LOCUS       sequence-201189-        6595 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 6595)
  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 6595)
  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-Flpo
FEATURES             Location/Qualifiers
     source          1..6595
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   202..342
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   202..331
                     /label=AAV2 ITR
     rep_origin      417..872
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(504..523)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     714..735
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(889..908)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1008..1030
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1068..1086)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1154..1258
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1259..1327,1328..2119)
                     /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(1477..1496)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2290..2878
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2779..2798
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2940..3069
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        3170..4348
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3401..4339
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4296..4316
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    5661..6249
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5714..5734)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6132..6143)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    join(6281..6595,1..162)
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(6480..6499)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
ORIGIN
        1 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
       61 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
      121 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc
      181 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
      241 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
      301 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
      361 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
      421 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
      481 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
      541 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
      601 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
      661 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
      721 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
      781 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
      841 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
      901 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
      961 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     1021 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     1081 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     1141 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     1201 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     1261 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     1321 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     1381 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     1441 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     1501 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     1561 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     1621 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1681 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1741 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1801 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     1861 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     1921 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     1981 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     2041 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     2101 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     2161 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     2221 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     2281 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2341 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2401 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     2461 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2521 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2581 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2641 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2701 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2761 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2821 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2881 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtc
     2941 ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg
     3001 gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta
     3061 ggggttcctg cggccgcacg cgtaagcttt gcaaagatgg ataaagtttt aaacagagag
     3121 gaatctttgc agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc
     3181 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
     3241 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
     3301 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
     3361 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc
     3421 ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg
     3481 gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag
     3541 gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg
     3601 gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc
     3661 tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct
     3721 tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga
     3781 cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc
     3841 gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc
     3901 gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc
     3961 ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc
     4021 gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt
     4081 cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag
     4141 cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca
     4201 cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga
     4261 atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag
     4321 tttttttctt ccatttcagg tgtcgtgagg taccggatcc caccatgagc cagttcgaca
     4381 tcctgtgcaa gacccccccc aaggtgctgg tgcggcagtt cgtggagaga ttcgagaggc
     4441 ccagcggcga gaagatcgcc agctgtgccg ccgagctgac ctacctgtgc tggatgatca
     4501 cccacaacgg caccgccatc aagagggcca ccttcatgag ctacaacacc atcatcagca
     4561 acagcctgag cttcgacatc gtgaacaaga gcctgcagtt caagtacaag acccagaagg
     4621 ccaccatcct ggaggccagc ctgaagaagc tgatccccgc ctgggagttc accatcatcc
     4681 cttacaacgg ccagaagcac cagagcgaca tcaccgacat cgtgtccagc ctgcagctgc
     4741 agttcgagag cagcgaggag gccgacaagg gcaacagcca cagcaagaag atgctgaagg
     4801 ccctgctgtc cgagggcgag agcatctggg agatcaccga gaagatcctg aacagcttcg
     4861 agtacaccag caggttcacc aagaccaaga ccctgtacca gttcctgttc ctggccacat
     4921 tcatcaactg cggcaggttc agcgacatca agaacgtgga ccccaagagc ttcaagctgg
     4981 tgcagaacaa gtacctgggc gtgatcattc agtgcctggt gaccgagacc aagacaagcg
     5041 tgtccaggca catctacttt ttcagcgcca gaggcaggat cgaccccctg gtgtacctgg
     5101 acgagttcct gaggaacagc gagcccgtgc tgaagagagt gaacaggacc ggcaacagca
     5161 gcagcaacaa gcaggagtac cagctgctga aggacaacct ggtgcgcagc tacaacaagg
     5221 ccctgaagaa gaacgccccc taccccatct tcgctatcaa gaacggccct aagagccaca
     5281 tcggcaggca cctgatgacc agctttctga gcatgaaggg cctgaccgag ctgacaaacg
     5341 tggtgggcaa ctggagcgac aagagggcct ccgccgtggc caggaccacc tacacccacc
     5401 agatcaccgc catccccgac cactacttcg ccctggtgtc caggtactac gcctacgacc
     5461 ccatcagcaa ggagatgatc gccctgaagg acgagaccaa ccccatcgag gagtggcagc
     5521 acatcgagca gctgaagggc agcgccgagg gcagcatcag ataccccgcc tggaacggca
     5581 tcatcagcca ggaggtgctg gactacctga gcagctacat caacaggcgg atctgagaat
     5641 tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg
     5701 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     5761 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     5821 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     5881 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     5941 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     6001 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     6061 cgtcctttcc ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct
     6121 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     6181 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     6241 cctccccgca tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc
     6301 ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct
     6361 aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg
     6421 gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt
     6481 ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc
     6541 ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggca
//
