LOCUS       sequence-355469-        6710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PA-Flp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6710)
  AUTHORS   Jung H, Kim SW, Kim M, Hong J, Yu D, Kim JH, Lee Y, Kim S, Woo D, 
            Shin HS, Park BO, Do Heo W
  TITLE     Noninvasive optical activation of Flp recombinase for genetic 
            manipulation in deep mouse brain regions.
  JOURNAL   Nat Commun. 2019 Jan 18;10(1):314. doi: 10.1038/s41467-018-08282-8.
  PUBMED    30659191
REFERENCE   2  (bases 1 to 6710)
  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"; journalName: "Nat 
            Commun."; date: "2019-01-18"; volume: "10(1):314. doi"; pages: " 
            10.1038/s41467-018-08282-8"
COMMENT     Sequence Label: pAAV-EF1a-mCherry-P2A-NLSFlpN-nMagHigh
FEATURES             Location/Qualifiers
     source          1..6710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   2..131
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        232..1410
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          463..1401
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1358..1378
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1444..2151
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1589..1607)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1857..1876
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2161..2217
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2218..2238
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    2839..3426
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2892..2912)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3309..3320)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3458..3934
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3657..3676)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3974..4114
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3974..4103
                     /label=AAV2 ITR
     rep_origin      4189..4644
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4276..4295)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4486..4507
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4661..4680)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4780..4802
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4840..4858)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4926..5030
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5031..5099,5100..5891)
                     /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(5249..5268)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6062..6650
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6551..6570
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt
       61 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
      121 taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt ttaaacagag
      181 aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccggt
      241 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
      301 ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
      361 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
      421 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt
      481 tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc
      541 tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg
      601 aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc
      661 tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag
      721 tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt
      781 cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc
      841 gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca
      901 ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg
      961 ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga
     1021 gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc
     1081 tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc
     1141 gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg
     1201 agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc
     1261 cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg
     1321 gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa
     1381 agtttttttc ttccatttca ggtgtcgtga ggtaccggat cctctagagt cgactccgga
     1441 accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     1501 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     1561 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     1621 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     1681 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     1741 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     1801 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     1861 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     1921 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     1981 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     2041 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     2101 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gggatccggc
     2161 gcaacaaact tctctctgct gaaacaagcc ggagatgtcg aagagaatcc tggaccgccc
     2221 aagaagaaga ggaaggtgag ccagttcgac atcctgtgca agaccccccc caaggtgctg
     2281 gtgcggcagt tcgtggagag attcgagagg cccagcggtg gaggcggttc aggcggaggt
     2341 ggcagcggcg gtggcggatc gcacaccctg tacgcccccg gcggctacga catcatgggc
     2401 tacctggacc agatcggcaa cagacccaac ccccaggtgg agctgggccc cgtggacacc
     2461 agctgcgccc tgatcctgtg cgacctgaag cagaaggaca cccccatcgt gtacgccagc
     2521 gaggccttcc tgtatatgac cggctacagc aacgccgagg tgctgggcag aaactgcaga
     2581 ttcctgcaga gccccgacgg catggtgaag cccaagagca ccagaaagta cgtggacagc
     2641 aacaccatca acaccatcag aaaggccatc gacagaaacg ccgaggtgca ggtggaggtg
     2701 gtgaacttca agaagaacgg ccagagattc gtgaacttcc tgaccatcat ccccgtgaga
     2761 gacgagaccg gcgagtacag atacagcatg ggcttccagt gcgagaccga gtaagaattc
     2821 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2881 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2941 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3001 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3061 gctgacgcaa cccccactgg ttggggcatt gccaccactg tcagctcctt tccgggactt
     3121 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     3181 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     3241 cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct
     3301 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     3361 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     3421 ccccgcatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc
     3481 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat
     3541 aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag
     3601 gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta
     3661 ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg
     3721 ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac
     3781 caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct
     3841 ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt
     3901 acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg
     3961 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     4021 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     4081 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     4141 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     4201 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4261 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4321 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4381 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     4441 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4501 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     4561 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4621 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     4681 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4741 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4801 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4861 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4921 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4981 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5041 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5101 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5161 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5221 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5281 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5341 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5401 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5461 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5521 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5581 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5641 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5701 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5761 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5821 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5881 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5941 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6001 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6061 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6121 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6181 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6241 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6301 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6361 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6421 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6481 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6541 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6601 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6661 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
//
