LOCUS       sequence-45586-e        8673 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 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 8673)
  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 8673)
  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::PSAML141F,Y115F:5HT3HC-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..8673
                     /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(3238..3811,3812..3814,3815..3823)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3405..3422)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             join(3836..3838,3839..4552)
                     /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(3878..3899)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4139..4158)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4486..4507
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          4597..4729
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4712..4731
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    4738..5326
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4791..4811)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5209..5220)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5353..5370)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5359..5470
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5697..5716)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5762..5817
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5816..5835)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6176..6192)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6189..6211)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6200..6216
                     /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(6224..6230,6231..6248,6249..6254))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6269..6290
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6349..6544
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6395..6530
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6550..6684
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6587..6606)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6641..6660
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6752..6769)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6923..7511)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7003..7022)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7682..8473,8474..8542))
                     /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     8305..8324
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8543..8647)
                     /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 ggatatctcc ggacccgggc tagcaagctt gaattcacca tgcgctgttc
     1801 tccaggcggc gtgtggctcg ccctggctgc ttcccttctg cacgttagcc tgcagggtga
     1861 gttccagcgc aaactgtata aggagcttgt taagaattat aaccccctgg agcggccggt
     1921 cgcaaatgat tcccagccac tgacagtgta cttcagcctc tccttgctgc agatcatgga
     1981 cgtggatgaa aagaaccagg tgctgaccac taatatttgg ttgcagatgt cctggaccga
     2041 tcactacttg cagtggaatg tgagcgaata cccaggtgta aagactgtaa gattccctga
     2101 cggccaaatc tggaaaccag atatcctgct gttcaacagc gcagacgaaa ggtttgatgc
     2161 aacatttcac accaacgtgt tggtcaattc ttcaggccac tgccagtact tcccccctgg
     2221 aatcttcaag tcctcatgct atatcgacgt ccgctggttt cccttcgacg tccagcactg
     2281 caaactcaaa ttcgggagct ggagctacgg cggatggagc ctggatctgc aaatgcagga
     2341 ggctgacatc tctggttaca tcccgaatgg ggagtgggac cttgtgggaa tccccggtaa
     2401 aagaagcgag cgattttatg aatgctgcaa ggaaccctac cctgacgtaa cattcacagt
     2461 tatcatcaga agaaggccat tgttctacgc cgttagtttg ttgctcccca gtatttttct
     2521 catggtcgtg gacatcgtgg gattttgtct cccacctgat agcggggaga gggtctcctt
     2581 taagattacc ttgttgctcg gctattctgt atttctgatc atcgtgtccg atacccttcc
     2641 tgccacaatc ggcactccgc tgataggagt gtatttcgtc gtgtgtatgg cactcctggt
     2701 gataagtctg gcggaaacta tcttcattgt acggctggta cataagcagg acctgcaaag
     2761 acccgtgcca gactggttgc gacaccttgt gctggacaga attgcatgga ttctgtgtct
     2821 tggcgagcaa cctatggccc accggccacc tgcaaccttt caagccaaca agacagacga
     2881 ttgtagtggg tctgatctgt tgcctgctat ggggaatcac tgctcccatg ttgggggacc
     2941 acaagatttg gaaaagaccc cacgggggcg gggatcaccc cttcctcctc cccgagaagc
     3001 ctctctcgct gtccgggggc tgctccagga actgtcaagc atccgacatt ttctggagaa
     3061 gcaggacgag atggacgaag tcgctgccga ctggctgcga gtgggctacg tccttgacag
     3121 gctgctgttt cggatctact tgctggcggt gctggcttat tccattactc tggtgacact
     3181 ctggtccata tggcactaca gttaggcggc cgcaattccc cccgcccccc cccccccccc
     3241 ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     3301 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     3361 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     3421 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     3481 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     3541 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     3601 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc taagcgtatt caacaagggg
     3661 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     3721 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     3781 gtggttttcc tttgaaaaac acgatgataa tatggccaca accatggggg atccggtgag
     3841 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     3901 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     3961 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     4021 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     4081 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     4141 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     4201 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     4261 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     4321 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     4381 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     4441 cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     4501 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aatctagagt
     4561 cgaggtaccg atgaatagct aaggtcgagg ccgcaggtaa gtatcaaggt tacaagacag
     4621 gtttaaggag accaatagaa actgggcttg tcgagacaga gaagactctt gcgtttctga
     4681 taggcaccta ttggtcttac tgacatccac tttgcctttc tctccacagg tgtcgacaat
     4741 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     4801 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     4861 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     4921 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     4981 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     5041 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     5101 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     5161 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     5221 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     5281 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aattcgagct
     5341 cggtacgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     5401 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     5461 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     5521 acagcaaggg ggaggattgg gaagacaata gcaggccagc ttttgttccc tttagggtta
     5581 attgcgcgct tggcgctaat catggtcata gctgtttcca ctcctcaggt gcaggctgcc
     5641 tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt
     5701 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     5761 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     5821 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     5881 ttggcaacat atgccatatg ctggctgcca tgaacaaagg tggctataaa gaggtcatca
     5941 gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg
     6001 ttagattttt ttttatattt tgtttgtgtt atttttttct ttaacatccc taaaattttc
     6061 cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt
     6121 ccctcttctc ttatgaagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat
     6181 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6241 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6301 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta
     6361 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     6421 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6481 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     6541 caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6601 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6661 aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag
     6721 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6781 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6841 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6901 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6961 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     7021 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     7081 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     7141 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     7201 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7261 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7321 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     7381 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7441 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7501 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7561 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7621 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7681 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7741 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7801 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7861 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7921 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7981 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     8041 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     8101 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     8161 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     8221 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     8281 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8341 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8401 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8461 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8521 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8581 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8641 ttccgcgcac atttccccga aaagtgccac ctg
//
