LOCUS       sequence-315043-        5310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  untargeted marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5310)
  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 5310)
  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: "2020-10-9"; volume: "11(1):5107. doi"; pages: " 
            10.1038/s41467-020-18816-8"
COMMENT     Sequence Label: AAV-Synapsin-mScarlet-C1-WPRE
FEATURES             Location/Qualifiers
     source          1..5310
                     /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..1353
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(791..809)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1059..1078
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1354..1419
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    1439..2027
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1492..1512)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1910..1921)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2057..2533
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2256..2275)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2573..2713
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2573..2702
                     /label=AAV2 ITR
     rep_origin      2788..3243
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2875..2894)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3085..3106
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3260..3279)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3379..3401
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3439..3457)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3525..3629
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3630..3698,3699..4490)
                     /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(3848..3867)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4661..5249
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5150..5169
                     /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 gtgagcaagg gcgaggcagt gatcaaggag ttcatgcggt tcaaggtgca catggagggc
      721 tccatgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
      781 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttctc ctgggacatc
      841 ctgtcccctc agttcatgta cggctccagg gccttcacca agcaccccgc cgacatcccc
      901 gactactata agcagtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
      961 gacggcggcg ccgtgaccgt gacccaggac acctccctgg aggacggcac cctgatctac
     1021 aaggtgaagc tccgcggcac caacttccct cctgacggcc ccgtaatgca gaagaagaca
     1081 atgggctggg aagcgtccac cgagcggttg taccccgagg acggcgtgct gaagggcgac
     1141 attaagatgg ccctgcgcct gaaggacggc ggccgctacc tggcggactt caagaccacc
     1201 tacaaggcca agaagcccgt gcagatgccc ggcgcctaca acgtcgaccg caagttggac
     1261 atcacctccc acaacgagga ctacaccgtg gtggaacagt acgaacgctc cgagggccgc
     1321 cactccaccg gcggcatgga cgagctgtac aagtccggac tcagatctcg agctcaagct
     1381 tcgaattctg cagtcgacgg taccgcgggc ccgggatccg cggccgctag ataactgaaa
     1441 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     1501 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     1561 ggccttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     1621 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     1681 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     1741 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     1801 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     1861 ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     1921 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     1981 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gattgtaccg
     2041 cgggcccggg atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     2101 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     2161 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     2221 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
     2281 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
     2341 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
     2401 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
     2461 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
     2521 cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc
     2581 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     2641 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     2701 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     2761 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     2821 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     2881 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2941 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3001 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3061 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     3121 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     3181 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     3241 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     3301 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     3361 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     3421 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     3481 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3541 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3601 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     3661 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3721 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3781 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     3841 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     3901 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     3961 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4021 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4081 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4141 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     4201 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4261 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4321 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4381 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4441 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     4501 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4561 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4621 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4681 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4741 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4801 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4861 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4921 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4981 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5041 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5101 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5161 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5221 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5281 tggccttttg ctggcctttt gctcacatgt
//
