LOCUS       sequence-237805-        6519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent TdTomato expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6519)
  TITLE     AAV expression plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6519)
  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            "Unpublished"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-Flex-Rev-TdTomato
FEATURES             Location/Qualifiers
     source          1..6519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..278
                     /label=chicken beta-actin promoter
     primer_bind     557..579
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     641..660
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    695..728
                     /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    791..824
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      2296..2305
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2344..2377)
                     /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(2448..2481)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2523..3111
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2576..2596)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2994..3005)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3154..3275
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3165..3184)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3219..3238
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3437..3566
                     /label=AAV2 ITR
     rep_origin      complement(3803..4391)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3883..3902)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4562..5353,5354..5422))
                     /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     5185..5204
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5423..5527)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5553..6008)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5690..5711)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5902..5921
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6080..6209
                     /label=AAV2 ITR
ORIGIN
        1 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
       61 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg
      121 ggcgcgcgcc rggsggggsg gggsggggsg rggggsgggg sggggsgagg cggagaggtg
      181 cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc
      241 ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc
      301 gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt
      361 tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg
      421 tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg
      481 gccctttgtg cggggggagc ggctcggggc tgtccgcggg gggacggctg ccttcggggg
      541 ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct
      601 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg
      661 ctgtctcatc attttggcaa agaattggat ccccataact tcgtataaag tatcctatac
      721 gaagttatat caaaatagga agaccaatgc ttcaccatcg acccgaattg ccaagcatca
      781 ccatcgaccc ataacttcgt ataatgtatg ctatacgaag ttatactagt gctagagcta
      841 gctgatcact taaggtttaa acaccggttt atttatacag ttcgtccata ccatacagaa
      901 acagatgatg gcgaccttca gaacgctcgt actgttcaac aatagtataa tcctcgttgt
      961 gggaggtgat gtccagttta gtgtcaacgt agtaatagcc cggcagttgt acaggctttt
     1021 ttgccatgta gatagttttg aactcaacca gatagtgacc accatccttc agcttcaggg
     1081 cctggtgaat ctcgcctttc agcacaccgt cgcgcgggta cagacgttcg gtgctagctt
     1141 cccagcccat agttttcttc tgcattactg gaccgtcagg cgggaagtta gtgccacgca
     1201 ttttcacttt gtaaatcaga gtaccgtctt gcaggctgct gtcctgcgta acagtgacca
     1261 ggccgccatc ttcgaaattc ataacacgtt cccatttgaa gccctctggg aaggacagct
     1321 ttttgtaatc cgggatgtct gccgggtgtt ttacgtaggc tttggaaccg tacatgaact
     1381 gcggagacag gatatcccac gcaaaaggca gtgggccacc tttggtaact ttcagctttg
     1441 cggtctgagt gccctcgtat gggcgaccct cgccctcacc ttcaatctcg aactcatggc
     1501 cgttcataga accttccata cgtactttga acctcatgaa ctctttgatc accgccatgt
     1561 tgttgtcttc ggaagacgca gtgccagagg aaccagaacc cgtagaacca gtaccgtggc
     1621 ccaggaacag gtggtgacgg ccctcgctac gttcatactg ttcgacgatg gtataatctt
     1681 cgttgtgaga cgtgatatcc agtttggtgt ccacgtagta gtaacctggc agttgaaccg
     1741 gctttttagc catatagatc gtcttaaatt caaccaggta gtgaccgccg tccttcagtt
     1801 tcagcgcctg gtggatctca cctttcagaa caccgtcacg cggatacagg cgctcggtag
     1861 atgcttccca tcccatcgtc tttttctgca tcaccgggcc atccggcgga aagttcgtac
     1921 cacgcatctt caccttgtag atcagggtgc catcttgcag agagctatcc tgggtgacgg
     1981 taaccaggcc accgtcctca aagttcatta cacgctccca cttgaaaccc tccgggaaag
     2041 acagcttctt gtagtccggg atatcggctg ggtgtttgac gtaggccttg gaaccataca
     2101 taaactgcgg ggacaggatg tcccacgcga acggcagagg accgcctttg gttactttca
     2161 gtttagcggt ctgcgtaccc tcgtacggac gaccttcgcc ttcgccctcg atctcaaact
     2221 catgaccgtt catgctaccc tccatgcgta ctttaaaacg cataaactct ttaatcacct
     2281 cttcaccctt gctaaccatg gtggcctcga gtcgcgaacg cgtggcgcgc cggtaccact
     2341 agtataactt cgtataggat actttatacg aagttatcat tgggattctt cctattttga
     2401 tccaagcatc accatcgacc ctctagtcca gatctcacca tcgacccata acttcgtata
     2461 gcatacatta tacgaagtta tgtccctcga agaggttcga attcgatatc aagcttatcg
     2521 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     2581 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     2641 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     2701 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     2761 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     2821 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     2881 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     2941 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     3001 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     3061 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc
     3121 gtcgacccgg gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa
     3181 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     3241 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     3301 tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat
     3361 gtggtaaaat cgataaggat cttcctagag catggctacg tagataagta gcatggcggg
     3421 ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3481 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     3541 gcctcagtga gcgagcgagc gcgcagctgc attaatgaat cggccaacgc gcggggagag
     3601 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3661 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3721 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3781 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3841 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3901 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3961 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4021 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4081 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4141 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4201 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4261 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4321 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4381 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4441 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4501 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4561 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4621 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4681 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4741 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4801 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4861 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4921 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4981 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5041 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5101 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5161 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5221 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5281 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5341 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5401 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5461 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5521 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     5581 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     5641 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     5701 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     5761 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     5821 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     5881 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     5941 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     6001 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     6061 ctcttcgcta ttacgccagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
     6121 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
     6181 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt
     6241 atctacgtag ccatgctcta ggaagagtac cattgacgtc aataatgacg tatgttccca
     6301 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     6361 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     6421 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     6481 ggcagtacat ctacgtatta gtcatcgcta ttaccatgg
//
