LOCUS       sequence-261963-        7076 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent tdTomato fluorescent reporter for cell labeling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7076)
  AUTHORS   Sun YJ, Espinosa JS, Hoseini MS, Stryker MP
  TITLE     Experience-dependent structural plasticity at pre- and postsynaptic 
            sites of layer 2/3 cells in developing visual cortex.
  JOURNAL   Proc Natl Acad Sci U S A. 2019 Oct 22;116(43):21812-21820. doi: 
            10.1073/pnas.1914661116. Epub 2019 Oct 7.
  PUBMED    31591211
REFERENCE   2  (bases 1 to 7076)
  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"; doi: 
            "10.1073/pnas.1914661116"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2019-10-22- 22"; volume: "116"; issue: "43"; pages: 
            "21812-21820"
COMMENT     Sequence Label: pAAV-EF1a-DIO-tdTomato-WPRE
FEATURES             Location/Qualifiers
     source          1..7076
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1421..1430
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1486..1519
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    1570..1603
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1628..2216
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1681..1701)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2099..2110)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2248..2724
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2447..2466)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2764..2904
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2764..2893
                     /label=AAV2 ITR
     rep_origin      2979..3434
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3066..3085)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3276..3297
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3451..3470)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3570..3592
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3630..3648)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3716..3820
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3821..3889,3890..4681)
                     /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(4039..4058)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4852..5440
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5341..5360
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5502..5631
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        5732..6910
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5963..6901
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     6858..6878
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(6941..6974)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(7025..7058)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(7070..7076,1..1424))
                     /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
                     HLFLGHGIGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
ORIGIN
        1 tacagctcgt ccatgccgta caggaacagg tggtggcggc cctcggagcg ctcgtactgt
       61 tccacgatgg tgtagtcctc gttgtgggag gtgatgtcca gcttggtgtc cacgtagtag
      121 tagccgggca gttgcacggg cttcttggcc atgtagatgg tcttgaactc caccaggtag
      181 tggccgccgt ccttcagctt cagggcctgg tggatctcgc ccttcagcac gccgtcgcgg
      241 gggtacaggc gctcggtgga ggcctcccag cccatggtct tcttctgcat tacggggccg
      301 tcggggggga agttggtgcc gcgcatcttc accttgtaga tcagcgtgcc gtcctgcagg
      361 gaggagtcct gggtcacggt caccagaccg ccgtcctcga agttcatcac gcgctcccac
      421 ttgaagccct cggggaagga cagcttcttg taatcgggga tgtcggcggg gtgcttcacg
      481 tacgccttgg agccgtacat gaactggggg gacaggatgt cccaggcgaa gggcaggggg
      541 ccgcccttgg tcaccttcag cttggcggtc tgggtgccct cgtaggggcg gccctcgccc
      601 tcgccctcga tctcgaactc gtggccgttc atggagccct ccatgcgcac cttgaagcgc
      661 atgaactctt tgatgacggc catgttgttg tcctcggagg aggcggtgcc ggagctgccg
      721 ctgccggtgc tgccgatgcc atgccccagg aacaggtggt ggcggccctc ggagcgctcg
      781 tactgttcca cgatggtgta gtcctcgttg tgggaggtga tgtccagctt ggtgtccacg
      841 tagtagtagc cgggcagttg cacgggcttc ttggccatgt agatggtctt gaactccacc
      901 aggtagtggc cgccgtcctt cagcttcagg gcctggtgga tctcgccctt cagcacgccg
      961 tcgcgggggt acaggcgctc ggtggaggcc tcccagccca tggtcttctt ctgcattacg
     1021 gggccgtcgg gggggaagtt ggtgccgcgc atcttcacct tgtagatcag cgtgccgtcc
     1081 tgcagggagg agtcctgggt cacggtcacc agaccgccgt cctcgaagtt catcacgcgc
     1141 tcccacttga agccctcggg gaaggacagc ttcttgtaat cggggatgtc ggcggggtgc
     1201 ttcacgtacg ccttggagcc gtacatgaac tggggggaca ggatgtccca ggcgaagggc
     1261 agggggccgc ccttggtcac cttcagcttg gcggtctggg tgccctcgta ggggcggccc
     1321 tcgccctcgc cctcgatctc gaactcgtgg ccgttcatgg agccctccat gcgcaccttg
     1381 aagcgcatga actctttgat gacctcctcg cccttgctca ccatggtggc aagggcgaat
     1441 tccagcacac tggcggccgt tactagtgga tccgagctcg ctagcataac ttcgtataaa
     1501 gtatcctata cgaagttatt tgccttaacc cagaaattat cactgttatt ctttagaatg
     1561 gtgcaaagaa taacttcgta taatgtatgc tatacgaagt tatgaattcg atatcaagct
     1621 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     1681 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     1741 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     1801 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     1861 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     1921 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     1981 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     2041 gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     2101 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     2161 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     2221 ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct
     2281 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
     2341 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
     2401 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
     2461 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
     2521 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
     2581 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac
     2641 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
     2701 accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc
     2761 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     2821 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     2881 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     2941 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     3001 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     3061 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3121 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3181 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3241 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3301 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     3361 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3421 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3481 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3541 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3601 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     3661 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     3721 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3781 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3841 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3901 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3961 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4021 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4081 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4141 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4201 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4261 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4321 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4381 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4441 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4501 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4561 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4621 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4681 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4741 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4801 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4861 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4921 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4981 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5041 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5101 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5161 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5221 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5281 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5341 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5401 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5461 tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc agctgcgcgc
     5521 tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
     5581 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgca
     5641 cgcgtaagct ttgcaaagat ggataaagtt ttaaacagag aggaatcttt gcagctaatg
     5701 gaccttctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg
     5761 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta
     5821 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
     5881 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
     5941 cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt
     6001 tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt acgtgattct
     6061 tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc
     6121 cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat
     6181 ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt
     6241 ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga
     6301 tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca
     6361 gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta
     6421 gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc
     6481 ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc
     6541 cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga
     6601 gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac
     6661 ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc
     6721 tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt gggtggagac
     6781 tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt
     6841 tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttca
     6901 ggtgtcgtga ggtaccggat cctctagagt cgactccgga ataacttcgt ataggatact
     6961 ttatacgaag ttatgcagaa tggtagctgg attgtagctg ctattagcaa tatgaaacct
     7021 cttaataact tcgtatagca tacattatac gaagttatgg cgcgccctat tacttg
//
