LOCUS       sequence-140259-        7552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent expression of PSAM-5HT3 HC (L141F,Y115F) neuronal 
            activator, with IRES-EGFP marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7552)
  AUTHORS   Magnus CJ, Lee PH, Atasoy D, Su HH, Looger LL, Sternson SM
  TITLE     Chemical and genetic engineering of selective ion channel-ligand 
            interactions.
  JOURNAL   Science. 2011 Sep 2;333(6047):1292-6.
  PUBMED    21885782
REFERENCE   2  (bases 1 to 7552)
  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: "Science."; 
            date: "2011-09-2"; volume: "333(6047)"; pages: "1292-6"
COMMENT     Sequence Label: rAAV-syn::FLEX-rev::PSAML141F,Y115F:5HT3HC-IRE...
FEATURES             Location/Qualifiers
     source          1..7552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   51..180
                     /label=AAV2 ITR
     intron          773..905
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     888..907
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    918..951
                     /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    1014..1047
                     /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(1058..1771,1772..1774))
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1103..1124)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1452..1471
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1711..1732
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    1798..2349
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     2188..2205
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     protein_bind    complement(3824..3857)
                     /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(3928..3961)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4033..4621
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4086..4106)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4504..4515)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4699..4820
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4710..4729)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4764..4783
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5326..5914)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5406..5425)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6085..6876,6877..6945))
                     /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     6708..6727
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6946..7050)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7076..7531)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7213..7234)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7425..7444
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctgcgcgctc
       61 gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg
      121 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      181 tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct aggaagatcc
      241 gagctccagt gtgctggaat tcgccctttt cagtatttaa attgacgacc gaccccgacc
      301 cactggacaa gcacccaacc cccattcccc aaattgcgca tcccctatca gagaggggga
      361 ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc ttcagcaccg cggacagtgc
      421 cttcgccccc gcctggcggc gcgcgccacc gccgcctcag cactgaaggc gcgctgacgt
      481 cactcgccgg tcccccgcaa actccccttc ccggccacct tggtcgcgtc cgcgccgccg
      541 ccggcccagc cggaccgcac cacgcgaggc gcgagatagg ggggcacggg cgcgaccatc
      601 tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc ggaggagtcg
      661 tgtcgtgcct gagagcgcag ctgtgctcct gggcaccgcg cagtccgccc ccgcggctcc
      721 tggccagacc acccctagga ccccctgccc caagtcgcag ccgttggatc aggtaagtat
      781 caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag
      841 actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc
      901 cacaggtgat gatccccata acttcgtata aagtatccta tacgaagtta tatcaaaata
      961 ggaagaccaa tgcttcacca tcgacccgaa ttgccaagca tcaccatcga cacataactt
     1021 cgtataatgt atgctatacg aagttatact agctagatta cttgtacagc tcgtccatgc
     1081 cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt
     1141 tggggtcttt gctcagcttg gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg
     1201 ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg ctgccgtcct
     1261 cgatgttgtg gcggatcttg aagttcacct tgatgccgtt cttctgcttg tcggccatga
     1321 tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct
     1381 ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca
     1441 cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt
     1501 agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc
     1561 tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc acgggcagct
     1621 tgccggtggt gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcgccct
     1681 cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca
     1741 ccaccccggt gaacagctcc tcgcccttgc tcaccggatc ccccatggtt gtggccatat
     1801 tatcatcgtg tttttcaaag gaaaaccacg tccccgtggt tcggggggcc tagacgtttt
     1861 tttaacctcg actaaacaca tgtaaagcat gtgcaccgag gccccagatc agatcccata
     1921 caatggggta ccttctgggc atccttcagc cccttgttga atacgcttag gagagccatt
     1981 tgactctttc cacaactatc caactcacaa cgtggcactg gggttgtgcc gcctttgcag
     2041 gtgtatctta tacacgtggc ttttggccgc agaggcacct gtcgccaggt ggggggttcc
     2101 gctgcctgca aagggtcgct acagacgttg tttgtcttca agaagcttcc agaggaactg
     2161 cttccttcac gacattcaac agaccttgca ttcctttggc gagaggggaa agacccctag
     2221 gaatgctcgt caagaagaca gggccaggtt tccgggccct cacattgcca aaagacggca
     2281 atatggtgga aaataacata tagacaaacg cacaccggcc ttattccaag cggcttcggc
     2341 cagtaacgtt aggggggggg gggcgggggg aattgcggcc gcctaactgt agtgccatat
     2401 ggaccagagt gtcaccagag taatggaata agccagcacc gccagcaagt agatccgaaa
     2461 cagcagcctg tcaaggacgt agcccactcg cagccagtcg gcagcgactt cgtccatctc
     2521 gtcctgcttc tccagaaaat gtcggatgct tgacagttcc tggagcagcc cccggacagc
     2581 gagagaggct tctcggggag gaggaagggg tgatccccgc ccccgtgggg tcttttccaa
     2641 atcttgtggt cccccaacat gggagcagtg attccccata gcaggcaaca gatcagaccc
     2701 actacaatcg tctgtcttgt tggcttgaaa ggttgcaggt ggccggtggg ccataggttg
     2761 ctcgccaaga cacagaatcc atgcaattct gtccagcaca aggtgtcgca accagtctgg
     2821 cacgggtctt tgcaggtcct gcttatgtac cagccgtaca atgaagatag tttccgccag
     2881 acttatcacc aggagtgcca tacacacgac gaaatacact cctatcagcg gagtgccgat
     2941 tgtggcagga agggtatcgg acacgatgat cagaaataca gaatagccga gcaacaaggt
     3001 aatcttaaag gagaccctct ccccgctatc aggtgggaga caaaatccca cgatgtccac
     3061 gaccatgaga aaaatactgg ggagcaacaa actaacggcg tagaacaatg gccttcttct
     3121 gatgataact gtgaatgtta cgtcagggta gggttccttg cagcattcat aaaatcgctc
     3181 gcttctttta ccggggattc ccacaaggtc ccactcccca ttcgggatgt aaccagagat
     3241 gtcagcctcc tgcatttgca gatccaggct ccatccgccg tagctccagc tcccgaattt
     3301 gagtttgcag tgctggacgt cgaagggaaa ccagcggacg tcgatatagc atgaggactt
     3361 gaagattcca gggggaaagt actggcagtg gcctgaagaa ttgaccaaca cgttggtgtg
     3421 aaatgttgca tcaaaccttt cgtctgcgct gttgaacagc aggatatctg gtttccagat
     3481 ttggccgtca gggaatctta cagtctttac acctgggtat tcgctcacat tccactgcaa
     3541 gtagtgatcg gtccaggaca tctgcaacca aatattagtg gtcagcacct ggttcttttc
     3601 atccacgtcc atgatctgca gcaaggagag gctgaagtac actgtcagtg gctgggaatc
     3661 atttgcgacc ggccgctcca gggggttata attcttaaca agctccttat acagtttgcg
     3721 ctggaactca ccctgcaggc taacgtgcag aagggaagca gccagggcga gccacacgcc
     3781 gcctggagaa cagcgcatgg tgaattcaag cttgctagct agtataactt cgtataggat
     3841 actttatacg aagttatcat tgggattctt cctattttga tccaagcatc accatcgacc
     3901 ctctagtcca gatctcacca tcgacccata acttcgtata gcatacatta tacgaagtta
     3961 tgtccctcga agaggttcga aacactgaaa agggcgaatt ctgcagatga tccactcgag
     4021 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     4081 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     4141 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     4201 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     4261 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     4321 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     4381 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     4441 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     4501 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     4561 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     4621 catcgatacc gtcgacccgg gcggccgctc gagcatgcat ctagagtcga cccgggcggc
     4681 cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     4741 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     4801 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     4861 gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata
     4921 aggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac
     4981 aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     5041 gccgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcattaatg
     5101 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5161 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5221 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5281 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5341 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5401 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5461 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5521 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5581 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5641 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5701 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5761 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5821 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5881 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5941 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6001 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6061 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6121 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6181 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6241 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6301 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6361 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6421 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6481 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6541 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6601 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6661 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6721 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6781 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6841 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6901 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6961 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7021 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     7081 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     7141 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     7201 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     7261 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     7321 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     7381 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     7441 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     7501 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gc
//
