LOCUS       sequence-177108-        7466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses iTango bArrestin2-TevC component and TdTomato 
            bicistronically for AAV production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7466)
  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 7466)
  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-bArrestin2-TevC-P2A-TdTomato-WPRE-bGHpA
FEATURES             Location/Qualifiers
     source          1..7466
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1609..1665
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1672..3102
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGRYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     misc_feature    3129..3717
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3182..3202)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3600..3611)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3758..3965
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4501..5089)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4581..4600)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5260..6051,6052..6120))
                     /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     5883..5902
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6121..6225)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6251..6706)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6388..6409)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6600..6619
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6778..6907
                     /label=AAV2 ITR
     promoter        7007..7454
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 aagcttatgg gtgaaaaacc cgggaccagg gtcttcaaga agtcgagccc taactgcaag
       61 ctcaccgtgt acttgggcaa gcgtgacttt gtggatcact tggacaaagt ggatcctgtc
      121 gatggtgtgg tgcttgtgga tcctgactac ttgaaggacc ggaaagtgtt tgtgaccctc
      181 acctgtgcct tccgctatgg ccgagaagac ctggatgtac tgggcctgtc tttccgcaaa
      241 gatctgttca tcgccaccta ccaggccttc ccccccatgc ccaacccacc tcggcccccc
      301 acccgcctac aggaccgact gctgaagaag ttgggccagc acgcccaccc cttttttttc
      361 acaatacccc agaatttgcc ttgctccgtc acactgcagc caggaccgga ggacacaggg
      421 aaggcctgtg gagtagactt tgagattcga gccttctgtg ccaaatctat agaagaaaaa
      481 agccacaaaa ggaactccgt gcggcttatc atcagaaagg tacagtttgc tcctgagaca
      541 cccggccccc agccatcagc tgaaaccaca cgccacttcc tcatgtctga ccggaggtcc
      601 ctgcacctag aggcttccct ggacaaagag ctgtactacc atggggaacc cctcaatgtc
      661 aacgtccacg tcaccaacaa ttctgccaaa accgtcaaga agatcagagt gtctgtgaga
      721 cagtatgccg acatttgcct cttcagcacc gcgcagtaca agtgtcctgt ggctcagctt
      781 gaacaagatg accaggtgtc tcccagttcc acattctgca aggtgtacac cataaccccg
      841 ctgctcagtg acaaccgaga gaagcgtggc cttgcccttg atgggcaact caagcacgaa
      901 gacaccaacc tggcttccag caccattgtg aaggagggag ccaacaagga ggtgctggga
      961 atcctagtat cctacagggt caaggtgaag ctggtggtgt ctcgaggcgg ggatgtctcc
     1021 gtggagctac ctttcgtcct aatgcacccc aagccccacg accacatcac ccttccccga
     1081 ccccagtcag ccccccggga aatagacatc cctgtggata ccaacctcat tgaattcgat
     1141 accaactatg ccacagacga cgacatcgtg tttgaggact ttgcgaggct tcggctgaag
     1201 gggatgaagg atgacgactg tgatgaccag ttctgcctga gctcactcga aggtggcggt
     1261 ggctctggag gtggtgggtc cggaggaggc ggcatcgaga tgtccagcat ggtgagcgat
     1321 actagctgta ccttcccatc atctgacgga atcttctgga agcactggat tcagactaag
     1381 gacggccagt gtggcagccc actggtgagc acacgagacg gattcatcgt ggggattcac
     1441 agcgcctcca actttacaaa caccaataac tatttcacct cagtgccaaa gaactttatg
     1501 gagctgctga ccaaccagga ggcccagcag tgggtgagcg ggtggcgcct gaacgccgat
     1561 tccgtgctgt ggggcgggca caaggtgttt atggcatgcg gaagcggagc tactaacttc
     1621 agcctgctga agcaggctgg agacgtggag gagaaccctg gacctgaatt catggtgagc
     1681 aagggcgagg aggtcatcaa agagttcatg cgcttcaagg tgcgcatgga gggctccatg
     1741 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     1801 accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc
     1861 ccccagttca tgtacggctc caaggcgtac gtgaagcacc ccgccgacat ccccgattac
     1921 aagaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     1981 ggtctggtga ccgtgaccca ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg
     2041 aagatgcgcg gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gaccatgggc
     2101 tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg cgagatccac
     2161 caggccctga agctgaagga cggcggccac tacctggtgg agttcaagac catctacatg
     2221 gccaagaagc ccgtgcaact gcccggctac tactacgtgg acaccaagct ggacatcacc
     2281 tcccacaacg aggactacac catcgtggaa cagtacgagc gctccgaggg ccgccaccac
     2341 ctgttcctgg ggcatggcac cggcagcacc ggcagcggca gctccggcac cgcctcctcc
     2401 gaggacaaca acatggccgt catcaaagag ttcatgcgct tcaaggtgcg catggagggc
     2461 tccatgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     2521 acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc
     2581 ctgtcccccc agttcatgta cggctccaag gcgtacgtga agcaccccgc cgacatcccc
     2641 gattacaaga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     2701 gacggcggtc tggtgaccgt gacccaggac tcctccctgc aggacggcac gctgatctac
     2761 aaggtgaaga tgcgcggcac caacttcccc cccgacggcc ccgtaatgca gaagaagacc
     2821 atgggctggg aggcctccac cgagcgcctg tacccccgcg acggcgtgct gaagggcgag
     2881 atccaccagg ccctgaagct gaaggacggc ggccgctacc tggtggagtt caagaccatc
     2941 tacatggcca agaagcccgt gcaactgccc ggctactact acgtggacac caagctggac
     3001 atcacctccc acaacgagga ctacaccatc gtggaacagt acgagcgctc cgagggccgc
     3061 caccacctgt tcctgtacgg catggacgag ctgtacaagt agtctagata gctagccagc
     3121 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     3181 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     3241 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     3301 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     3361 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     3421 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     3481 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     3541 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     3601 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     3661 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     3721 gataccgtcg acccgggcgg ccgctcataa agctcgactg tgccttctag ttgccagcca
     3781 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3841 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3901 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3961 ggggatgcgg tccggactgt acaattgcat tcattttatg tttcaggttc agggggagat
     4021 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct
     4081 tcctagagca tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac
     4141 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgccc
     4201 gggctntncm ckgnggccgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     4261 gcagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4321 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4381 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4441 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4501 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4561 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4621 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4681 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4741 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4801 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4861 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4921 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4981 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5041 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5101 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5161 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5221 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5281 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5341 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5401 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5461 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5521 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5581 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5641 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5701 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5761 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5821 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5881 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5941 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6001 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6061 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6121 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6181 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6241 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     6301 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     6361 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     6421 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     6481 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     6541 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     6601 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     6661 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
     6721 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     6781 cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt
     6841 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
     6901 ggttccttgt agttaatgat taacccgcca tgctacttat ctacgtagcc atgctctagg
     6961 aagatcgtag gttaattaat ctagactgca gagggccctg cgtatgagtg caagtgggtt
     7021 ttaggaccag gatgaggcgg ggtgggggtg cctacctgac gaccgacccc gacccactgg
     7081 acaagcaccc aacccccatt ccccaaattg cgcatcccct atcagagagg gggaggggaa
     7141 acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca gtgccttcgc
     7201 ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg acgtcactcg
     7261 ccggtccccc gcaaactccc cttcccggcc accttggtcg cgtccgcgcc gccgccggcc
     7321 cagccggacc gcaccacgcg aggcgcgaga taggggggca cgggcgcgac catctgcgct
     7381 gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg cagcggagga gtcgtgtcgt
     7441 gcctgagagc gcagtcgaga ggatct
//
