LOCUS       sequence-244565-        6497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SPARK DRD1 transmembrane component in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6497)
  AUTHORS   Kim CK, Cho KF, Kim MW, Ting AY
  TITLE     Luciferase-LOV BRET enables versatile and specific transcriptional 
            readout of cellular protein-protein interactions.
  JOURNAL   eLife. 2019 Apr 3;8. pii: 43826. doi: 10.7554/eLife.43826.
  PUBMED    30942168
REFERENCE   2  (bases 1 to 6497)
  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: "eLife."; 
            date: "2019-04-3"; volume: "8. pii: 43826. doi"; pages: " 
            10.7554/eLife.43826"
COMMENT     Sequence Label: pAAV-DRD1-NNES-eLOV-TEVcs-Flag-Gal4-V5
FEATURES             Location/Qualifiers
     source          1..6497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   6..135
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        206..509
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        510..713
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     664..684
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     758..777
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        758..776
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2611..2631
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
     CDS             2644..2667
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2668..3105
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="KLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTKR
                     SPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNKD
                     AVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             3106..3447
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     CDS             3448..3489
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3523..3540)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3529..3736
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3765..3905
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3765..3894
                     /label=AAV2 ITR
     rep_origin      3980..4435
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4067..4086)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4277..4298
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4452..4471)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4571..4593
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4631..4649)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4717..4821
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4822..4890,4891..5682)
                     /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     complement(5040..5059)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5853..6441
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6342..6361
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
       61 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
      121 tcactagggg ttcctgcggc cgcacgcgtt gtacatagta atcaattacg gggtcattag
      181 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      241 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      301 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      361 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      421 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      481 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      541 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      601 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      661 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      721 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      781 cccaagcttg ccaccatgcg cacattgaat acttcagcga tggacggaac gggcctcgtg
      841 gtcgaacggg actttagcgt tagaattctt accgcttgct tcctgtcact gcttatcctg
      901 tctactctcc tgggtaatac acttgtgtgc gccgccgtga tcagattccg gcacttgcgg
      961 tccaaagtga ctaacttttt tgtcatctca ctggccgtct ctgatctgct ggtggctgtc
     1021 ctcgtgatgc catggaaagc agtggccgag atcgccgggt tctggccctt cgggtcattt
     1081 tgcaatatct gggtggcatt cgacatcatg tgtagcacgg cctcaattct caatctctgt
     1141 gtcattagtg tcgataggta ttgggccatc agctctccct tccgatacga gcggaagatg
     1201 accccaaagg ccgcctttat actgatatca gtcgcctgga ccctcagcgt gctgatttct
     1261 ttcatccctg tgcaattgtc ctggcacaag gctaaaccaa cgagtcccag cgacggaaac
     1321 gccactagtc tggccgagac tatcgacaat tgtgacagta gcctgtccag gacctacgct
     1381 atcagcagtt cagtgatctc cttttacatt cctgtcgcga ttatgatcgt gacctacaca
     1441 aggatctata gaatcgcaca gaagcaaatc cggaggatcg cggctttgga gagagccgct
     1501 gtgcacgcaa agaattgtca gacgacgacc ggcaacggta aacccgtcga gtgttcccag
     1561 ccagagtcca gcttcaaaat gtcatttaag cgcgaaacta aggttctgaa gaccctctct
     1621 gtgattatgg gcgtgtttgt gtgttgctgg ctccccttct tcattttgaa ttgtatactg
     1681 cctttctgcg gcagtggcga gactcagcct ttctgtatag actcaaacac ttttgacgtc
     1741 ttcgtttggt tcggttgggc caattcctcc cttaatccta ttatatacgc atttaatgct
     1801 gattttcgca aagcattctc caccctgctg ggctgttacc ggctttgtcc cgccaccaat
     1861 aacgcaattg agaccgtatc tattaacaat aacggcgcag ctatgttctc ctctcaccat
     1921 gagccaagag gttcaattag caaagagtgt aatcttgttt acttgatacc tcacgcagtt
     1981 gggagttcag aagaccttaa gaaagaagaa gccgccggca tcgctaggcc cctcgaaaaa
     2041 ctgagccctg cactttctgt gatcctggat tatgacactg atgtgagcct ggagaaaatt
     2101 caaccaataa cacagaacgg tcaacatcca actgagctgg ccgagaagct ggccggcctg
     2161 gacatcaacg gtggagctag cggcagccgg gcgaccaccc tggaacgtat tgaaaagagc
     2221 tttgtgatta ccgatccgcg tctgccggat aacccgatta tttttgtgag cgatagcttt
     2281 ctgcagctga ccgaatatag ccgtgaagaa attctgggcc gtaactgccg ttttctgcag
     2341 ggcccggaaa ccgatcgtgc gaccgtgcgt aaaattcgtg atgcgattga taaccagacc
     2401 gaagtgaccg tgcagctgat taactatacc aaaagcggca aaaaattttg gaacctgttt
     2461 catctgcagc cgatgcgtga tcagaaaggc gatgtgcagt attttattgg cgtgcagctg
     2521 gatggtaccg aacgtgtgcg tgatgcggcg gaacgtgaag ctgtgatgct ggtcaagaaa
     2581 accgcggagg aaatcgacga agccgctaag gaaaatctct acttccagat gggcggagga
     2641 tccgactaca aggatgacga cgataagaag ctactgtctt ctatcgaaca agcatgcgat
     2701 atttgccgac ttaaaaagct caagtgctcc aaagaaaaac cgaagtgcgc caagtgtctg
     2761 aagaacaact gggagtgtcg ctactctccc aaaaccaaaa ggtctccgct gactagggca
     2821 catctgacag aagtggaatc aaggctagaa agactggaac agctatttct actgattttt
     2881 cctcgagaag accttgacat gattttgaaa atggattctt tacaggatat aaaagcattg
     2941 ttaacaggat tatttgtaca agataatgtg aataaagatg ccgtcacaga tagattggct
     3001 tcagtggaga ctgatatgcc tctaacattg agacagcata gaataagtgc gacatcatca
     3061 tcggaagaga gtagtaacaa aggtcaaaga cagttgactg tatcggccaa ttttaatcaa
     3121 agtgggaata ttgctgatag ctcattgtcc ttcactttca ctaacagtag caacggtccg
     3181 aacctcataa caactcaaac aaattctcaa gcgctttcac aaccaattgc ctcctctaac
     3241 gttcatgata acttcatgaa taatgaaatc acggctagta aaattgatga tggtaataat
     3301 tcaaaaccac tgtcacctgg ttggacggac caaactgcgt ataacgcgtt tggaatcact
     3361 acagggatgt ttaataccac tacaatggat gatgtatata actatctatt cgatgatgaa
     3421 gataccccac caaacccaaa aaaagagggc aagccgatcc ctaaccctct gctgggcctg
     3481 gacagcactt aagaattcct cgagctagag ctcgctgatc agcctcgact gtgccttcta
     3541 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     3601 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     3661 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     3721 gcaggcatgc tggggaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     3781 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     3841 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     3901 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3961 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4021 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     4081 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4141 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     4201 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4261 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     4321 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     4381 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     4441 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4501 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4561 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4621 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4681 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4741 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4801 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4861 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4921 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4981 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5041 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5101 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5161 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5221 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5281 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5341 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5401 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5461 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5521 ggagccggtg agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc
     5581 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5641 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5701 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5761 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5821 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5881 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5941 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6001 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6061 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6121 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6181 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6241 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6301 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6361 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6421 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6481 tgctggcctt ttgctca
//
