LOCUS       sequence-86306-e        7889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Flpo in Mammalian Cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7889)
  AUTHORS   Fenno LE, Mattis J, Ramakrishnan C, Hyun M, Lee SY, He M, Tucciarone
            J, Selimbeyoglu A, Berndt A, Grosenick L, Zalocusky KA, Bernstein H,
            Swanson H, Perry C, Diester I, Boyce FM, Bass CE, Neve R, Huang ZJ, 
            Deisseroth K
  TITLE     Targeting cells with single vectors using multiple-feature Boolean 
            logic.
  JOURNAL   Nat Methods. 2014 Jul;11(7):763-72. doi: 10.1038/nmeth.2996. Epub 
            2014 Jun 8.
  PUBMED    24908100
REFERENCE   2  (bases 1 to 7889)
  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.2996"; journalName: "Nat Methods"; date: "2014-07"; 
            volume: "11"; issue: "7"; pages: "763-72"
COMMENT     Sequence Label: pAAV-EF1a-mCherry-IRES-Flpo
FEATURES             Location/Qualifiers
     source          1..7889
                     /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"
     CDS             1432..2139
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1574..1592)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1842..1861
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2156..2708
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2300..2317)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2527..2546
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    4016..4604
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4069..4089)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4487..4498)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4636..5112
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4835..4854)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5152..5292
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5152..5281
                     /label=AAV2 ITR
     rep_origin      5367..5822
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5454..5473)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5664..5685
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5839..5858)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5958..5980
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6018..6036)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6104..6208
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6209..6277,6278..7069)
                     /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(6427..6446)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7240..7828
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7729..7748
                     /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 cgtgagcaag
     1441 ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag
     1501 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     1561 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
     1621 atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
     1681 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     1741 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
     1801 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
     1861 accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc
     1921 gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc
     1981 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg
     2041 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
     2101 cgccactcca ccggcggcat ggacgagctg tacaagtaag gcgcgccccc ccctaacgtt
     2161 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     2221 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     2281 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     2341 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     2401 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     2461 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     2521 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     2581 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg
     2641 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     2701 tgataatatg gccacaacca tgagccagtt cgacatcctg tgcaagaccc cccccaaggt
     2761 gctggtgcgg cagttcgtgg agagattcga gaggcccagc ggcgagaaga tcgccagctg
     2821 tgccgccgag ctgacctacc tgtgctggat gatcacccac aacggcaccg ccatcaagag
     2881 ggccaccttc atgagctaca acaccatcat cagcaacagc ctgagcttcg acatcgtgaa
     2941 caagagcctg cagttcaagt acaagaccca gaaggccacc atcctggagg ccagcctgaa
     3001 gaagctgatc cccgcctggg agttcaccat catcccttac aacggccaga agcaccagag
     3061 cgacatcacc gacatcgtgt ccagcctgca gctgcagttc gagagcagcg aggaggccga
     3121 caagggcaac agccacagca agaagatgct gaaggccctg ctgtccgagg gcgagagcat
     3181 ctgggagatc accgagaaga tcctgaacag cttcgagtac accagcaggt tcaccaagac
     3241 caagaccctg taccagttcc tgttcctggc cacattcatc aactgcggca ggttcagcga
     3301 catcaagaac gtggacccca agagcttcaa gctggtgcag aacaagtacc tgggcgtgat
     3361 cattcagtgc ctggtgaccg agaccaagac aagcgtgtcc aggcacatct actttttcag
     3421 cgccagaggc aggatcgacc ccctggtgta cctggacgag ttcctgagga acagcgagcc
     3481 cgtgctgaag agagtgaaca ggaccggcaa cagcagcagc aacaagcagg agtaccagct
     3541 gctgaaggac aacctggtgc gcagctacaa caaggccctg aagaagaacg ccccctaccc
     3601 catcttcgct atcaagaacg gccctaagag ccacatcggc aggcacctga tgaccagctt
     3661 tctgagcatg aagggcctga ccgagctgac aaacgtggtg ggcaactgga gcgacaagag
     3721 ggcctccgcc gtggccagga ccacctacac ccaccagatc accgccatcc ccgaccacta
     3781 cttcgccctg gtgtccaggt actacgccta cgaccccatc agcaaggaga tgatcgccct
     3841 gaaggacgag accaacccca tcgaggagtg gcagcacatc gagcagctga agggcagcgc
     3901 cgagggcagc atcagatacc ccgcctggaa cggcatcatc agccaggagg tgctggacta
     3961 cctgagcagc tacatcaaca ggcggatctg agaattcgat atcaagctta tcgataatca
     4021 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     4081 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     4141 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     4201 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     4261 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     4321 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     4381 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     4441 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     4501 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     4561 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct
     4621 gctcgagaga tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
     4681 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
     4741 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
     4801 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
     4861 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
     4921 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
     4981 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag
     5041 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc
     5101 ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc
     5161 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     5221 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     5281 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     5341 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     5401 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     5461 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5521 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5581 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5641 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5701 tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     5761 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     5821 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     5881 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     5941 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     6001 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     6061 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     6121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     6181 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     6241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     6301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     6361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     6421 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     6481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     6541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     6601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     6661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     6721 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     6781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     6841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     6901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     6961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     7021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     7081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     7141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     7201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     7261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     7321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     7381 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     7441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     7501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     7561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     7621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     7681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     7741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     7801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     7861 ggccttttgc tggccttttg ctcacatgt
//
