LOCUS       sequence-116188-        6775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ""Expresses the fluorescent protein tdTomato"".
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6775)
  AUTHORS   Wertz A, Trenholm S, Yonehara K, Hillier D, Raics Z, Leinweber M, 
            Szalay G, Ghanem A, Keller G, Rozsa B, Conzelmann KK, Roska B
  TITLE     PRESYNAPTIC NETWORKS. Single-cell-initiated monosynaptic tracing 
            reveals layer-specific cortical network modules.
  JOURNAL   Science. 2015 Jul 3;349(6243):70-4. doi: 10.1126/science.aab1687.
  PUBMED    26138975
REFERENCE   2  (bases 1 to 6775)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2015-07-3- 3"; volume: "349"; issue: 
            "6243"; pages: "70-4"
COMMENT     Sequence Label: pAAV-EF1a-tdTomato-WPRE-pGHpA
FEATURES             Location/Qualifiers
     source          1..6775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1438..1447
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1444..2874
                     /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
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     misc_feature    2902..3490
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2955..2975)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3373..3384)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3522..3998
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3721..3740)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4038..4178
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4038..4167
                     /label=AAV2 ITR
     rep_origin      4253..4708
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4340..4359)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4550..4571
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4725..4744)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4844..4866
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4904..4922)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4990..5094
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5095..5163,5164..5955)
                     /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(5313..5332)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6126..6714
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6615..6634
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagcc
     1441 accatggtga gcaagggcga ggaggtcatc aaagagttca tgcgcttcaa ggtgcgcatg
     1501 gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     1561 gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg
     1621 gacatcctgt ccccccagtt catgtacggc tccaaggcgt acgtgaagca ccccgccgac
     1681 atccccgatt acaagaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     1741 ttcgaggacg gcggtctggt gaccgtgacc caggactcct ccctgcagga cggcacgctg
     1801 atctacaagg tgaagatgcg cggcaccaac ttcccccccg acggccccgt aatgcagaag
     1861 aagaccatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag
     1921 ggcgagatcc accaggccct gaagctgaag gacggcggcc actacctggt ggagttcaag
     1981 accatctaca tggccaagaa gcccgtgcaa ctgcccggct actactacgt ggacaccaag
     2041 ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga gcgctccgag
     2101 ggccgccacc acctgttcct ggggcatggc accggcagca ccggcagcgg cagctccggc
     2161 accgcctcct ccgaggacaa caacatggcc gtcatcaaag agttcatgcg cttcaaggtg
     2221 cgcatggagg gctccatgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     2281 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc
     2341 gcctgggaca tcctgtcccc ccagttcatg tacggctcca aggcgtacgt gaagcacccc
     2401 gccgacatcc ccgattacaa gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     2461 atgaacttcg aggacggcgg tctggtgacc gtgacccagg actcctccct gcaggacggc
     2521 acgctgatct acaaggtgaa gatgcgcggc accaacttcc cccccgacgg ccccgtaatg
     2581 cagaagaaga ccatgggctg ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg
     2641 ctgaagggcg agatccacca ggccctgaag ctgaaggacg gcggccacta cctggtggag
     2701 ttcaagacca tctacatggc caagaagccc gtgcaactgc ccggctacta ctacgtggac
     2761 accaagctgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc
     2821 tccgagggcc gccaccacct gttcctgtac ggcatggacg agctgtacaa gtagtaagaa
     2881 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     2941 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3001 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     3061 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3121 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3181 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3241 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     3301 tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc
     3361 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3421 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3481 gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat ccctgtgacc
     3541 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     3601 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     3661 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     3721 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     3781 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     3841 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     3901 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     3961 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag gtaaccacgt
     4021 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     4081 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     4141 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     4201 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     4261 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4321 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4381 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4441 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     4501 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4561 ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
     4621 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4681 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     4741 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4801 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4861 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     4921 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4981 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5041 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5101 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5161 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5221 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5281 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5341 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5401 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5461 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5521 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5581 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5641 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5701 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5761 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5821 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5881 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5941 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6001 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6061 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6121 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6181 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6241 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     6301 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6361 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6421 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6481 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6541 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6601 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6661 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6721 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
