LOCUS       sequence-182717-        7879 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 7879)
  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 7879)
  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..7879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1045..1597
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1207..1226)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     1436..1453
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(1614..2321)
                     /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(1892..1911)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2161..2179
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     promoter        complement(2334..3512)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2343..3281
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(2366..2386)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     repeat_region   3613..3742
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3804..4392)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3884..3903)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4563..5354,5355..5423))
                     /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     5186..5205
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5424..5528)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5596..5614
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5652..5674)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5774..5793
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5810..6265)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5947..5968)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6159..6178
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6340..6480
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6351..6480
                     /label=AAV2 ITR
     polyA_signal    6520..6996
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     6778..6797
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    7028..7616
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             7134..7145
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     7543..7563
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 gggtgtaggt ggtcctggcc acggcggagg ccctcttgtc gctccagttg cccaccacgt
       61 ttgtcagctc ggtcaggccc ttcatgctca gaaagctggt catcaggtgc ctgccgatgt
      121 ggctcttagg gccgttcttg atagcgaaga tggggtaggg ggcgttcttc ttcagggcct
      181 tgttgtagct gcgcaccagg ttgtccttca gcagctggta ctcctgcttg ttgctgctgc
      241 tgttgccggt cctgttcact ctcttcagca cgggctcgct gttcctcagg aactcgtcca
      301 ggtacaccag ggggtcgatc ctgcctctgg cgctgaaaaa gtagatgtgc ctggacacgc
      361 ttgtcttggt ctcggtcacc aggcactgaa tgatcacgcc caggtacttg ttctgcacca
      421 gcttgaagct cttggggtcc acgttcttga tgtcgctgaa cctgccgcag ttgatgaatg
      481 tggccaggaa caggaactgg tacagggtct tggtcttggt gaacctgctg gtgtactcga
      541 agctgttcag gatcttctcg gtgatctccc agatgctctc gccctcggac agcagggcct
      601 tcagcatctt cttgctgtgg ctgttgccct tgtcggcctc ctcgctgctc tcgaactgca
      661 gctgcaggct ggacacgatg tcggtgatgt cgctctggtg cttctggccg ttgtaaggga
      721 tgatggtgaa ctcccaggcg gggatcagct tcttcaggct ggcctccagg atggtggcct
      781 tctgggtctt gtacttgaac tgcaggctct tgttcacgat gtcgaagctc aggctgttgc
      841 tgatgatggt gttgtagctc atgaaggtgg ccctcttgat ggcggtgccg ttgtgggtga
      901 tcatccagca caggtaggtc agctcggcgg cacagctggc gatcttctcg ccgctgggcc
      961 tctcgaatct ctccacgaac tgccgcacca gcaccttggg gggggtcttg cacaggatgt
     1021 cgaactggct catggttgtg gccatattat catcgtgttt ttcaaaggaa aaccacgtcc
     1081 ccgtggttcg gggggcctag acgttttttt aacctcgact aaacacatgt aaagcatgtg
     1141 caccgaggcc ccagatcaga tcccatacaa tggggtacct tctgggcatc cttcagcccc
     1201 ttgttgaata cgcttgagga gagccatttg actctttcca caactatcca actcacaacg
     1261 tggcactggg gttgtgccgc ctttgcaggt gtatcttata cacgtggctt ttggccgcag
     1321 aggcacctgt cgccaggtgg ggggttccgc tgcctgcaaa gggtcgctac agacgttgtt
     1381 tgtcttcaag aagcttccag aggaactgct tccttcacga cattcaacag accttgcatt
     1441 cctttggcga gaggggaaag acccctagga atgctcgtca agaagacagg gccaggtttc
     1501 cgggccctca cattgccaaa agacggcaat atggtggaaa ataacatata gacaaacgca
     1561 caccggcctt attccaagcg gcttcggcca gtaacgttag gggggggcgc gccttacttg
     1621 tacagctcgt ccatgccgcc ggtggagtgg cggccctcgg cgcgttcgta ctgttccacg
     1681 atggtgtagt cctcgttgtg ggaggtgatg tccaacttga tgttgacgtt gtaggcgccg
     1741 ggcagctgca cgggcttctt ggccttgtag gtggtcttga cctcagcgtc gtagtggccg
     1801 ccgtccttca gcttcagcct ctgcttgatc tcgcccttca gggcgccgtc ctcggggtac
     1861 atccgctcgg aggaggcctc ccagcccatg gtcttcttct gcattacggg gccgtcggag
     1921 gggaagttgg tgccgcgcag cttcaccttg tagatgaact cgccgtcctg cagggaggag
     1981 tcctgggtca cggtcaccac gccgccgtcc tcgaagttca tcacgcgctc ccacttgaag
     2041 ccctcgggga aggacagctt caagtagtcg gggatgtcgg cggggtgctt cacgtaggcc
     2101 ttggagccgt acatgaactg aggggacagg atgtcccagg cgaagggcag ggggccaccc
     2161 ttggtcacct tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc gccctcgccc
     2221 tcgatctcga actcgtggcc gttcacggag ccctccatgt gcaccttgaa gcgcatgaac
     2281 tccttgatga tggccatgtt atcctcctcg cccttgctca cggatccggt acctcacgac
     2341 acctgaaatg gaagaaaaaa actttgaacc actgtctgag gcttgagaat gaaccaagat
     2401 ccaaactcaa aaagggcaaa ttccaaggag aattacatca agtgccaagc tggcctaact
     2461 tcagtctcca cccactcagt gtggggaaac tccatcgcat aaaacccctc cccccaacct
     2521 aaagacgacg tactccaaaa gctcgagaac taatcgaggt gcctggacgg cgcccggtac
     2581 tccgtggagt cacatgaagc gacggctgag gacggaaagg cccttttcct ttgtgtgggt
     2641 gactcacccg cccgctctcc cgagcgccgc gtcctccatt ttgagctccc tgcagcaggg
     2701 ccgggaagcg gccatctttc cgctcacgca actggtgccg accgggccag ccttgccgcc
     2761 cagggcgggg cgatacacgg cggcgcgagg ccaggcacca gagcaggccg gccagcttga
     2821 gactaccccc gtccgattct cggtggccgc gctcgcaggc cccgcctcgc cgaacatgtg
     2881 cgctgggacg cacgggcccc gtcgccgccc gcggccccaa aaaccgaaat accagtgtgc
     2941 agatcttggc ccgcatttac aagactatct tgccagaaaa aaagcgtcgc agcaggtcat
     3001 caaaaatttt aaatggctag agacttatcg aaagcagcga gacaggcgcg aaggtgccac
     3061 cagattcgca cgcggcggcc ccagcgccca ggccaggcct caactcaagc acgaggcgaa
     3121 ggggctcctt aagcgcaagg cctcgaactc tcccacccac ttccaacccg aagctcggga
     3181 tcaagaatca cgtactgcag ccaggtggaa gtaattcaag gcacgcaagg gccataaccc
     3241 gtaaagaggc caggcccgcg ggaaccacac acggcactta cctgtgttct ggcggcaaac
     3301 ccgttgcgaa aaagaacgtt cacggcgact actgcactta tatacggttc tcccccaccc
     3361 tcgggaaaaa ggcggagcca gtacacgaca tcactttccc agtttacccc gcgccacctt
     3421 ctctaggcac cggttcaatt gccgacccct ccccccaact tctcggggac tgtgggcgat
     3481 gtgcgctctg cccactgacg ggcaccggag ccaattccca ctcctttcaa gacctagaag
     3541 gtccattagc tgcaaagatt cctctctgtt taaaacttta tccatctttg caaagcttac
     3601 gcgtgcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     3661 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggcggcctc agtgagcgag
     3721 cgagcgcgca gctgcctgca ggacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3781 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3841 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3901 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3961 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4021 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4081 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4141 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4201 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4261 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4321 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4381 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4441 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4501 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4561 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4621 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4681 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4741 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4801 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4861 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4921 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4981 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5041 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5101 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5161 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5221 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5281 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5341 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5401 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5461 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5521 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     5581 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     5641 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     5701 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     5761 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccataa aattgtaaac
     5821 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     5881 tagaccgaaa tcggcaaaat cccttataaa tcaaaagaat agcccgagat agagttgagt
     5941 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     6001 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccaa atcaagtttt
     6061 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     6121 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     6181 ggcgctaagg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     6241 cttaatgcgc cgctacaggg cgcgtactat ggttgctttg acgtatgcgg tgtgaaatac
     6301 cgcacagatg cgtaaggaga aaataccgca tcaggcgccc ctgcaggcag ctgcgcgctc
     6361 gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg
     6421 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
     6481 gcggccgctc ggtccgcacg tggttaccta caaaatcaga aggacaggga agggagcagt
     6541 ggttcacgcc tgtaatccca gcaatttggg aggccaaggt gggtagatca cctgagatta
     6601 ggagttggag accagcctgg ccaatatggt gaaaccccgt ctctaccaaa aaaacaaaaa
     6661 ttagctgagc ctggtcatgc atgcctggaa tcccaacaac tcgggaggct gaggcaggag
     6721 aatcgcttga acccaggagg cggagattgc agtgagccaa gattgtgcca ctgcactcca
     6781 gcttggttcc caatagaccc cgcaggccct acaggttgtc ttcccaactt gccccttgct
     6841 ccataccacc cccctccacc ccataatatt atagaaggac acctagtcag acaaaatgat
     6901 gcaacttaat tttattagga caaggctggt gggcactgga gtggcaactt ccagggccag
     6961 gagaggcact ggggaggggt cacagggatg ccacccgtag atctctcgag cagcgctcgg
     7021 tatcgatgcg gggaggcggc ccaaagggag atccgactcg tctgagggcg aaggcgaaga
     7081 cgcggaagag gccgcagagc cggcagcagg ccgcgggaag gaaggtccgc tggattgagg
     7141 gccgaaggga cgtagcagaa ggacgtcccg cgcagaatcc aggtggcaac ataggcgagc
     7201 agccaaggaa aggacgatga tttccccgac aacaccacgg aattgtcagt gcccaacagc
     7261 cgagcccctg tccagcagcg ggcaaggcag gcggcgatga gttccgccgt ggcaataggg
     7321 agggggaaag cgaaagtccc ggaaaggagc tgacaggtgg tggcaatgcc ccaaccagtg
     7381 ggggttgcgt cagcaaacac agtgcacacc acgccacgtt gcctgacaac gggccacaac
     7441 tcctcataaa gagacagcaa ccaggattta tacaaggagg agaaaatgaa agccatacgg
     7501 gaagcaatag catgatacaa aggcattaaa gcagcgtatc cacatagcgt aaaaggagca
     7561 acatagttaa gaataccagt caatctttca caaattttgt aatccagagg ttgattatcg
     7621 ataagcttga tatcgaattc tcagatccgc ctgttgatgt agctgctcag gtagtccagc
     7681 acctcctggc tgatgatgcc gttccaggcg gggtatctga tgctgccctc ggcgctgccc
     7741 ttcagctgct cgatgtgctg ccactcctcg atggggttgg tctcgtcctt cagggcgatc
     7801 atctccttgc tgatggggtc gtaggcgtag tacctggaca ccagggcgaa gtagtggtcg
     7861 gggatggcgg tgatctggt
//
