LOCUS       sequence-63773-e        6832 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses hM3D (Gq) neuronal activator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6832)
  AUTHORS   Armbruster BN, Li X, Pausch MH, Herlitze S, Roth BL
  TITLE     Evolving the lock to fit the key to create a family of G 
            protein-coupled receptors potently activated by an inert ligand.
  JOURNAL   Proc Natl Acad Sci U S A. 2007 Mar 20;104(12):5163-8. Epub 2007 Mar 
            2.
  PUBMED    17360345
REFERENCE   2  (bases 1 to 6832)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2007-03-20"; volume: "104(12)"; pages: 
            "5163-8. Epub 2007 Mar 2"
COMMENT     Sequence Label: pcDNA5/FRT-HA-hM3D(Gq)
FEATURES             Location/Qualifiers
     source          1..6832
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             941..967
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2785..2802)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2791..3015
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3148..3167)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3298..3345
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3353..4375
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3353..4375
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4505..4626
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4542..4561)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4596..4615
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4675..4691)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4675..4691)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4688..4710)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4699..4715
                     /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(4723..4729,4730..4747,4748..4753))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4768..4789
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4906..4923)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5077..5665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5157..5176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5836..6627,6628..6696))
                     /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     6459..6478
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6697..6801)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccaccatg tacccatacg atgttccaga
      961 ttacgctatg accttgcaca ataacagtac aacctcgcct ttgtttccaa acatcagctc
     1021 ctcctggata cacagcccct ccgatgcagg gctgcccccg ggaaccgtca ctcatttcgg
     1081 cagctacaat gtttctcgag cagctggcaa tttctcctct ccagacggta ccaccgatga
     1141 ccctctggga ggtcataccg tctggcaagt ggtcttcatc gctttcttaa cgggcatcct
     1201 ggccttggtg accatcatcg gcaacatcct ggtaattgtg tcatttaagg tcaacaagca
     1261 gctgaagacg gtcaacaact acttcctctt aagcctggcc tgtgccgatc tgattatcgg
     1321 ggtcatttca atgaatctgt ttacgaccta catcatcatg aatcgatggg ccttagggaa
     1381 cttggcctgt gacctctggc ttgccattga ctgcgtagcc agcaatgcct ctgttatgaa
     1441 tcttctggtc atcagctttg acagatactt ttccatcacg aggccgctca cgtaccgagc
     1501 caaacgaaca acaaagagag ccggtgtgat gatcggtctg gcttgggtca tctcctttgt
     1561 cctttgggct cctgccatct tgttctggca atactttgtt ggaaagagaa ctgtgcctcc
     1621 gggagagtgc ttcattcagt tcctcagtga gcccaccatt acttttggca cagccatcgc
     1681 tggtttttat atgcctgtca ccattatgac tattttatac tggaggatct ataaggaaac
     1741 tgaaaagcgt accaaagagc ttgctggcct gcaagcctct gggacagagg cagagacaga
     1801 aaactttgtc caccccacgg gcagttctcg aagctgcagc agttacgaac ttcaacagca
     1861 aagcatgaaa cgctccaaca ggaggaagta tggccgctgc cacttctggt tcacaaccaa
     1921 gagctggaaa cccagctccg agcagatgga ccaagaccac agcagcagtg acagttggaa
     1981 caacaatgat gctgctgcct ccctggagaa ctccgcctcc tccgacgagg aggacattgg
     2041 ctccgagacg agagccatct actccatcgt gctcaagctt ccgggtcaca gcaccatcct
     2101 caactccacc aagttaccct catcggacaa cctgcaggtg cctgaggagg agctggggat
     2161 ggtggacttg gagaggaaag ccgacaagct gcaggcccag aagagcgtgg acgatggagg
     2221 cagttttcca aaaagcttct ccaagcttcc catccagcta gagtcagccg tggacacagc
     2281 taagacttct gacgtcaact cctcagtggg taagagcacg gccactctac ctctgtcctt
     2341 caaggaagcc actctggcca agaggtttgc tctgaagacc agaagtcaga tcactaagcg
     2401 gaaaaggatg tccctggtca aggagaagaa agcggcccag accctcagtg cgatcttgct
     2461 tgccttcatc atcacttgga ccccatacaa catcatggtt ctggtgaaca ccttttgtga
     2521 cagctgcata cccaaaacct tttggaatct gggctactgg ctgtgctaca tcaacagcac
     2581 cgtgaacccc gtgtgctatg ctctgtgcaa caaaacattc agaaccactt tcaagatgct
     2641 gctgctgtgc cagtgtgaca aaaaaaagag gcgcaagcag cagtaccagc agagacagtc
     2701 ggtcattttt cacaagcgcg cacccgagca ggccttgtag gcggccgctc gagtctagag
     2761 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     2821 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2881 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2941 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     3001 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     3061 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3121 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3181 cgttcgccgg ctttccccgt caagctctaa atcgggggtc cctttagggt tccgatttag
     3241 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtacctagaa
     3301 gttcctattc cgaagttcct attctctaga aagtatagga acttccttgg ccaaaaagcc
     3361 tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga
     3421 cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg
     3481 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta
     3541 tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag
     3601 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc
     3661 tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc
     3721 ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata
     3781 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac
     3841 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg
     3901 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
     3961 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
     4021 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
     4081 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc
     4141 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
     4201 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
     4261 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt
     4321 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagcacgt
     4381 actacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4441 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4501 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4561 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4621 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4681 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4741 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4801 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4861 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4921 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4981 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5041 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5101 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5161 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5221 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5281 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5341 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5401 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5461 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     5521 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5581 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5641 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5701 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5761 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5821 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5881 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5941 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6001 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6061 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6121 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6181 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6241 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6301 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6361 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6421 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6481 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6541 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6601 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6661 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6721 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6781 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
