LOCUS       sequence-380113-        6130 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express a bright monomeric red FP to label eucaryotic cell 
            nuclei. To be used in tissue clearing methods and other fluorescent 
            microscopy methods.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6130)
  AUTHORS   Wang Z, Romanski A, Mehra V, Wang Y, Brannigan M, Campbell BC, 
            Petsko GA, Tsoulfas P, Blackmore MG
  TITLE     Brain-wide analysis of the supraspinal connectome reveals anatomical
            correlates to functional recovery after spinal injury.
  JOURNAL   Elife. 2022 Jul 15;11. pii: 76254. doi: 10.7554/eLife.76254.
  PUBMED    35838234
REFERENCE   2  (bases 1 to 6130)
  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: "Elife."; 
            date: "2022-07-15"; volume: "11. pii: 76254. doi"; pages: " 
            10.7554/eLife.76254"
COMMENT     Sequence Label: pAAV2-UbC-3xNLS-mScarlet-I
FEATURES             Location/Qualifiers
     source          1..6130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(101..120)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      914..1502
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1403..1422
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1564..1693
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        1715..2114
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2684..2703
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     primer_bind     3400..3419
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3509..3529
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3533..3553
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3557..3577
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      3590..3599
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3596..4291
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(3729..3747)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3997..4016
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4440..4916
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4639..4658)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4956..5096
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4956..5085
                     /label=AAV2 ITR
     rep_origin      5171..5626
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5258..5277)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5468..5489
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5643..5662)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5762..5784
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5822..5840)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5908..6012
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6013..6081,6082..6130,1..743)
                     /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"
ORIGIN
        1 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
       61 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
      121 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
      181 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
      241 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
      301 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
      361 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
      421 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
      481 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
      541 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
      601 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
      661 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
      721 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
      781 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
      841 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
      901 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
      961 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1021 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     1081 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1141 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1201 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1261 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     1321 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     1381 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     1441 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     1501 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     1561 tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc
     1621 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     1681 actaggggtt cctgcggccg cacgcgttta aactggcctc cgcgccgggt tttggcgcct
     1741 cccgcgggcg cccccctcct cacggcgagc gctgccacgt cagacgaagg gcgcagcgag
     1801 cgtcctgatc cttccgcccg gacgctcagg acagcggccc gctgctcata agactcggcc
     1861 ttagaacccc agtatcagca gaaggacatt ttaggacggg acttgggtga ctctagggca
     1921 ctggttttct ttccagagag cggaacaggc gaggaaaagt agtcccttct cggcgattct
     1981 gcggagggat ctccgtgggg cggtgaacgc cgatgattat ataaggacgc gccgggtgtg
     2041 gcacagctag ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt
     2101 gatcgtcact tggtgagtag cgggctgctg ggctggccgg ggctttcgtg gccgccgggc
     2161 cgctcggtgg gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag
     2221 caaggttgcc ctgaactggg ggttgggggg agcgcagcaa aatggcggct gttcccgagt
     2281 cttgaataga accttcgcta atgcgggaaa gctcttattc gggtgagatg ggctggggca
     2341 ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt tgtcgtctgt
     2401 tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg
     2461 cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg gtggggccac
     2521 ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc gggcctaggg
     2581 taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata agtgaggcgt
     2641 cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct ccggttttga
     2701 actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg
     2761 taatcatttg ggtcaatatg taattttcag tgttagacta gtaaattgtc cgctaaattc
     2821 tggccgtttt tggctttttt gttagacgaa gctaaggcgc gcctgagaac ttcagggtga
     2881 gtttggggac ccttgattgt tctttctttt tcgctattgt aaaattcatg ttatatggag
     2941 ggggcaaagt tttcagggtg ttgtttagaa tgggaagatg tcccttgtat caccatgcat
     3001 ggaccctcat gataattttg tttctttcac tttctactct gttgacaacc attgtctcct
     3061 cttattttct tttcattttc tgtaactttt tcgttaaact ttagcttgca tttgtaacga
     3121 atctagacaa tcagggtata ttatattgta cttcagcaca gttttagaga acaattgtta
     3181 taattaaatg ataaggtaga atatttctgc atataaattc tggctggcgt ggaaatattc
     3241 ttattggtag aaacaactac atcctggtca tcatcctgcc tttctcttta tggttacaat
     3301 gatatacact gtttgagatg aggataaaat actctgagtc caaaccgggc ccctctgcta
     3361 accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc
     3421 tgtctcatca agcttggatc cgaattcgct agcgctaccg gactcagatc tcgagctcaa
     3481 gcttcgaatt cccaccatgg gatcagatcc aaaaaagaag agaaaggtag acccgaagaa
     3541 aaagcgtaag gtcgatccca aaaagaaacg gaaagtggga tcaccggtcg ccaccatggt
     3601 gagcaagggc gaggcagtga tcaaggagtt catgcggttc aaggtgcaca tggagggctc
     3661 catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     3721 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttctcct gggacatcct
     3781 gtcccctcag ttcatgtacg gctccagggc cttcatcaag caccccgccg acatccccga
     3841 ctactataag cagtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     3901 cggcggcgcc gtgaccgtga cccaggacac ctccctggag gacggcaccc tgatctacaa
     3961 ggtgaagctc cgcggcacca acttccctcc tgacggcccc gtaatgcaga agaagacaat
     4021 gggctgggaa gcgtccaccg agcggttgta ccccgaggac ggcgtgctga agggcgacat
     4081 taagatggcc ctgcgcctga aggacggcgg ccgctacctg gcggacttca agaccaccta
     4141 caaggccaag aagcccgtgc agatgcccgg cgcctacaac gtcgaccgca agttggacat
     4201 cacctcccac aacgaggact acaccgtggt ggaacagtac gaacgctccg agggccgcca
     4261 ctccaccggc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc
     4321 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     4381 aacctgaaac ataaaatgaa tgcaattgtt gttgttaacg gccatattgg ccaatctacg
     4441 ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag
     4501 tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct
     4561 tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa
     4621 cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc
     4681 tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt
     4741 tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg
     4801 gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt
     4861 ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga
     4921 ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     4981 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     5041 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     5101 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     5161 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     5221 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     5281 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     5341 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     5401 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     5461 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     5521 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     5581 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     5641 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     5701 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5761 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     5821 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5881 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5941 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     6001 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     6061 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     6121 cagttgggtg
//
