LOCUS       sequence-324113-        7226 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid expressing nuclear-localized FusionRed fluorophore.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7226)
  AUTHORS   Wang SK, Lapan SW, Hong CM, Krause TB, Cepko CL
  TITLE     In Situ Detection of Adeno-associated Viral Vector Genomes with 
            SABER-FISH.
  JOURNAL   Mol Ther Methods Clin Dev. 2020 Oct 10;19:376-386. doi: 
            10.1016/j.omtm.2020.10.003. eCollection 2020 Dec 11.
  PUBMED    33209963
REFERENCE   2  (bases 1 to 7226)
  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: "Mol Ther 
            Methods Clin Dev."; date: "2020-10-10"; volume: "19:376-386. doi"; 
            pages: " 10.1016/j.omtm.2020.10.003. eCollection 2020 Dec 11"
COMMENT     Sequence Label: AAV-CMV-H2B-FusionRed
FEATURES             Location/Qualifiers
     source          1..7226
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   9..138
                     /label=AAV2 ITR
     primer_bind     complement(194..213)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        315..694
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        695..898
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     849..869
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     895..919
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1012..1487
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     CDS             1543..1920
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      1933..1942
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1939..2637
                     /codon_start=1
                     /product="monomeric red fluorescent protein optimized for 
                     use as a fusion tag (Shemiakina et al., 2012)"
                     /label=FusionRed
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMPMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFMYGSRTFIKHPPGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKVRGVNFPANGPVMQKKTLGWEASTETMYPADGGLEGACDMAL
                     KLVGGGHLICNLETTYRSKKPATNLKMPGVYNVDHRLERIKEADDETYVEQHEVAVARY
                     STGGAGDGGK"
     primer_bind     complement(2655..2671)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2656..2672)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2765..3159
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     repeat_region   3514..3643
                     /label=AAV2 ITR
     rep_origin      4014..4527
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4101..4120)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4311..4332
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5009..5028)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5128..5150
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5188..5206)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5274..5378
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5379..5447,5448..6239)
                     /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(5597..5616)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6410..6998
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6899..6918
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatgcagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg
       61 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
      121 ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc
      181 catgctctag cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atgagcttgg
      241 cccattgcat acgttgtatc catatcataa tatgtacatt tatattggct catgtccaac
      301 attaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
      361 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      421 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      481 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      541 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      601 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      661 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      721 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      781 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      841 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
      901 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag
      961 acaccgggac cgatccagcc tcccctcgaa gctgatcctg agaacttcag ggtgagtcta
     1021 tgggaccctt gatgttttct ttccccttct tttctatggt taagttcatg tcataggaag
     1081 gggagaagta acagggtaca catattgacc aaatcagggt aattttgcat ttgtaatttt
     1141 aaaaaatgct ttcttctttt aatatacttt tttgtttatc ttatttctaa tactttccct
     1201 aatctctttc tttcagggca ataatgatac aatgtatcat gcctctttgc accattctaa
     1261 agaataacag tgataatttc tgggttaagg caatagcaat atttctgcat ataaatattt
     1321 ctgcatataa attgtaactg atgtaagagg tttcatattg ctaatagcag ctacaatcca
     1381 gctaccattc tgcttttatt ttatggttgg gataaggctg gattattctg agtccaagct
     1441 aggccctttt gctaatcatg ttcatacctc ttatcttcct cccacagctc ctgggcaacg
     1501 tgagctcaag cttcgaattc tgcagtcgac ggtaccgcca ccatgccaga gccagcgaag
     1561 tctgctcccg ccccgaaaaa gggctccaag aaggcggtga ctaaggcgca gaagaaaggc
     1621 ggcaagaagc gcaagcgcag ccgcaaggag agctattcca tctatgtgta caaggttctg
     1681 aagcaggtcc accctgacac cggcatttcg tccaaggcca tgggcatcat gaattcgttt
     1741 gtgaacgaca ttttcgagcg catcgcaggt gaggcttccc gcctggcgca ttacaacaag
     1801 cgctcgacca tcacctccag ggagatccag acggccgtgc gcctgctgct gcctggggag
     1861 ttggccaagc acgccgtgtc cgagggtact aaggccatca ccaagtacac cagcgctaag
     1921 gatccaccgg tcgccaccat ggtgagcgag ctgattaagg agaacatgcc catgaagctg
     1981 tacatggagg gcaccgtgaa caaccaccac ttcaagtgca catccgaggg cgaaggcaag
     2041 ccctacgagg gcacccagac catgagaatc aaggtcgtcg agggcggccc tctccccttc
     2101 gccttcgaca tcctggctac cagcttcatg tacggcagca gaaccttcat caagcaccct
     2161 ccgggcatcc ccgacttctt taagcagtcc ttccctgagg gcttcacatg ggagagagtc
     2221 accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc
     2281 tgcctcatct acaacgtcaa ggttagaggg gtgaacttcc cagccaacgg ccctgtgatg
     2341 cagaagaaaa cactcggctg ggaggcctcc accgagacga tgtaccccgc tgacggcggc
     2401 ctggaaggcg catgtgacat ggccctgaag ctcgtgggcg ggggccacct gatctgcaac
     2461 cttgagacca catacagatc caagaaaccc gctacgaacc tcaagatgcc cggcgtctac
     2521 aacgtggacc acagactgga aagaatcaag gaggccgacg atgagaccta cgtcgagcag
     2581 cacgaggtgg ctgtggccag atactctact ggtggcgctg gtgatggagg taaatgagcg
     2641 gccgccaagc ttatcgatac cgtcgacctc gagggcccag atctaattca ccccaccagt
     2701 gcaggctgcc tatcagaaag tggtggctgg tgtggctaat gccctggccc acaagtatca
     2761 ctaagctcgc tttcttgctg tccaatttct attaaaggtt cctttgttcc ctaagtccaa
     2821 ctactaaact gggggatatt atgaagggcc ttgagcatct ggattctgcc taataaaaaa
     2881 catttatttt cattgcaatg atgtatttaa attatttctg aatattttac taaaaaggga
     2941 atgtgggagg tcagtgcatt taaaacataa agaaatgaag agctagttca aaccttggga
     3001 aaatacacta tatcttaaac tccatgaaag aaggtgaggc tgcaaacagc taatgcacat
     3061 tggcaacagc ccctgatgcc tatgccttat tcatccctca gaaaaggatt caagtagagg
     3121 cttgatttgg aggttaaagt tttgctatgc tgtattttac attacttatt gttttagctg
     3181 tcctcatgaa tgtcttttca ctacccattt gcttatcctg catctctcag ccttgactcc
     3241 actcagttct cttgcttaga gataccacct ttcccctgaa gtgttccttc catgttttac
     3301 ggcgagatgg tttctcctcg cctggccact cagccttagt tgtctctgtt gtcttataga
     3361 ggtctacttg aagaaggaaa aacagggggc atggtttgac tgtcctgtga gcccttcttc
     3421 cctgcctccc ccactcacag tgacccggaa tccctcgaca tctagagcat ggctacgtag
     3481 ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca
     3541 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3601 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctggcgt aatagcgaag
     3661 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgattcc
     3721 gttgcaatgg ctggcggtaa tattgttctg gatattacca gcaaggccga tagtttgagt
     3781 tcttctactc aggcaagtga tgttattact aatcaaagaa gtattgcgac aacggttaat
     3841 ttgcgtgatg gacagactct tttactcggt ggcctcactg attataaaaa cacttctcag
     3901 gattctggcg taccgttcct gtctaaaatc cctttaatcg gcctcctgtt tagctcccgc
     3961 tctgattcta acgaggaaag cacgttatac gtgctcgtca aagcaaccat agtacgcgcc
     4021 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4081 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4141 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4201 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4261 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4321 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4381 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4441 ttttaacaaa atattaacgc ttacaattta aatatttgct tatacaatct tcctgttttt
     4501 ggggcttttc tgattatcaa ccggggtaca tatgattgac atgctagttt tacgattacc
     4561 gttcatcgat tctcttgttt gctccagact ctcaggcaat gacctgatag cctttgtaga
     4621 gacctctcaa aaatagctac cctctccggc atgaatttat cagctagaac ggttgaatat
     4681 catattgatg gtgatttgac tgtctccggc ctttctcacc cgtttgaatc tttacctaca
     4741 cattactcag gcattgcatt taaaatatat gagggttcta aaaattttta tccttgcgtt
     4801 gaaataaagg cttctcccgc aaaagtatta cagggtcata atgtttttgg tacaaccgat
     4861 ttagctttat gctctgaggc tttattgctt aattttgcta attctttgcc ttgcctgtat
     4921 gatttattgg atgttggaat cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4981 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5041 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     5101 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     5161 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     5221 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5281 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5341 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5401 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5461 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5521 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5581 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5641 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5701 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5761 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5821 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5881 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5941 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     6001 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6061 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6121 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6181 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6241 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6301 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6361 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6421 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6481 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6541 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6601 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6661 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6721 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6781 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6841 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6901 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6961 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     7021 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     7081 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     7141 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     7201 ctctccccgc gcgttggccg attcat
//
