LOCUS       sequence-177075-        7337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses DRD2-iTango component in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7337)
  AUTHORS   Lee D, Creed M, Jung K, Stefanelli T, Wendler DJ, Oh WC, Mignocchi 
            NL, Luscher C, Kwon HB
  TITLE     Temporally precise labeling and control of neuromodulatory circuits 
            in the mammalian brain.
  JOURNAL   Nat Methods. 2017 May;14(5):495-503. doi: 10.1038/nmeth.4234. Epub 
            2017 Apr 3.
  PUBMED    28369042
REFERENCE   2  (bases 1 to 7337)
  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"; doi: 
            "10.1038/nmeth.4234"; journalName: "Nat Methods"; date: "2017-05"; 
            volume: "14"; issue: "5"; pages: "495-503"
COMMENT     Sequence Label: pCMV-DRD2-V2tail-TevN-BLITz1-TetR-VP16
FEATURES             Location/Qualifiers
     source          1..7337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             2470..2496
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             3310..3330
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(3331..3948,3949..4335)
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus 
                     VP16"
                     /label=tTA
                     /note="In the Tet-Off(R) system, tTA binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="SRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVK
                     NKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRP
                     TEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTD
                     SMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTIE
                     GLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHAD
                     ALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(3401..3420)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(4344..4363)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4345..4363)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    4368..4502
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4392..4411)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4446..4465
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4615..4635)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4615..4633)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4654..4670)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4654..4670)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4667..4689)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4678..4694
                     /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(4702..4708,4709..4726,4727..4732))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4747..4768
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4885..4902)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5055..5643)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5135..5154)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5814..6605,6606..6674))
                     /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     6437..6456
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6675..6779)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6805..7260)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6942..6963)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7154..7173
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaatga agacgatcat cgccctgagc
     1081 tacatcttct gcctggtatt cgccgactac aaggacgatg atgacgccag catggatcca
     1141 ctgaacctgt cctggtatga tgatgatctg gagaggcaga actggagccg gcccttcaat
     1201 gggtcagaag ggaaggcaga caggccccac tacaactact atgccatgct gctcaccctc
     1261 ctcatcttta tcatcgtctt tggcaatgtg ctggtgtgca tggctgtatc ccgagagaag
     1321 gctttgcaga ccaccaccaa ctacttgata gtcagccttg ctgtggctga tcttctggtg
     1381 gccacactgg taatgccgtg ggttgtctac ctggaggtgg tgggtgagtg gaaattcagc
     1441 aggattcact gtgacatctt tgtcactctg gatgtcatga tgtgcacagc aagcatcctg
     1501 aacctgtgtg ccatcagcat tgacaggtac acagctgtgg caatgcccat gctgtataac
     1561 acacgctaca gctccaagcg ccgagttact gtcatgattg ccattgtctg ggtcctgtcc
     1621 ttcaccatct cctgcccact gctcttcgga ctcaacaata cagaccagaa tgagtgtatc
     1681 attgccaacc ctgcctttgt ggtctactcc tccattgtct cattctacgt gcccttcatc
     1741 gtcactctgc tggtctatat caaaatctac atcgtcctcc ggaagcgccg gaagcgggtc
     1801 aacaccaagc gcagcagtcg agctttcaga gccaacctga agacaccact caaggatgct
     1861 gcccgccgag ctcaggagct ggaaatggag atgctgtcaa gcaccagccc cccagagagg
     1921 acccggtata gccccatccc tcccagtcac caccagctca ctctccctga tccatcccac
     1981 cacggcctac atagcaaccc tgacagtcct gccaaaccag agaagaatgg gcacgccaag
     2041 attgtcaatc ccaggattgc caagttcttt gagatccaga ccatgcccaa tggcaaaacc
     2101 cggacctccc ttaagacgat gagccgcaga aagctctccc agcagaagga gaagaaagcc
     2161 actcagatgc ttgccattgt tctcggtgtg ttcatcatct gctggctgcc cttcttcatc
     2221 acgcacatcc tgaatataca ctgtgattgc aacatcccac cagtcctcta cagcgccttc
     2281 acatggctgg gctatgtcaa cagtgccgtc aaccccatca tctacaccac cttcaacatc
     2341 gagttccgca aggccttcat gaagatcttg cactgcgaat tcggacgcac cccacccagc
     2401 ctgggtcccc aagatgagtc ctgcaccacc gccagctcct ccctggccaa ggacacttca
     2461 tcgcttcaac ttcctcctct tgaacgtctt actctcggtg gctctggagg tctcgagatg
     2521 ggcgagagcc tgttcaaggg acctagggac tacaacccta tcagtagcac aatttgtcac
     2581 ctgaccaacg agagtgatgg ccacacaaca agcctgtacg gcatcgggtt cggacccttt
     2641 atcatcacca acaagcacct gttcaggcgg aataatggca ctctgctggt gcagagcctg
     2701 cacggggtgt tcaaagtgaa gaacacaacc actctgcagc agcacctgat cgatgggcgg
     2761 gatatgatca tcattaggat gcccaaggac ttcccccctt ttcctcagaa actgaagttc
     2821 cgagagcccc agagagagga gagaatctgt ctggtgacca caaactttca ggagctcggt
     2881 agtggtagtg gggagtttct ggcaaccaca ctggaacgga tcgagaaaaa tttcgtgatt
     2941 actgatccga gactgcctga caacccaatc atttttgcga gcgattcctt cctgcagctg
     3001 acagaatatt ctcgggaaga gatcctgggg cgcaattgcc gttttctgca gggacccgag
     3061 acagaccgtg ccactgttcg gaaaatcaga gatgctattg acaaccagac tgaagtgacc
     3121 gttcagctga tcaattatac caagagcggc aagaagttct ggaacgtgtt ccacctgcag
     3181 ccgatgcgcg attataaggg cgacgtccag tacttcattg gcgtgcagct ggatggcacc
     3241 gaacgtcttc atggcgccgc tgagcgtgag gcggtctgcc tgatcaaaaa gacagccttt
     3301 cagattgctg agaacctgta cttccagggc tctagattag ataaaagtaa agtgattaac
     3361 agcgcattag agctgcttaa tgaggtcgga atcgaaggtt taacaacccg taaactcgcc
     3421 cagaagctag gtgtagagca gcctacattg tattggcatg taaaaaataa gcgggctttg
     3481 ctcgacgcct tagccattga gatgttagat aggcaccata ctcacttttg ccctttagaa
     3541 ggggaaagct ggcaagattt tttacgtaat aacgctaaaa gttttagatg tgctttacta
     3601 agtcatcgcg atggagcaaa agtacattta ggtacacggc ctacagaaaa acagtatgaa
     3661 actctcgaaa atcaattagc ctttttatgc caacaaggtt tttcactaga gaatgcatta
     3721 tatgcactca gcgctgtggg gcattttact ttaggttgcg tattggaaga tcaagagcat
     3781 caagtcgcta aagaagaaag ggaaacacct actactgata gtatgccgcc attattacga
     3841 caagctatcg aattatttga tcaccaaggt gcagagccag ccttcttatt cggccttgaa
     3901 ttgatcatat gcggattaga aaaacaactt aaatgtgaaa gtgggtccgc gtacagccgc
     3961 gcgcgtacga aaaacaatta cgggtctacc atcgagggcc tgctcgatct cccggacgac
     4021 gacgcccccg aagaggcggg gctggcggct ccgcgcctgt cctttctccc cgcgggacac
     4081 acgcgcagac tgtcgacggc ccccccgacc gatgtcagcc tgggggacga gctccactta
     4141 gacggcgagg acgtggcgat ggcgcatgcc gacgcgctag acgatttcga tctggacatg
     4201 ttgggggacg gggattcccc gggtccggga tttacccccc acgactccgc cccctacggc
     4261 gctctggata tggccgactt cgagtttgag cagatgttta ccgatgccct tggaattgac
     4321 gagtacggtg ggtaggctag cctccctata gtgagtcgta ttacgtagat ccagacatga
     4381 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     4441 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     4501 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     4561 tttaattcgc gtccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta
     4621 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4681 ttatccgctc acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg
     4741 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4801 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4861 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4921 gcgggagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4981 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5041 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5101 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5161 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5221 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5281 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5341 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5401 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5461 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5521 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5581 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5641 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5701 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5761 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5821 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5881 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5941 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6001 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6061 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6121 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6181 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6241 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6301 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6361 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6421 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6481 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6541 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6601 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6661 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6721 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6781 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     6841 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     6901 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     6961 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     7021 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     7081 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     7141 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     7201 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     7261 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     7321 tattacgcca gtcgacc
//
