt atagtgattt
     1801 tgtcatggat ctccttctgg attaatatgg acgccgctcc agctagggtc ggactgggca
     1861 tcaccacagt gctgacaatg actactcaga gctcaggcag ccgagccagc ttgcccaagg
     1921 tttcttacgt gaaggccatc gatatctgga tggctgtctg ccttctgttt gtcttcagcg
     1981 cactgctgga atacgccgct gtcaattttg tgtctcgaca gcataaagag ctgttgcggt
     2041 tcagaagaaa acgacgccac cacaaagagg atgaggcagg agaaggacgc ttcaacttta
     2101 gcgcctatgg tatgggacct gcttgcctcc aggctaaaga cggaatttcc gtgaagggag
     2161 ccaacaatag caacacaacc aacccacccc ctgctccatc taagagcccg gaggaaatgc
     2221 gcaaactctt tattcagaga gcgaaaaaga tcgacaaaat ctcccggatc ggattcccca
     2281 tggctttcct gattttcaac atgttttatt ggatcatcta caagattgtg cgaagggagg
     2341 acgtacacaa ccagtctgga tccggagcca cgaacttctc tctgttaaag caagcaggag
     2401 acgtggaaga aaaccccggt cccatggtga gcgagctgat taaggagaac atgcccatga
     2461 agctgtacat ggagggcacc gtgaacaacc accacttcaa gtgcacatcc gagggcgaag
     2521 gcaagcccta cgagggcacc cagaccatga gaatcaaggt cgtcgagggc ggccctctcc
     2581 ccttcgcctt cgacatcctg gctaccagct tcatgtacgg cagcagaacc ttcatcaagc
     2641 accctccggg catccccgac ttctttaagc agtccttccc tgagggcttc acatgggaga
     2701 gagtcaccac atacgaagac gggggcgtgc tgaccgctac ccaggacacc agcctccagg
     2761 acggctgcct catctacaac gtcaaggtta gaggggtgaa cttcccagcc aacggccctg
     2821 tgatgcagaa gaaaacactc ggctgggagg cctccaccga gacgatgtac cccgctgacg
     2881 gcggcctgga aggcgcatgt gacatggccc tgaagctcgt gggcgggggc cacctgatct
     2941 gcaaccttga gaccacatac agatccaaga aacccgctac gaacctcaag atgcccggcg
     3001 tctacaacgt ggaccacaga ctggaaagaa tcaaggaggc cgacgatgag acctacgtcg
     3061 agcagcacga ggtggctgtg gccagatact ctactggtgg cgctggtgat ggaggtaaat
     3121 gagctagcta gtataacttc gtatagcata cattatacga agttatgggt cgatggtgat
     3181 gcttggcaat tcgggtcgat ggtgaagcat tggtcttcct attttgatat aacttcgtat
     3241 aggatacttt atacgaagtt atgggaattc tcctagagtt ttcacgacac ctgaaatgga
     3301 agaaaaaaac tttgaaccac tgtctgaggc ttgagaatga accaagatcc aaactcaaaa
     3361 agggcaaatt ccaaggagaa ttacatcaag tgccaagctg gcctaacttc agtctccacc
     3421 cactcagtgt ggggaaactc catcgcataa aacccctccc cccaacctaa agacgacgta
     3481 ctccaaaagc tcgatcgaga actaatcgag gtgcctggac ggcgcccggt actccgtgga
     3541 gtcacatgaa gcgacggctg aggacggaaa ggcccttttc ctttgtgtgg gtgactcacc
     3601 cgcccgctct cccgagcgcc gcgtcctcca ttttgagctc cctgcagcag ggccgggaag
     3661 cggccatctt tccgctcacg caactggtgc cgaccgggcc agccttgccg cccagggcgg
     3721 ggcgatacac ggcggcgcga ggccaggcac cagagcaggc cggccagctt gagactaccc
     3781 ccgtccgatt ctcggtggcc gcgctcgcag gccccgcctc gccgaacatg tgcgctggga
     3841 cgcacgggcc ccgtcgccgc ccgcggcccc aaaaaccgaa ataccagtgt gcagatcttg
     3901 gcccgcattt acaagactat cttgccagaa aaaaagcgtc gcagcaggtc atcaaaaatt
     3961 ttaaatggct agagacttat cgaaagcagc gagacaggcg cgaaggtgcc accagattcg
     4021 cacgcggcgg ccccagcgcc caggccaggc ctcaactcaa gcacgaggcg aaggggctcc
     4081 ttaagcgcaa ggcctcgaac tctcccaccc acttccaacc cgaagctcgg gatcaagaat
     4141 cacgtactgc agccaggggc gtggaagtaa ttcaaggcac gcaagggcca taacccgtaa
     4201 agaggccagg cccgcgggaa ccacacacgg cacttacctg tgttctggcg gcaaacccgt
     4261 tgcgaaaaag aacgttcacg gcgactactg cacttatata cggttctccc ccaccctcgg
     4321 gaaaaaggcg gagccagtac acgacatcac tttcccagtt taccccgcgc cactttctct
     4381 aggcaccggc cggttcaatt gccgacccct ccccccaact tctcggggac tgtgggcgat
     4441 gtgcgctctg cccactgacg ggcaccggag ccacgcgtgc ggccgc
//
