LOCUS       sequence-354763-        7929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flp-dependent expression of mNaChBac-T2A-tdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7929)
  AUTHORS   Xue M, Atallah BV, Scanziani M
  TITLE     Equalizing excitation-inhibition ratios across visual cortical 
            neurons.
  JOURNAL   Nature. 2014 Jul 31;511(7511):596-600. doi: 10.1038/nature13321. 
            Epub 2014 Jun 22.
  PUBMED    25043046
REFERENCE   2  (bases 1 to 7929)
  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/nature13321"; journalName: "Nature"; date: "2014-07-31- 
            31"; volume: "511"; issue: "7511"; pages: "596-600"
COMMENT     Sequence Label: pAAV-EF1-alpha-F-FLEX-mNaChBac-T2A-tdTomato
FEATURES             Location/Qualifiers
     source          1..7929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1500..1533
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             complement(1540..2970)
                     /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"
     regulatory      2967..2976
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2986..3039)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     protein_bind    complement(3996..4029)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    4056..4644
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4109..4129)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4527..4538)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4676..5152
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4875..4894)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5192..5332
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5192..5321
                     /label=AAV2 ITR
     rep_origin      5407..5862
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5494..5513)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5704..5725
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5879..5898)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5998..6020
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6058..6076)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6144..6248
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6249..6317,6318..7109)
                     /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(6467..6486)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7280..7868
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7769..7788
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccgaagt tcctattcta tcagaagtat
     1441 aggaacttcg cagaatggta gctggattgt agctgctatt agcaatatga aacctcttag
     1501 aagttcctat tctctagaaa gtataggaac ttcgaattct tacttgtaca gctcgtccat
     1561 gccgtacagg aacaggtggt ggcggccctc ggagcgctcg tactgttcca cgatggtgta
     1621 gtcctcgttg tgggaggtga tgtccagctt ggtgtccacg tagtagtagc cgggcagttg
     1681 cacgggcttc ttggccatgt agatggtctt gaactccacc aggtagtggc cgccgtcctt
     1741 cagcttcagg gcctggtgga tctcgccctt cagcacgccg tcgcgggggt acaggcgctc
     1801 ggtggaggcc tcccagccca tggtcttctt ctgcattacg gggccgtcgg gggggaagtt
     1861 ggtgccgcgc atcttcacct tgtagatcag cgtgccgtcc tgcagggagg agtcctgggt
     1921 cacggtcacc agaccgccgt cctcgaagtt catcacgcgc tcccacttga agccctcggg
     1981 gaaggacagc ttcttgtaat cggggatgtc ggcggggtgc ttcacgtacg ccttggagcc
     2041 gtacatgaac tggggggaca ggatgtccca ggcgaagggc agggggccgc ccttggtcac
     2101 cttcagcttg gcggtctggg tgccctcgta ggggcggccc tcgccctcgc cctcgatctc
     2161 gaactcgtgg ccgttcatgg agccctccat gcgcaccttg aagcgcatga actctttgat
     2221 gacggccatg ttgttgtcct cggaggaggc ggtgccggag ctgccgctgc cggtgctgcc
     2281 ggtgccatgc cccaggaaca ggtggtggcg gccctcggag cgctcgtact gttccacgat
     2341 ggtgtagtcc tcgttgtggg aggtgatgtc cagcttggtg tccacgtagt agtagccggg
     2401 cagttgcacg ggcttcttgg ccatgtagat ggtcttgaac tccaccaggt agtggccgcc
     2461 gtccttcagc ttcagggcct ggtggatctc gcccttcagc acgccgtcgc gggggtacag
     2521 gcgctcggtg gaggcctccc agcccatggt cttcttctgc attacggggc cgtcgggggg
     2581 gaagttggtg ccgcgcatct tcaccttgta gatcagcgtg ccgtcctgca gggaggagtc
     2641 ctgggtcacg gtcaccagac cgccgtcctc gaagttcatc acgcgctccc acttgaagcc
     2701 ctcggggaag gacagcttct tgtaatcggg gatgtcggcg gggtgcttca cgtacgcctt
     2761 ggagccgtac atgaactggg gggacaggat gtcccaggcg aagggcaggg ggccgccctt
     2821 ggtcaccttc agcttggcgg tctgggtgcc ctcgtagggg cggccctcgc cctcgccctc
     2881 gatctcgaac tcgtggccgt tcatggagcc ctccatgcgc accttgaagc gcatgaactc
     2941 tttgatgacc tcctcgccct tgctcaccat ggtggcggga tccgctgggc caggattctc
     3001 ctcgacgtca ccgcatgtta gcagacttcc tctgccctct ccactgccga tatccttaag
     3061 ttgtttgagc aagctcttca gttcggctac atcctttcgc aatgctgaaa tctcctgttt
     3121 aagtccgtcg gcctccccat cttcctcatt gtccgtaagc tccgcctttt ccacgttgtt
     3181 cacaatgact ccgatgaaaa gattgaaaat gataaaagta ccaatcagga caaacgagac
     3241 aaagtagagc catgaccagg gcacctctgc aaaaatgggc cgcattacac cgctggccca
     3301 gctctccaag gtcaccactt ggaacagcgt cagcaacgaa agctggaggt tcccaaagta
     3361 ttcaggcgat acgtgctgga agagcattgt cccgatcact gcaaagatat agaagaagat
     3421 ggacatcaag atcaagatat tgcccaacgc cggaatcgtc atcaccagcg catctacgag
     3481 tctgcgcagg gaggggacga cggagatcgc cctgagtacc cgcagcactc taaggattcg
     3541 caatactgtc acgaactggg ctcccgcgaa gatatgccca gccgcgacaa tcagaaaatc
     3601 aaaccaattc cacgagctgc ggaagaaggc cgactttggg ttcgacgcca aaaacctcat
     3661 cgcgatctcg atggtgaaga tccaaagcag gacgaggtca atccgataaa agagccactt
     3721 gtggtccgcg tagatcctcg gatatgtttc gattcccacg atcagtgcat taaagaggat
     3781 cagggcaatg acggtgaaag taaacgctct atggttgaca attttctgca tctttgacgt
     3841 gaatgagttc tgtttctggc gtgcttccat tttcatggtg gcggctagct acgtaggcgc
     3901 gccttaatta agaagttcct atacttctga tagaatagga acttcttgcc ttaacccaga
     3961 aattatcact gttattcttt agaatggtgc aaagagaagt tcctatactt tctagagaat
     4021 aggaacttcc agaattcgat atcaagctta tcgataatca acctctggat tacaaaattt
     4081 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     4141 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     4201 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     4261 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     4321 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     4381 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     4441 tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc
     4501 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     4561 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     4621 tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg
     4681 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
     4741 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
     4801 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
     4861 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
     4921 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg
     4981 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt
     5041 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc
     5101 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt
     5161 gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     5221 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     5281 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     5341 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     5401 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     5461 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     5521 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5581 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     5641 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5701 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     5761 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     5821 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     5881 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     5941 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     6001 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     6061 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     6121 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     6181 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     6241 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     6301 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     6361 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     6421 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     6481 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     6541 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     6601 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6661 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6721 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6781 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     6841 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     6901 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     6961 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     7021 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     7081 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     7141 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     7201 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     7261 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     7321 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     7381 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     7441 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     7501 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     7561 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     7621 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     7681 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     7741 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     7801 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     7861 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     7921 ctcacatgt
//
