LOCUS       sequence-355550-        6482 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FdCd.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6482)
  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 6482)
  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-hSynI-fDIO(TagBFP)-KoNLS-Cre(MtoL)
FEATURES             Location/Qualifiers
     source          1..6482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             648..668
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    680..713
                     /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             717..1427
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     protein_bind    complement(2457..2490)
                     /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)"
     regulatory      2501..2510
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2529..2538
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2611..3199
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2664..2684)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3082..3093)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3202..3218)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3203..3219)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3229..3705
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3428..3447)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3745..3885
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3745..3874
                     /label=AAV2 ITR
     rep_origin      3960..4415
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4047..4066)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4257..4278
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4432..4451)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4551..4573
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4611..4629)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4697..4801
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4802..4870,4871..5662)
                     /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(5020..5039)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5833..6421
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6322..6341
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggtgc caccatgccc aagaagaaga
      661 ggaaggtgct tcgtccggag aagttcctat tctctagaaa gtataggaac ttcaccatgg
      721 tgtctaaggg cgaagagctg attaaggaga acatgcacat gaagctgtac atggagggca
      781 ccgtggacaa ccatcacttc aagtgcacat ccgagggcga aggcaagccc tacgagggca
      841 cccagaccat gagaatcaag gtggtcgagg gcggccctct ccccttcgcc ttcgacatcc
      901 tggctactag cttcctctac ggcagcaaga ccttcatcaa ccacacccag ggcatccccg
      961 acttcttcaa gcagtccttc cctgagggct tcacatggga gagagtcacc acatacgaag
     1021 acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggctgc ctcatctaca
     1081 acgtcaagat cagaggggtg aacttcacat ccaacggccc tgtgatgcag aagaaaacac
     1141 tcggctggga ggccttcacc gagacgctgt accccgctga cggcggcctg gaaggcagaa
     1201 acgacatggc cctgaagctc gtgggcggga gccatctgat cgcaaacgcc aagaccacat
     1261 atagatccaa gaaacccgct aagaacctca agatgcctgg cgtctactat gtggactaca
     1321 gactggaaag aatcaaggag gccaacaacg agacctacgt cgagcagcac gaggtggcag
     1381 tggccagata ctgcgacctc cctagcaaac tggggcacaa gcttaattaa gaagttccta
     1441 ttcttcaaaa ggtataggaa cttcggcgcg ccttagtccc catcctcgag cagcctcacc
     1501 atggccccag tctcagagtc caggtttctg atgtagttca tcacaatgtt cacattggtc
     1561 cagccaccag cctgcatgat ttcagggatg gacacaccag ccctggccat gtccctggca
     1621 gcacccactc tggcagagtg gccagaccag gccaggtatc tctgcccaga gtcatccttg
     1681 gcaccataga tcaggcggtg ggtggcctca aagatccctt ccagggcccg ggtggacagt
     1741 tgggaggtgg cagaaggggc agccacacca ttctttctga cccggcagaa caggtagttg
     1801 ttggggtcat cagccacacc agacacagag atccatctct ccaccagctt ggtaaccccc
     1861 agggacaggg ccttctccac accagctgtg gacaccaggg tcttggtcct gccaatgtgg
     1921 atcagcattc tcccaccatc ggtgcgggag atgtccttca ctctgattct ggcaatttcg
     1981 gcaatgcgca gcagggtgtt gtaggcaatg cccaggaagg ccaggttcct gatgtcctgg
     2041 catctgtcag agttctccat cagggatctg acttggtcaa agtcagtgcg ttcaaaggcc
     2101 agggcctgct tggctctctc cccagcatcc acattctcct ttctgattct cctcatcacc
     2161 agggacacag cattggagtc agaagggcga ggcaggccag atctcctgtg cagcatgttg
     2221 agctggccca ggtgctgttg gatggtcttc acagccaggc ctctggcttg caggtacagg
     2281 aggtagtccc tcacatcctc aggttcagca gggaaccatt tcctgttgtt cagcttgcac
     2341 caggcagccc aggatctgca cacagacagg agcatcttcc aggtgtgttc agagaaggcc
     2401 tgcctgtccc tgaacaggtc cagcaggttc ttcctgactt catcagaggt gctagcgaag
     2461 ttcctatact ttctagagaa taggaacttc ttgccttaac ccatggtggc tcactgttat
     2521 tctttagacc atggtggcga atggtgcaaa gagaagttcc tatacctttt gaagaatagg
     2581 aacttcgaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     2641 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     2701 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     2761 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     2821 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     2881 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     2941 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     3001 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     3061 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     3121 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     3181 ctttgggccg cctccccgca tcgataccgt cgacctcgag agatctacgg gtggcatccc
     3241 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc
     3301 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
     3361 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
     3421 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc
     3481 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag
     3541 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata
     3601 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa
     3661 ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta
     3721 accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     3781 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     3841 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     3901 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     3961 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4021 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4081 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4141 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     4201 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4261 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     4321 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4381 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     4441 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     4501 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4561 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     4621 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4681 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4741 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4801 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4861 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4921 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4981 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     5041 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     5101 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5161 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5221 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5281 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5341 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     5401 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5461 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5521 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5581 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5641 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     5701 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5761 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5821 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5881 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5941 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6001 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6061 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6121 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6181 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6241 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6301 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6361 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6421 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6481 gt
//
