LOCUS       sequence-316520-        5402 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Cell morphology marker construct. Contains a red fluorescent protein
            mRuby3 and six FLAG tags..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5402)
  AUTHORS   Linghu C, Johnson SL, Valdes PA, Shemesh OA, Park WM, Park D, 
            Piatkevich KD, Wassie AT, Liu Y, An B, Barnes SA, Celiker OT, Yao 
            CC, Yu CJ, Wang R, Adamala KP, Bear MF, Keating AE, Boyden ES
  TITLE     Spatial Multiplexing of Fluorescent Reporters for Imaging Signaling 
            Network Dynamics.
  JOURNAL   Cell. 2020 Nov 17. pii: S0092-8674(20)31399-4. doi: 
            10.1016/j.cell.2020.10.035.
  PUBMED    33232692
REFERENCE   2  (bases 1 to 5402)
  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: "Cell. 2020 
            Nov 17. pii: S0092-8674(20)31399-4. doi: 
            10.1016/j.cell.2020.10.035."
COMMENT     Sequence Label: pAAV-hSyn-mRuby3-6xFLAG
FEATURES             Location/Qualifiers
     source          1..5402
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   58..187
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     polyA_signal    243..291
                     /note="synthetic polyadenylation signal"
     misc_feature    305..396
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     345..364
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        432..879
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      888..897
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             897..920
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             924..947
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             951..974
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             990..1697
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTDI
                     ALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAVA
                     KYSNLGGGMDELYK"
     CDS             1713..1736
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1740..1763
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1767..1790
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    1806..2394
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1859..1879)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2277..2288)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2427..2444)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2433..2640
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2649..2778
                     /label=AAV2 ITR
     rep_origin      2887..3342
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2974..2993)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3184..3205
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3409..3428)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3528..3550
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3588..3606)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3674..3778
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3779..3847,3848..4639)
                     /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(3997..4016)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4810..5398
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5299..5318
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct
       61 gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt
      121 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
      181 ggttcctgcg gccgcacgcg ttctagaaat attaaggtac gggaggtact tggagcggcc
      241 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
      301 tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg
      361 tccccagtgc aagtgcaggt gccagaacat ttctctattc tagactcgag ctgcagaggg
      421 ccctgcgtat gagtgcaagt gggttttagg accaggatga ggcggggtgg gggtgcctac
      481 ctgacgaccg accccgaccc actggacaag cacccaaccc ccattcccca aattgcgcat
      541 cccctatcag agagggggag gggaaacagg atgcggcgag gcgcgtgcgc actgccagct
      601 tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc
      661 actgaaggcg cgctgacgtc actcgccggt cccccgcaaa ctccccttcc cggccacctt
      721 ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg cgagataggg
      781 gggcacgggc gcgaccatct gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg
      841 gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt cgaattcgcc accatggatt
      901 acaaagatga cgatgacaaa ggcgactaca aggacgatga cgacaaagga gactacaagg
      961 atgatgacga caaaggtgga ggaggaggtg tgtctaaggg cgaagagctg atcaaggaaa
     1021 atatgcgtat gaaggtggtc atggaaggtt cggtcaacgg ccaccaattc aaatgcacag
     1081 gtgaaggaga aggcagaccg tacgagggag tgcaaaccat gaggatcaaa gtcatcgagg
     1141 gaggacccct gccatttgcc tttgacattc ttgccacgtc gttcatgtat ggcagccgta
     1201 cctttatcaa gtacccggcc gacatccctg atttctttaa acagtccttt cctgagggtt
     1261 ttacttggga aagagttacg agatacgaag atggtggagt cgtcaccgtc acgcaggaca
     1321 ccagccttga ggatggcgag ctcgtctaca acgtcaaggt cagaggggta aactttccct
     1381 ccaatggtcc cgtgatgcag aagaagacca agggttggga gcctaataca gagatgatgt
     1441 atccagcaga tggtggtctg agaggataca ctgacatcgc actgaaagtt gatggtggtg
     1501 gccatctgca ctgcaacttc gtgacaactt acaggtcaaa aaagaccgtc gggaacatca
     1561 agatgcccgg tgtccatgcc gttgatcacc gcctggaaag gatcgaggag agtgacaatg
     1621 aaacctacgt agtgcaaaga gaagtggcag ttgccaaata cagcaacctt ggtggtggca
     1681 tggacgagct gtacaaggga ggtggaggag gagattacaa agatgatgat gataagggag
     1741 attacaagga tgatgacgat aagggagact acaaggatga tgatgacaag taaaagctta
     1801 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     1861 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     1921 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     1981 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     2041 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     2101 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     2161 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     2221 tgctcgcctg tgttgccacc tggattctgc gcgggacgcc cttctgctac gtcccttcgg
     2281 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     2341 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     2401 accgtcgact agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2461 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2521 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2581 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     2641 gagatctgag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     2701 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     2761 gagcgagcga gcgcgcagag agggagtggc cccccccccc cccccccccc tgcagcctgg
     2821 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     2881 gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2941 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3001 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3061 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3121 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3181 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3241 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3301 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct
     3361 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
     3421 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     3481 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     3541 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     3601 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     3661 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3721 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3781 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3841 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3901 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3961 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4021 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4081 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4141 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4201 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4261 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4321 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4381 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4441 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4501 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4561 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4621 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4681 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4741 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4801 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4861 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4921 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4981 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5041 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5101 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5161 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5221 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5281 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5341 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5401 cg
//
