LOCUS       sequence-469613-        7929 bp    DNA     circular SYN 22-APR-2025
DEFINITION  Flp-dependent expression of mNaChBacMut-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 Apr 22, 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-mNaChBacMut-T2A-tdTomato
FEATURES             Location/Qualifiers
     source          1..7929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(84..103)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      897..1485
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1386..1405
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1547..1676
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        1777..2955
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2008..2946
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2903..2923
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    3046..3079
                     /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(3086..4516)
                     /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      4513..4522
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4532..4585)
                     /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(5542..5575)
                     /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    5602..6190
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5655..5675)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6073..6084)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6222..6698
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(6421..6440)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   6738..6878
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6738..6867
                     /label=AAV2 ITR
     rep_origin      6953..7408
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7040..7059)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7250..7271
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7425..7444)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7544..7566
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7604..7622)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7690..7794
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7795..7863,7864..7929,1..726)
                     /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"
ORIGIN
        1 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
       61 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
      121 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
      181 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
      241 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
      301 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
      361 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
      421 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
      481 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
      541 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
      601 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
      661 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
      721 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
      781 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
      841 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
      901 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
      961 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1021 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1081 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1141 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1201 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1261 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1321 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1381 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1441 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1501 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg
     1561 cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc
     1621 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg
     1681 ccgcacgcgt aagctttgca aagatggata aagttttaaa cagagaggaa tctttgcagc
     1741 taatggacct tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca
     1801 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt
     1861 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
     1921 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
     1981 cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc
     2041 ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg
     2101 attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa
     2161 ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg
     2221 cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa
     2281 aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc
     2341 caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg
     2401 tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg
     2461 gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc
     2521 ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg
     2581 cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg
     2641 ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac
     2701 tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg
     2761 tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg
     2821 gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt
     2881 gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca
     2941 tttcaggtgt cgtgaggtac cgaagttcct attctatcag aagtatagga acttcgcaga
     3001 atggtagctg gattgtagct gctattagca atatgaaacc tcttagaagt tcctattctc
     3061 tagaaagtat aggaacttcg aattcttact tgtacagctc gtccatgccg tacaggaaca
     3121 ggtggtggcg gccctcggag cgctcgtact gttccacgat ggtgtagtcc tcgttgtggg
     3181 aggtgatgtc cagcttggtg tccacgtagt agtagccggg cagttgcacg ggcttcttgg
     3241 ccatgtagat ggtcttgaac tccaccaggt agtggccgcc gtccttcagc ttcagggcct
     3301 ggtggatctc gcccttcagc acgccgtcgc gggggtacag gcgctcggtg gaggcctccc
     3361 agcccatggt cttcttctgc attacggggc cgtcgggggg gaagttggtg ccgcgcatct
     3421 tcaccttgta gatcagcgtg ccgtcctgca gggaggagtc ctgggtcacg gtcaccagac
     3481 cgccgtcctc gaagttcatc acgcgctccc acttgaagcc ctcggggaag gacagcttct
     3541 tgtaatcggg gatgtcggcg gggtgcttca cgtacgcctt ggagccgtac atgaactggg
     3601 gggacaggat gtcccaggcg aagggcaggg ggccgccctt ggtcaccttc agcttggcgg
     3661 tctgggtgcc ctcgtagggg cggccctcgc cctcgccctc gatctcgaac tcgtggccgt
     3721 tcatggagcc ctccatgcgc accttgaagc gcatgaactc tttgatgacg gccatgttgt
     3781 tgtcctcgga ggaggcggtg ccggagctgc cgctgccggt gctgccggtg ccatgcccca
     3841 ggaacaggtg gtggcggccc tcggagcgct cgtactgttc cacgatggtg tagtcctcgt
     3901 tgtgggaggt gatgtccagc ttggtgtcca cgtagtagta gccgggcagt tgcacgggct
     3961 tcttggccat gtagatggtc ttgaactcca ccaggtagtg gccgccgtcc ttcagcttca
     4021 gggcctggtg gatctcgccc ttcagcacgc cgtcgcgggg gtacaggcgc tcggtggagg
     4081 cctcccagcc catggtcttc ttctgcatta cggggccgtc gggggggaag ttggtgccgc
     4141 gcatcttcac cttgtagatc agcgtgccgt cctgcaggga ggagtcctgg gtcacggtca
     4201 ccagaccgcc gtcctcgaag ttcatcacgc gctcccactt gaagccctcg gggaaggaca
     4261 gcttcttgta atcggggatg tcggcggggt gcttcacgta cgccttggag ccgtacatga
     4321 actgggggga caggatgtcc caggcgaagg gcagggggcc gcccttggtc accttcagct
     4381 tggcggtctg ggtgccctcg taggggcggc cctcgccctc gccctcgatc tcgaactcgt
     4441 ggccgttcat ggagccctcc atgcgcacct tgaagcgcat gaactctttg atgacctcct
     4501 cgcccttgct caccatggtg gcgggatccg ctgggccagg attctcctcg acgtcaccgc
     4561 atgttagcag acttcctctg ccctctccac tgccgatatc cttaagttgt ttgagcaagc
     4621 tcttcagttc ggctacatcc tttcgcaatg ctgaaatctc ctgtttaagt ccgtcggcct
     4681 ccccatcttc ctcattgtcc gtaagctccg ccttttccac gttgttcaca atgactccga
     4741 tgaaaagatt gaaaatgata aaagtaccaa tcaggacaaa cgagacaaag tagagccatg
     4801 accagggcac ctctgcaaaa atgggccgca ttacaccgct ggcccagctt ttcaaggtca
     4861 ccacttggaa cagcgtcagc aacgaaagct ggaggttccc aaagtattca ggcgatacgt
     4921 gctggaagag cattgtcccg atcactgcaa agatatagaa gaagatggac atcaagatca
     4981 agatattgcc caacgccgga atcgtcatca ccagcgcatc tacgagtctg cgcagggagg
     5041 ggacgacgga gatcgccctg agtacccgca gcactctaag gattcgcaat actgtcacga
     5101 actgggctcc cgcgaagata tgcccagccg cgacaatcag aaaatcaaac caattccacg
     5161 agctgcggaa gaaggccgac tttgggttcg acgccaaaaa cctcatcgcg atctcgatgg
     5221 tgaagatcca aagcaggacg aggtcaatcc gataaaagag ccacttgtgg tccgcgtaga
     5281 tcctcggata tgtttcgatt cccacgatca gtgcattaaa gaggatcagg gcaatgacgg
     5341 tgaaagtaaa cgctctatgg ttgacaattt tctgcatctt tgacgtgaat gagttctgtt
     5401 tctggcgtgc ttccattttc atggtggcgg ctagctacgt aggcgcgcct taattaagaa
     5461 gttcctatac ttctgataga ataggaactt cttgccttaa cccagaaatt atcactgtta
     5521 ttctttagaa tggtgcaaag agaagttcct atactttcta gagaatagga acttccagaa
     5581 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     5641 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     5701 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     5761 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     5821 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     5881 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     5941 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     6001 tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc
     6061 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     6121 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     6181 gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat ccctgtgacc
     6241 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     6301 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     6361 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     6421 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     6481 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     6541 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     6601 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     6661 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag gtaaccacgt
     6721 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     6781 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     6841 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     6901 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     6961 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     7021 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     7081 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     7141 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     7201 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     7261 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     7321 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     7381 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     7441 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     7501 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     7561 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     7621 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     7681 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     7741 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     7801 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     7861 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     7921 ggttacatc
//
