LOCUS       sequence-45603-e        8553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PSAM-GlyR (L141F,Y115F) neuronal inhibitor, with IRES-EGFP
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8553)
  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 8553)
  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:GlyR-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..8553
                     /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(3118..3691,3692..3694,3695..3703)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3285..3302)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             join(3716..3718,3719..4432)
                     /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(3758..3779)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4019..4038)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4366..4387
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          4477..4609
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4592..4611
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    4618..5206
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4671..4691)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5089..5100)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5233..5250)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5239..5350
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5577..5596)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5642..5697
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5696..5715)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6056..6072)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6069..6091)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6080..6096
                     /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(6104..6110,6111..6128,6129..6134))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6149..6170
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6229..6424
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6275..6410
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6430..6564
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6467..6486)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6521..6540
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6632..6649)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6803..7391)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6883..6902)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7562..8353,8354..8422))
                     /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     8185..8204
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8423..8527)
                     /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 agagccctac ccagatgtca ccttcacagt
     2461 gaccatgcgg agacgcatgg gttattatct gatccaaatg tatatcccaa gcttgcttat
     2521 agtgattttg tcatggatct ccttctggat taatatggac gccgctccag ctagggtcgg
     2581 actgggcatc accacagtgc tgacaatgac tactcagagc tcaggcagcc gagccagctt
     2641 gcccaaggtt tcttacgtga aggccatcga tatctggatg gctgtctgcc ttctgtttgt
     2701 cttcagcgca ctgctggaat acgccgctgt caattttgtg tctcgacagc ataaagagct
     2761 gttgcggttc agaagaaaac gacgccacca caaagaggat gaggcaggag aaggacgctt
     2821 caactttagc gcctatggta tgggacctgc ttgcctccag gctaaagacg gaatttccgt
     2881 gaagggagcc aacaatagca acacaaccaa cccaccccct gctccatcta agagcccgga
     2941 ggaaatgcgc aaactcttta ttcagagagc gaaaaagatc gacaaaatct cccggatcgg
     3001 attccccatg gctttcctga ttttcaacat gttttattgg atcatctaca agattgtgcg
     3061 aagggaggac gtacacaacc agtaagcggc cgcaattccc cccgcccccc cccccccccc
     3121 ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     3181 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     3241 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     3301 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     3361 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     3421 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     3481 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc taagcgtatt caacaagggg
     3541 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     3601 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     3661 gtggttttcc tttgaaaaac acgatgataa tatggccaca accatggggg atccggtgag
     3721 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     3781 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     3841 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     3901 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     3961 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     4021 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     4081 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     4141 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     4201 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     4261 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     4321 cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     4381 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aatctagagt
     4441 cgaggtaccg atgaatagct aaggtcgagg ccgcaggtaa gtatcaaggt tacaagacag
     4501 gtttaaggag accaatagaa actgggcttg tcgagacaga gaagactctt gcgtttctga
     4561 taggcaccta ttggtcttac tgacatccac tttgcctttc tctccacagg tgtcgacaat
     4621 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     4681 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     4741 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     4801 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     4861 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     4921 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     4981 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     5041 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     5101 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     5161 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aattcgagct
     5221 cggtacgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     5281 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     5341 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     5401 acagcaaggg ggaggattgg gaagacaata gcaggccagc ttttgttccc tttagggtta
     5461 attgcgcgct tggcgctaat catggtcata gctgtttcca ctcctcaggt gcaggctgcc
     5521 tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt
     5581 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     5641 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     5701 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     5761 ttggcaacat atgccatatg ctggctgcca tgaacaaagg tggctataaa gaggtcatca
     5821 gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg
     5881 ttagattttt ttttatattt tgtttgtgtt atttttttct ttaacatccc taaaattttc
     5941 cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt
     6001 ccctcttctc ttatgaagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat
     6061 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6121 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6181 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta
     6241 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     6301 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6361 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     6421 caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6481 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6541 aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag
     6601 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6661 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6721 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6781 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6841 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6901 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6961 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     7021 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     7081 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7141 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7201 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     7261 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7321 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7381 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7441 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7501 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7561 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7621 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7681 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7741 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7801 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7861 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7921 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7981 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     8041 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     8101 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     8161 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8221 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8281 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8341 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8401 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8461 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8521 ttccgcgcac atttccccga aaagtgccac ctg
//
