LOCUS       sequence-403193-        6671 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses nuclear localized tdTomato under CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6671)
  AUTHORS   Feng
  TITLE     Feng Lab AAV plasmid
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6671)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-CAG-2xNLS-tdTomato-WPRE-hGHpolyA
FEATURES             Location/Qualifiers
     source          1..6671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        860..1163
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1165..1448
                     /label=chicken beta-actin promoter
     primer_bind     1778..1800
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1862..1881
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1924..1933
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1939..1959
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      1966..1975
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1972..3393
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNF
                     EDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLK
                     GEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERS
                     EGRHHLFLYGMDELYK"
     CDS             3400..3420
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    3448..4036
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3501..3521)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3919..3930)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4068..4544
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4267..4286)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4584..4724
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4584..4713
                     /label=AAV2 ITR
     rep_origin      4799..5254
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4886..4905)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5096..5117
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5271..5290)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5390..5412
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5450..5468)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5536..5640
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5641..5709,5710..6501)
                     /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(5859..5878)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gcgtaacact agttattaat agtaatcaat tacggggtca ttagttcata
      841 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      901 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      961 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     1021 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     1081 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     1141 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
     1201 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
     1261 cgatgggggc gggggggggg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg
     1321 aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc
     1381 gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc
     1441 ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc
     1501 gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc
     1561 cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc
     1621 tgcgtgaaag ccttgagggg ctccgggagc gccggcagga aggaaatggg cggggagggc
     1681 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1741 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1801 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1861 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattgcg gcccaacggt
     1921 accgccacca tgggaagccc aaagaagaag cgtaaggtag gatccgccac catggtgagc
     1981 aagggcgagg aggtcatcaa agagttcatg cgcttcaagg tgcgcatgga gggctccatg
     2041 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     2101 accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc
     2161 ccccagttca tgtacggctc caaggcgtac gtgaagcacc ccgccgacat ccccgattac
     2221 aagaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     2281 ggtctggtga ccgtgaccca ggactcctcc ctgcaggacg gcacgctgat ctacaaggtt
     2341 aagatgcgcg gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gaccatgggc
     2401 tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg cgagatccac
     2461 caggccctga agctgaagga cggcggccac tacctggtgg agttcaagac catctacatg
     2521 gccaagaagc ccgtgcaact gcccggctac tactacgtgg acaccaagct ggacatcacc
     2581 tcccacaacg aggactacac catcgtggaa cagtacgagc gctccgaggg ccgccaccac
     2641 ctgttcctgg ggcatggcac cggcagcacc ggcagcggca gctccggcac cgcctcctcc
     2701 gaggacaaca acatggccgt catcaaagag ttcatgcgct tcaaggtgcg catggagggc
     2761 tccatgaacg gccacgagtt cgagatcgag ggcgagggcc gcccctacga gggcacccag
     2821 accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc
     2881 ccccagttca tgtacggctc caaggcgtac gtgaagcacc ccgccgacat ccccgattac
     2941 aagaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     3001 ggtctggtga ccgtgaccca ggactcctcc ctgcaggacg gcacgctgat ctacaaggtt
     3061 aagatgcgcg gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gaccatgggc
     3121 tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg cgagatccac
     3181 caggccctga agctgaagga cggcggccac tacctggtgg agttcaagac catctacatg
     3241 gccaagaagc ccgtgcaact gcccggctac tactacgtgg acaccaagct ggacatcacc
     3301 tcccacaacg aggactacac catcgtggaa cagtacgagc gctccgaggg ccgccaccac
     3361 ctgttcctgt acggcatgga cgagctgtac aagggaagcc ccaagaaaaa gcggaaggtg
     3421 tgagaattcg atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga
     3481 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     3541 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     3601 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     3661 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     3721 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     3781 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     3841 aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg
     3901 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     3961 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     4021 tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct
     4081 gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc
     4141 ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta
     4201 tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct
     4261 gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct
     4321 ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg
     4381 catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat
     4441 tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat
     4501 tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa
     4561 ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     4621 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     4681 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     4741 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     4801 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4861 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4921 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4981 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     5041 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5101 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     5161 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5221 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     5281 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     5341 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     5401 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     5461 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     5521 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5581 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5641 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5701 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5761 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5821 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5881 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5941 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     6001 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6061 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6121 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     6181 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     6241 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     6301 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     6361 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6421 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6481 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6541 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6601 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6661 aaaggatctt c
//
