LOCUS       sequence-177073-        7117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses iTango bArrestin2-TevC component and TdTomato 
            bicistronically.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7117)
  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 7117)
  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-bArrestin2-TevC-P2A-TdTomato
FEATURES             Location/Qualifiers
     source          1..7117
                     /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"
     CDS             2614..2670
                     /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             2677..4107
                     /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"
     primer_bind     complement(4124..4143)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4125..4143)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    4148..4282
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4172..4191)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4226..4245
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4395..4415)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4395..4413)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4434..4450)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4434..4450)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4447..4469)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4458..4474
                     /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(4482..4488,4489..4506,4507..4512))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4527..4548
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4665..4682)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4835..5423)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4915..4934)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5594..6385,6386..6454))
                     /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     6217..6236
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6455..6559)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6585..7040)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6722..6743)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6934..6953
                     /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 tatgggtgaa
     1021 aaacccggga ccagggtctt caagaagtcg agccctaact gcaagctcac cgtgtacttg
     1081 ggcaagcgtg actttgtgga tcacttggac aaagtggatc ctgtcgatgg tgtggtgctt
     1141 gtggatcctg actacttgaa ggaccggaaa gtgtttgtga ccctcacctg tgccttccgc
     1201 tatggccgag aagacctgga tgtactgggc ctgtctttcc gcaaagatct gttcatcgcc
     1261 acctaccagg ccttcccccc catgcccaac ccacctcggc cccccacccg cctacaggac
     1321 cgactgctga agaagttggg ccagcacgcc cacccctttt ttttcacaat accccagaat
     1381 ttgccttgct ccgtcacact gcagccagga ccggaggaca cagggaaggc ctgtggagta
     1441 gactttgaga ttcgagcctt ctgtgccaaa tctatagaag aaaaaagcca caaaaggaac
     1501 tccgtgcggc ttatcatcag aaaggtacag tttgctcctg agacacccgg cccccagcca
     1561 tcagctgaaa ccacacgcca cttcctcatg tctgaccgga ggtccctgca cctagaggct
     1621 tccctggaca aagagctgta ctaccatggg gaacccctca atgtcaacgt ccacgtcacc
     1681 aacaattctg ccaaaaccgt caagaagatc agagtgtctg tgagacagta tgccgacatt
     1741 tgcctcttca gcaccgcgca gtacaagtgt cctgtggctc agcttgaaca agatgaccag
     1801 gtgtctccca gttccacatt ctgcaaggtg tacaccataa ccccgctgct cagtgacaac
     1861 cgagagaagc gtggccttgc ccttgatggg caactcaagc acgaagacac caacctggct
     1921 tccagcacca ttgtgaagga gggagccaac aaggaggtgc tgggaatcct agtatcctac
     1981 agggtcaagg tgaagctggt ggtgtctcga ggcggggatg tctccgtgga gctacctttc
     2041 gtcctaatgc accccaagcc ccacgaccac atcacccttc cccgacccca gtcagccccc
     2101 cgggaaatag acatccctgt ggataccaac ctcattgaat tcgataccaa ctatgccaca
     2161 gacgacgaca tcgtgtttga ggactttgcg aggcttcggc tgaaggggat gaaggatgac
     2221 gactgtgatg accagttctg cctgagctca ctcgaaggtg gcggtggctc tggaggtggt
     2281 gggtccggag gaggcggcat cgagatgtcc agcatggtga gcgatactag ctgtaccttc
     2341 ccatcatctg acggaatctt ctggaagcac tggattcaga ctaaggacgg ccagtgtggc
     2401 agcccactgg tgagcacacg agacggattc atcgtgggga ttcacagcgc ctccaacttt
     2461 acaaacacca ataactattt cacctcagtg ccaaagaact ttatggagct gctgaccaac
     2521 caggaggccc agcagtgggt gagcgggtgg cgcctgaacg ccgattccgt gctgtggggc
     2581 gggcacaagg tgtttatggc atgcggaagc ggagctacta acttcagcct gctgaagcag
     2641 gctggagacg tggaggagaa ccctggacct gaattcatgg tgagcaaggg cgaggaggtc
     2701 atcaaagagt tcatgcgctt caaggtgcgc atggagggct ccatgaacgg ccacgagttc
     2761 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     2821 gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttcatgtac
     2881 ggctccaagg cgtacgtgaa gcaccccgcc gacatccccg attacaagaa gctgtccttc
     2941 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggtct ggtgaccgtg
     3001 acccaggact cctccctgca ggacggcacg ctgatctaca aggtgaagat gcgcggcacc
     3061 aacttccccc ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctccacc
     3121 gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccaccaggc cctgaagctg
     3181 aaggacggcg gccactacct ggtggagttc aagaccatct acatggccaa gaagcccgtg
     3241 caactgcccg gctactacta cgtggacacc aagctggaca tcacctccca caacgaggac
     3301 tacaccatcg tggaacagta cgagcgctcc gagggccgcc accacctgtt cctggggcat
     3361 ggcaccggca gcaccggcag cggcagctcc ggcaccgcct cctccgagga caacaacatg
     3421 gccgtcatca aagagttcat gcgcttcaag gtgcgcatgg agggctccat gaacggccac
     3481 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     3541 ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc
     3601 atgtacggct ccaaggcgta cgtgaagcac cccgccgaca tccccgatta caagaagctg
     3661 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg
     3721 accgtgaccc aggactcctc cctgcaggac ggcacgctga tctacaaggt gaagatgcgc
     3781 ggcaccaact tcccccccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     3841 tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg
     3901 aagctgaagg acggcggccg ctacctggtg gagttcaaga ccatctacat ggccaagaag
     3961 cccgtgcaac tgcccggcta ctactacgtg gacaccaagc tggacatcac ctcccacaac
     4021 gaggactaca ccatcgtgga acagtacgag cgctccgagg gccgccacca cctgttcctg
     4081 tacggcatgg acgagctgta caagtagtct agatagctag cctccctata gtgagtcgta
     4141 ttacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     4201 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     4261 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     4321 gggggaggtg tgggaggttt tttaattcgc gtccgcggcg ccaatgcatt gggcccggta
     4381 cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     4441 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccgggagc
     4501 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4561 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4621 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4681 ctgcgctcgg tcgttcggct gcgggagcgg tatcagctca ctcaaaggcg gtaatacggt
     4741 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4801 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4861 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4921 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4981 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5041 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5101 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5161 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5221 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5281 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5341 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5401 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5461 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5521 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5581 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5641 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5701 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5761 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5821 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5881 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5941 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6001 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6061 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6121 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6181 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6241 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6301 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6361 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6421 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6481 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6541 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
     6601 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     6661 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     6721 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     6781 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     6841 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     6901 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     6961 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     7021 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
     7081 atcggtgcgg gcctcttcgc tattacgcca gtcgacc
//
