LOCUS       sequence-316521-        5633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cell morphology marker construct. Contains a near infrared 
            fluorescent protein miRFP and six FLAG tags..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5633)
  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 5633)
  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: "Cell. 2020 
            Nov 17. pii: S0092-8674(20)31399-4. doi: 
            10.1016/j.cell.2020.10.035."
COMMENT     Sequence Label: pAAV-hSyn-miRFP-6xFLAG
FEATURES             Location/Qualifiers
     source          1..5633
                     /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             1944..1967
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1971..1994
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1998..2021
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2037..2625
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2090..2110)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2508..2519)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2658..2675)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2664..2871
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2880..3009
                     /label=AAV2 ITR
     rep_origin      3118..3573
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3205..3224)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3415..3436
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3640..3659)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3759..3781
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3819..3837)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3905..4009
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4010..4078,4079..4870)
                     /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(4228..4247)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5041..5629
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5530..5549
                     /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 ggaggaggta tggtcgcagg acacgcaagt ggaagccccg
     1021 actttggaac cgccgatccg agcgactgtg agagagaaga aatccatctg gccggatcca
     1081 tccagccaca cggaacactg ctggtggtct ccgagcctga ccatagaatc attcaggcat
     1141 ctgccaacgc cgctgaattc ctgaatctgg gaagtgtgct gggagtcccc ctggctgaga
     1201 tcgacgggga tctgctgatc aagattctgc cccacctgga ccctaccgca gaaggaatgc
     1261 cagtggccgt caggtgccgc attggaaacc cctccacaga gtacgatgga ctgatgcatc
     1321 gaccacctga aggaggactg atcattgagc tggaacgggc tgggccaccc attgatctga
     1381 gcggaacact ggctcccgca ctggagcgaa ttcgaactgc aggatccctg agggctctgt
     1441 gcgacgatac tgcactgctg ttccagcagt gtaccggcta cgaccgagtg atggtctatc
     1501 ggttcgatga acaggggcac ggagaggtgt attcagaaat acatgtgact ggactggaga
     1561 gctactttgg caacaggtat cctagctccc tcgtgccaca gatggcccgg agactgtacg
     1621 aaagacagag ggtgcgcgtc ctggtggatg tctcttatca gcctgtgcca ctggagccaa
     1681 gactgagtcc cctgactggc agggacctgg atatgtcagg gtgttttctg cgctctatga
     1741 gtcctactca cctgcagttc ctgaagaata tgggcgtgcg ggcaacactg gtggtcagcc
     1801 tggtggtcgg aggcaaactg tgggggctgg tgatctgcca ccattatctg ccacgcttca
     1861 tccattttga gctgcgagca atttgtgagc tgctggccga agctatcgca acccgcatta
     1921 cagccctggg aggtggagga ggagattaca aagatgatga tgataaggga gattacaagg
     1981 atgatgacga taagggagac tacaaggatg atgatgacaa gtaaaagctt atcgataatc
     2041 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     2101 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     2161 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     2221 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     2281 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     2341 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     2401 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     2461 gtgttgccac ctggattctg cgcgggacgc ccttctgcta cgtcccttcg gccctcaatc
     2521 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     2581 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac
     2641 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2701 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2761 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2821 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg agagatctga
     2881 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     2941 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     3001 agcgcgcaga gagggagtgg cccccccccc cccccccccc ctgcagcctg gcgtaatagc
     3061 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg
     3121 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3181 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3241 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3301 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3361 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3421 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3481 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3541 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     3601 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     3661 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3721 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3781 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     3841 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     3901 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3961 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4021 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4081 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4141 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4201 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4261 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4321 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4381 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4441 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4501 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4561 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4621 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4681 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4741 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4801 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4861 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4921 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4981 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5041 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5101 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5161 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5221 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5281 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5341 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5401 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5461 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5521 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5581 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acg
//
