LOCUS       sequence-45581-e        8698 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PSAM-5HT3 HC (Q79G,Q139G) neuronal activator, with 
            IRES-EGFP marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8698)
  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 8698)
  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: CAG::PSAMQ79G,Q139G:5HT3HC-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..8698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     misc_feature    join(3262..3835,3836..3838,3839..3847)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3429..3446)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             join(3860..3862,3863..4576)
                     /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(3902..3923)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4163..4182)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4510..4531
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          4621..4753
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4736..4755
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    4762..5350
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4815..4835)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5233..5244)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5377..5394)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5383..5494
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5721..5740)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5786..5841
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5840..5859)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6200..6216)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6213..6235)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6224..6240
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6248..6254,6255..6272,6273..6278))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6293..6314
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6373..6568
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6419..6554
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6574..6708
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6611..6630)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6665..6684
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6776..6793)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6947..7535)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7027..7046)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7706..8497,8498..8566))
                     /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     8329..8348
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8567..8671)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg ggcgagggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tgttgtgctg tctcatcatt ttggcaaaga attcggtacc gcgggcccgg
     1741 cgcgccagct ggatagcttg aattcaccat gcgctgttct ccaggcggcg tgtggctcgc
     1801 cctggctgct tcccttctgc acgttagcct gcagggtgag ttccagcgca aactgtataa
     1861 ggagcttgtt aagaattata accccctgga gcggccggtc gcaaatgatt cccagccact
     1921 gacagtgtac ttcagcctct ccttgctgca gatcatggac gtggatgaaa agaaccaggt
     1981 gctgaccact aatatttggt tgggcatgtc ctggaccgat cactacttgc agtggaatgt
     2041 gagcgaatac ccaggtgtaa agactgtaag attccctgac ggccaaatct ggaaaccaga
     2101 tatcctgctg tacaacagcg cagacgaaag gtttgatgca acatttcaca ccaacgtgtt
     2161 ggtcaattct tcaggccact gcggctacct gccccctgga atcttcaagt cctcatgcta
     2221 tatcgacgtc cgctggtttc ccttcgacgt ccagcactgc aaactcaaat tcgggagctg
     2281 gagctacggc ggatggagcc tggatctgca aatgcaggag gctgacatct ctggttacat
     2341 cccgaatggg gagtgggacc ttgtgggaat ccccggtaaa agaagcgagc gattttatga
     2401 atgctgcaag gaaccctacc ctgacgtaac attcacagtt atcatcagaa gaaggccatt
     2461 gttctacgcc gttagtttgt tgctccccag tatttttctc atggtcgtgg acatcgtggg
     2521 attttgtctc ccacctgata gcggggagag ggtctccttt aagattacct tgttgctcgg
     2581 ctattctgta tttctgatca tcgtgtccga tacccttcct gccacaatcg gcactccgct
     2641 gataggagtg tatttcgtcg tgtgtatggc actcctggtg ataagtctgg cggaaactat
     2701 cttcattgta cggctggtac ataagcagga cctgcaaaga cccgtgccag actggttgcg
     2761 acaccttgtg ctggacagaa ttgcatggat tctgtgtctt ggcgagcaac ctatggccca
     2821 ccggccacct gcaacctttc aagccaacaa gacagacgat tgtagtgggt ctgatctgtt
     2881 gcctgctatg gggaatcact gctcccatgt tgggggacca caagatttgg aaaagacccc
     2941 acgggggcgg ggatcacccc ttcctcctcc ccgagaagcc tctctcgctg tccgggggct
     3001 gctccaggaa ctgtcaagca tccgacattt tctggagaag caggacgaga tggacgaagt
     3061 cgctgccgac tggctgcgag tgggctacgt ccttgacagg ctgctgtttc ggatctactt
     3121 gctggcggtg ctggcttatt ccattactct ggtgacactc tggtccatat ggcactacag
     3181 ttaggcggcc gcctcgagtc tagatctccg gacccgggct agcgcaattg cggccgcaat
     3241 tccccccgcc cccccccccc ccccctctcc ctcccccccc cctaacgtta ctggccgaag
     3301 ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc
     3361 ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg
     3421 tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc
     3481 tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc
     3541 cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa
     3601 ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct
     3661 ctcctaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     3721 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
     3781 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc
     3841 cacaaccatg ggggatccgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     3901 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     3961 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     4021 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     4081 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     4141 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     4201 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     4261 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     4321 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     4381 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     4441 gctgcccgac aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa
     4501 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     4561 cgagctgtac aagtaatcta gagtcgaggt accgatgaat agctaaggtc gaggccgcag
     4621 gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga
     4681 cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc
     4741 tttctctcca caggtgtcga caatcaacct ctggattaca aaatttgtga aagattgact
     4801 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     4861 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     4921 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     4981 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     5041 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     5101 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     5161 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     5221 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     5281 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     5341 gcctccccgc ctggaattcg agctcggtac gatcagcctc gactgtgcct tctagttgcc
     5401 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     5461 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     5521 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     5581 cagcttttgt tccctttagg gttaattgcg cgcttggcgc taatcatggt catagctgtt
     5641 tccactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg
     5701 gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag
     5761 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     5821 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
     5881 cagaatgagt atttggttta gagtttggca acatatgcca tatgctggct gccatgaaca
     5941 aaggtggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt
     6001 ccatagaaaa gccttgactt gaggttagat ttttttttat attttgtttg tgttattttt
     6061 ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc
     6121 ctgactactc ccagtcatag ctgtccctct tctcttatga agatccctcg acctgcagcc
     6181 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6241 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6301 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6361 gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     6421 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     6481 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     6541 cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt
     6601 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     6661 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat
     6721 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     6781 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6841 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6901 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6961 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     7021 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     7081 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     7141 aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7201 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7261 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7321 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     7381 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7441 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7501 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     7561 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     7621 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7681 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7741 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7801 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7861 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7921 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7981 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     8041 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     8101 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     8161 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     8221 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     8281 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     8341 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     8401 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     8461 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     8521 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     8581 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     8641 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgc
//
