LOCUS       sequence-373324-        7870 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of DRD2 with mEGFP tag on N-terminus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7870)
  AUTHORS   Foust DJ, Piston DW
  TITLE     Measuring G Protein Activation with Spectrally Resolved Fluorescence
            Fluctuation Spectroscopy
  JOURNAL   bioRxiv 2021.11.03.467169
REFERENCE   2  (bases 1 to 7870)
  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: "bioRxiv 
            2021.11.03.467169"
COMMENT     Sequence Label: pmEGFP-DRD2
FEATURES             Location/Qualifiers
     source          1..7870
                     /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"
     regulatory      1021..1030
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1027..1029,1030..1032,1033..1740)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(1072..1093)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1333..1352)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1680..1701
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3292..3309)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3298..3522
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3568..3996
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3655..3674)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3865..3886
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4005..4025)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4010..4339
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4190..4325
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4252..4271
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4388..5413
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4388..5413
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    5543..5664
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5580..5599)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5634..5653
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5713..5729)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5713..5729)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5726..5748)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5737..5753
                     /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(5761..5767,5768..5785,5786..5791))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5806..5827
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5944..5961)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6115..6703)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6195..6214)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6874..7665,7666..7734))
                     /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     7497..7516
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7735..7839)
                     /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 gctggctaga
      901 ctcgagggcc gcccagcttc caccatgggc ctccgggcgc tgatgctgtg gttgctggcg
      961 gcgacggggc tcgtgcgcga gtccctgcaa ggagagttcc aaaggaagct gtacccggtc
     1021 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     1081 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     1141 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     1201 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     1261 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     1321 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     1381 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     1441 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     1501 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     1561 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1621 aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
     1681 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1741 gatccactga atctgtcctg gtatgatgat gatctggaga ggcagaactg gagccggccc
     1801 ttcaacgggt cagacgggaa ggcggacaga ccccactaca actactatgc cacactgctc
     1861 accctgctca tcgctgtcat cgtcttcggc aacgtgctgg tgtgcatggc tgtgtctaga
     1921 gagaaggcgc tgcagaccac caccaactac ctgatcgtca gcctcgcagt ggccgacctc
     1981 ctcgtcgcca cactggtcat gccctgggtt gtctacctgg aggtggtagg tgagtggaaa
     2041 ttcagcagga ttcactgtga catcttcgtc actctggacg tcatgatgtg cacggcgagc
     2101 atcctgaact tgtgtgccat cagcatcgac aggtacacag ctgtggccat gcccatgctg
     2161 tacaatacgc gctacagctc caagcgccgg gtcaccgtca tgatctccat cgtctgggtc
     2221 ctgtccttca ccatctcctg cccactcctc ttcggactca ataacgcaga ccagaacgag
     2281 tgcatcattg ccaacccggc cttcgtggtc tactcctcca tcgtctcctt ctacgtgccc
     2341 ttcattgtca ccctgctggt ctacatcaag atctacattg tcctccgcag acgccgcaag
     2401 cgagtcaaca ccaaacgcag cagccgagct ttcagggccc acctgagggc tccactaaag
     2461 gaggctgccc ggcgagccca ggagctggag atggagatgc tctccagcac cagcccaccc
     2521 gagaggaccc ggtacagccc catcccaccc agccaccacc agctgactct ccccgacccg
     2581 tcccaccacg gtctccacag cactcctgac agccccgcca aaccagagaa gaatgggcat
     2641 gccaaagacc accccaagat tgccaagatc tttgagatcc agaccatgcc caatggcaaa
     2701 acccggacct ccctcaagac catgagccgt aggaagctct cccagcagaa ggagaagaaa
     2761 gccactcaga tgctcgccat tgttctcggc gtgttcatca tctgctggct gcccttcttc
     2821 atcacacaca tcctgaacat acactgtgac tgcaacatcc cgcctgtcct gtacagcgcc
     2881 ttcacgtggc tgggctatgt caacagcgcc gtgaacccca tcatctacac caccttcaac
     2941 attgagttcc gcaaggcctt cctgaagatc ctccactgct gactctgctg cctgcccgca
     3001 cagcagcctg cttcccacct ccctgcccag gccagccagc ctcacccttg cgaaccgtga
     3061 gcaggaaggc ctgggtggat cggcctcctc ttcaccccgg caggccctgc agtgttcgct
     3121 tggctccatg ctcctcactg cccgcacacc ctcactctgc cagggcagtg ctagtgagct
     3181 gggcatggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc agatatccag
     3241 cacagtggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg
     3301 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3361 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3421 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3481 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     3541 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     3601 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3661 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3721 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3781 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3841 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3901 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3961 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     4021 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4081 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4141 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4201 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4261 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     4321 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     4381 gcacgtgatg aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     4441 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     4501 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     4561 caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct
     4621 tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     4681 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
     4741 catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     4801 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     4861 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     4921 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     4981 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     5041 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     5101 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     5161 atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     5221 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     5281 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac
     5341 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     5401 ggcaaaggaa tagcacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt
     5461 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5521 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5581 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5641 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5701 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5761 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5821 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5881 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5941 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6001 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6061 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6121 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6181 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6241 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6301 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6361 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6421 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6481 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6541 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6601 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6661 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6721 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6781 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6841 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6901 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6961 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     7021 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7081 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7141 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7201 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7261 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7321 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7381 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7441 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7501 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7561 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7621 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7681 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7741 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7801 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7861 acctgacgtc
//
