LOCUS       sequence-315053-        5646 bp    DNA     circular SYN 14-APR-2025
DEFINITION  nucleus marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5646)
  AUTHORS   Li LL, Klein FM, Li Greci L, Popinigis A, Freudenberg F, Courtney MJ
  TITLE     Resonance energy transfer sensitises and monitors in situ switching 
            of LOV2-based optogenetic actuators.
  JOURNAL   Nat Commun. 2020 Oct 9;11(1):5107. doi: 10.1038/s41467-020-18816-8.
  PUBMED    33037199
REFERENCE   2  (bases 1 to 5646)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2020-10-9"; volume: "11(1):5107. doi"; pages: " 
            10.1038/s41467-020-18816-8"
COMMENT     Sequence Label: AAV-Synapsin-miRFP670-3XNLS-WPRE
FEATURES             Location/Qualifiers
     source          1..5646
                     /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)"
     regulatory      652..661
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             658..1602
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 670 nm (Shcherbakova et al., 2016)"
                     /label=miRFP670
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     CDS             1660..1680
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    1699..1755
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    1775..2363
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1828..1848)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2246..2257)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2393..2869
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2592..2611)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2909..3049
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2909..3038
                     /label=AAV2 ITR
     rep_origin      3124..3579
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3211..3230)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3421..3442
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3596..3615)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3715..3737
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3775..3793)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3861..3965
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3966..4034,4035..4826)
                     /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(4184..4203)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4997..5585
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5486..5505
                     /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 gtacagctag cgctaccggt cgccaccatg
      661 gtagcaggtc atgcctctgg cagccccgca ttcgggaccg cctctcattc gaattgcgaa
      721 catgaagaga tccacctcgc cggctcgatc cagccgcatg gcgcgcttct ggtcgtcagc
      781 gaacatgatc atcgcgtcat ccaggccagc gccaacgccg cggaatttct gaatctcgga
      841 agcgtactcg gcgttccgct cgccgagatc gacggcgatc tgttgatcaa gatcctgccg
      901 catctcgatc ccaccgccga aggcatgccg gtcgcggtgc gctgccggat cggcaatccc
      961 tctacggagt actgcggtct gatgcatcgg cctccggaag gcgggctgat catcgaactc
     1021 gaacgtgccg gcccgtcgat cgatctgtca ggcacgctgg cgccggcgct ggagcggatc
     1081 cgcacggcgg gttcactgcg cgcgctgtgc gatgacaccg tgctgctgtt tcagcagtgc
     1141 accggctacg accgggtgat ggtgtatcgt ttcgatgagc aaggccacgg cctggtattc
     1201 tccgagtgcc atgtgcctgg gctcgaatcc tatttcggca accgctatcc gtcgtcgact
     1261 gtcccgcaga tggcgcggca gctgtacgtg cggcagcgcg tccgcgtgct ggtcgacgtc
     1321 acctatcagc cggtgccgct ggagccgcgg ctgtcgccgc tgaccgggcg cgatctcgac
     1381 atgtcgggct gcttcctgcg ctcgatgtcg ccgtgccatc tgcagttcct gaaggacatg
     1441 ggcgtgcgcg ccaccctggc ggtgtcgctg gtggtcggcg gcaagctgtg gggcctggtt
     1501 gtctgtcacc attatctgcc gcgcttcatc cgtttcgagc tgcgggcgat ctgcaaacgg
     1561 ctcgccgaaa ggatcgcgac gcggatcacc gcgcttgaga gcggcgagct gtacaagtcc
     1621 ggactcagat ccgaccccag aagaaagcgc aaggtggacc ccaagaagaa gcgcaaggtg
     1681 gaccccaaga agaagcgcag atctcgagct caagcttcga attctgcagt cgacggtacc
     1741 gcgggcccgg gatccgcggc cgctagataa ctgaaatcaa cctctggatt acaaaatttg
     1801 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     1861 tttaatgcct ttgtatcatg ctattgcttc ccgtatggcc ttcattttct cctccttgta
     1921 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     1981 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     2041 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     2101 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     2161 gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg
     2221 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     2281 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     2341 ctccctttgg gccgcctccc cgcatcgatt gtaccgcggg cccgggatct acgggtggca
     2401 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac
     2461 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa
     2521 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag
     2581 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc
     2641 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt
     2701 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac
     2761 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc
     2821 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta
     2881 ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     2941 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     3001 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     3061 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     3121 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3181 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3241 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3301 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     3361 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3421 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     3481 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     3541 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     3601 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     3661 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     3721 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     3781 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     3841 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3901 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3961 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4021 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4081 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4141 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4201 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4261 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4321 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4381 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4441 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4501 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4561 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4621 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4681 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4741 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4801 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4861 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4921 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4981 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5041 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5101 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5161 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5221 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5281 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5341 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5401 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5461 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5521 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5581 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5641 acatgt
//
