LOCUS       sequence-177078-        7021 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses DRD2-iTango component in mammalian cells for AAV 
            production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7021)
  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 7021)
  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: pAAV-hSYN-DRD2-V2tail-TevN-BLITz1-TetR-VP16-bGHpA
FEATURES             Location/Qualifiers
     source          1..7021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1426..1452
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             2266..2286
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(2287..2904,2905..3291)
                     /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(2357..2376)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     polyA_signal    3313..3520
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3690..3819
                     /label=AAV2 ITR
     rep_origin      complement(4056..4644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4136..4155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4815..5606,5607..5675))
                     /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     5438..5457
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5676..5780)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5806..6261)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5943..5964)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6155..6174
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6333..6462
                     /label=AAV2 ITR
     promoter        6562..7009
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 aagcttgcca ccatgaagac gatcatcgcc ctgagctaca tcttctgcct ggtattcgcc
       61 gactacaagg acgatgatga cgccagcatg gatccactga acctgtcctg gtatgatgat
      121 gatctggaga ggcagaactg gagccggccc ttcaatgggt cagaagggaa ggcagacagg
      181 ccccactaca actactatgc catgctgctc accctcctca tctttatcat cgtctttggc
      241 aatgtgctgg tgtgcatggc tgtatcccga gagaaggctt tgcagaccac caccaactac
      301 ttgatagtca gccttgctgt ggctgatctt ctggtggcca cactggtaat gccgtgggtt
      361 gtctacctgg aggtggtggg tgagtggaaa ttcagcagga ttcactgtga catctttgtc
      421 actctggatg tcatgatgtg cacagcaagc atcctgaacc tgtgtgccat cagcattgac
      481 aggtacacag ctgtggcaat gcccatgctg tataacacac gctacagctc caagcgccga
      541 gttactgtca tgattgccat tgtctgggtc ctgtccttca ccatctcctg cccactgctc
      601 ttcggactca acaatacaga ccagaatgag tgtatcattg ccaaccctgc ctttgtggtc
      661 tactcctcca ttgtctcatt ctacgtgccc ttcatcgtca ctctgctggt ctatatcaaa
      721 atctacatcg tcctccggaa gcgccggaag cgggtcaaca ccaagcgcag cagtcgagct
      781 ttcagagcca acctgaagac accactcaag gatgctgccc gccgagctca ggagctggaa
      841 atggagatgc tgtcaagcac cagcccccca gagaggaccc ggtatagccc catccctccc
      901 agtcaccacc agctcactct ccctgatcca tcccaccacg gcctacatag caaccctgac
      961 agtcctgcca aaccagagaa gaatgggcac gccaagattg tcaatcccag gattgccaag
     1021 ttctttgaga tccagaccat gcccaatggc aaaacccgga cctcccttaa gacgatgagc
     1081 cgcagaaagc tctcccagca gaaggagaag aaagccactc agatgcttgc cattgttctc
     1141 ggtgtgttca tcatctgctg gctgcccttc ttcatcacgc acatcctgaa tatacactgt
     1201 gattgcaaca tcccaccagt cctctacagc gccttcacat ggctgggcta tgtcaacagt
     1261 gccgtcaacc ccatcatcta caccaccttc aacatcgagt tccgcaaggc cttcatgaag
     1321 atcttgcact gcgaattcgg acgcacccca cccagcctgg gtccccaaga tgagtcctgc
     1381 accaccgcca gctcctccct ggccaaggac acttcatcgc ttcaacttcc tcctcttgaa
     1441 cgtcttactc tcggtggctc tggaggtctc gagatgggcg agagcctgtt caagggacct
     1501 agggactaca accctatcag tagcacaatt tgtcacctga ccaacgagag tgatggccac
     1561 acaacaagcc tgtacggcat cgggttcgga ccctttatca tcaccaacaa gcacctgttc
     1621 aggcggaata atggcactct gctggtgcag agcctgcacg gggtgttcaa agtgaagaac
     1681 acaaccactc tgcagcagca cctgatcgat gggcgggata tgatcatcat taggatgccc
     1741 aaggacttcc ccccttttcc tcagaaactg aagttccgag agccccagag agaggagaga
     1801 atctgtctgg tgaccacaaa ctttcaggag ctcggtagtg gtagtgggga gtttctggca
     1861 accacactgg aacggatcga gaaaaatttc gtgattactg atccgagact gcctgacaac
     1921 ccaatcattt ttgcgagcga ttccttcctg cagctgacag aatattctcg ggaagagatc
     1981 ctggggcgca attgccgttt tctgcaggga cccgagacag accgtgccac tgttcggaaa
     2041 atcagagatg ctattgacaa ccagactgaa gtgaccgttc agctgatcaa ttataccaag
     2101 agcggcaaga agttctggaa cgtgttccac ctgcagccga tgcgcgatta taagggcgac
     2161 gtccagtact tcattggcgt gcagctggat ggcaccgaac gtcttcatgg cgccgctgag
     2221 cgtgaggcgg tctgcctgat caaaaagaca gcctttcaga ttgctgagaa cctgtacttc
     2281 cagggctcta gattagataa aagtaaagtg attaacagcg cattagagct gcttaatgag
     2341 gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt agagcagcct
     2401 acattgtatt ggcatgtaaa aaataagcgg gctttgctcg acgccttagc cattgagatg
     2461 ttagataggc accatactca cttttgccct ttagaagggg aaagctggca agatttttta
     2521 cgtaataacg ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg agcaaaagta
     2581 catttaggta cacggcctac agaaaaacag tatgaaactc tcgaaaatca attagccttt
     2641 ttatgccaac aaggtttttc actagagaat gcattatatg cactcagcgc tgtggggcat
     2701 tttactttag gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa
     2761 acacctacta ctgatagtat gccgccatta ttacgacaag ctatcgaatt atttgatcac
     2821 caaggtgcag agccagcctt cttattcggc cttgaattga tcatatgcgg attagaaaaa
     2881 caacttaaat gtgaaagtgg gtccgcgtac agccgcgcgc gtacgaaaaa caattacggg
     2941 tctaccatcg agggcctgct cgatctcccg gacgacgacg cccccgaaga ggcggggctg
     3001 gcggctccgc gcctgtcctt tctccccgcg ggacacacgc gcagactgtc gacggccccc
     3061 ccgaccgatg tcagcctggg ggacgagctc cacttagacg gcgaggacgt ggcgatggcg
     3121 catgccgacg cgctagacga tttcgatctg gacatgttgg gggacgggga ttccccgggt
     3181 ccgggattta ccccccacga ctccgccccc tacggcgctc tggatatggc cgacttcgag
     3241 tttgagcaga tgtttaccga tgcccttgga attgacgagt acggtgggta ggcggccgct
     3301 cataaagctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3361 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3421 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3481 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtccgg actgtacaat
     3541 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa
     3601 aacctctaca aatgtggtaa aatcgataag gatcttccta gagcatggct acgtagataa
     3661 gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc
     3721 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     3781 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcattaatg aatcggccaa
     3841 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3901 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3961 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4021 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4081 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4141 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4201 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4261 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4321 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4381 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4441 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4501 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4561 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4621 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4681 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4741 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4801 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4861 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4921 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4981 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5041 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5101 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5161 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5221 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5281 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5341 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5401 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5461 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5521 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5581 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5641 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5701 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5761 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     5821 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     5881 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     5941 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     6001 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     6061 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     6121 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     6181 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     6241 atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc
     6301 gatcggtgcg ggcctcttcg ctattacgcc agctgcgcgc tcgctcgctc actgaggccg
     6361 cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag
     6421 cgcgcagaga gggagtggcc aactccatca ctaggggttc cttgtagtta atgattaacc
     6481 cgccatgcta cttatctacg tagccatgct ctaggaagat cgtaggttaa ttaatctaga
     6541 ctgcagaggg ccctgcgtat gagtgcaagt gggttttagg accaggatga ggcggggtgg
     6601 gggtgcctac ctgacgaccg accccgaccc actggacaag cacccaaccc ccattcccca
     6661 aattgcgcat cccctatcag agagggggag gggaaacagg atgcggcgag gcgcgtgcgc
     6721 actgccagct tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg
     6781 ccgcctcagc actgaaggcg cgctgacgtc actcgccggt cccccgcaaa ctccccttcc
     6841 cggccacctt ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg
     6901 cgagataggg gggcacgggc gcgaccatct gcgctgcggc gccggcgact cagcgctgcc
     6961 tcagtctgcg gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt cgagaggatc
     7021 t
//
