LOCUS       sequence-110619-        7648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of CRYS2PHR-SID4X.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7648)
  AUTHORS   Chen Z, Li W, Qiu F, Huang Q, Jiang Z, Ye J, Cheng P, Low C, Guo Y, 
            Yi X, Chen W, Yu Y, Han Y, Wu J, Jin S, Kong D, Huang J
  TITLE     Aspirin cooperates with p300 to activate the acetylation of H3K9 and
            promote FasL-mediated apoptosis of cancer stem-like cells in 
            colorectal cancer.
  JOURNAL   Theranostics. 2018 Aug 7;8(16):4447-4461. doi: 10.7150/thno.24284. 
            eCollection 2018.
  PUBMED    30214631
REFERENCE   2  (bases 1 to 7648)
  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: 
            "10.7150/thno.24284"; journalName: "Theranostics"; date: "2018-08-7-
            7"; volume: "8"; issue: "16"; pages: "4447-4461"
COMMENT     Sequence Label: pEF1a-CRY2PHR-SID4X
FEATURES             Location/Qualifiers
     source          1..7648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             2944..2964
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3364..3417
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    3775..4363
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3828..3848)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4246..4257)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4395..4871
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4594..4613)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4911..5051
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4911..5040
                     /label=AAV2 ITR
     rep_origin      5126..5581
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5213..5232)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5423..5444
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5598..5617)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5717..5739
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5777..5795)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5863..5967
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5968..6036,6037..6828)
                     /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(6186..6205)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6999..7587
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7488..7507
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc cgccaccatg
     1441 aagatggaca aaaagacgat tgtgtggttt agacgtgacc ttcgcattga ggacaaccct
     1501 gcgttggcag cggcagcgca tgaaggttcg gtattccctg tctttatctg gtgtcccgaa
     1561 gaggaggggc aattctatcc cggtcgcgcg tcccgatggt ggatgaaaca gtcgctcgca
     1621 catcttagcc agtcgttgaa ggcgctcggc tctgatctga cgctcatcaa aacacacaat
     1681 acaatttctg ccattctgga ttgcattagg gtgactggtg ccaccaaagt tgtgttcaat
     1741 cacctttatg atcccgtaag cctggtcagg gatcatacag taaaagagaa acttgtagag
     1801 aggggaatca gcgtccaatc gtacaatggg gacttgttgt atgaaccgtg ggaaatctat
     1861 tgtgagaaag gcaaaccttt cacttccttc aattcgtact ggaagaaatg cctcgacatg
     1921 tccatcgagt cagtcatgct gccgccacct tggagactta tgcccattac tgctgcggca
     1981 gaggcgatct gggcttgcag catcgaagaa cttggtctgg aaaacgaagc ggaaaaaccc
     2041 agtaacgcgc tcctgacgcg agcgtggtcg ccaggctggt cgaatgctga taagctcctt
     2101 aatgagttta tcgagaaaca gttgatcgac tatgcgaaga atagcaagaa ggtggtggga
     2161 aattccactt ccctcctgtc gccgtatctc cactttgggg agattagcgt gaggcatgtg
     2221 ttccaatgtg cgcggatgaa gcaaatcatt tgggctcgcg acaaaaactc tgaaggagag
     2281 gagtctgccg atttgttttt gagaggaatc ggccttaggg agtattcacg atacatctgc
     2341 ttcaactttc cgtttacaca tgaacaatcc ctcctcagcc acctccgctt ctttccgtgg
     2401 gacgcggacg tggacaaatt caaggcctgg cggcaggggc gcacgggata cccgttggtg
     2461 gatgcgggta tgagagagct gtgggcaacg ggttggatgc acaaccgtat tcgagtcatc
     2521 gttagtagct ttgccgttaa gttcctcttg ttgccctgga aatgggggat gaaatacttt
     2581 tgggacacgc ttctggatgc ggacttggag tgcgatattc ttgggtggca gtacatttcg
     2641 ggttccatcc ctgacgggca tgaacttgac cgattggata accctgcact gcaaggagca
     2701 aagtacgacc cggagggaga atacatcagg cagtggctcc cagagctggc ccgtctgccc
     2761 accgagtgga ttcaccaccc gtgggatgct cccctgaccg tccttaaagc ctcaggggtc
     2821 gaattgggga cgaattacgc gaaacccatc gtcgatattg acaccgcacg agaattgctg
     2881 gcgaaagcca tctcaagaac acgcgaagcg cagattatga tcggagcagc gccggctagc
     2941 agccccaaga agaagagaaa ggtggaggcc agcatgaaca tccagatgct gctggaggcc
     3001 gctgactacc tggaacggag agagcgcgaa gccgagcacg gatatgcttc aatgctgccc
     3061 ggaagcggca tgaatattca gatgctgctg gaggctgctg attacctgga aaggcgcgaa
     3121 cgggaggccg aacatggcta tgcttccatg ctgcctgggt ctggaatgaa tatccaaatg
     3181 ctgctggagg cagccgatta cctggaacgg agagaaagag aagccgagca cggatacgcc
     3241 agcatgctgc caggcagcgg gatgaacata caaatgctgc tggaggctgc cgattacctg
     3301 gagaggcgcg agagagaagc tgaacatggc tatgcctcta tgctgccccc atggggcagt
     3361 ggagagggca gaggaagtct gctaacatgc ggtgacgtcg aggagaatcc tggcccaatc
     3421 gagaagtcct tcgtaataac agatccaaga ctgcctgatt accccatcat ttttgccagc
     3481 gacggttttc tggagctcac cgaatattcc agggaggaaa tcatggggag aaacgccagg
     3541 ttcctgcaag ggcctgagac agaccaggcc accgttcaga agattcgcga tgctatccgc
     3601 gatcagaggg aaaccactgt gcagttaata aactacacta agtcaggcaa aaagttctgg
     3661 aatctgcttc atctccaacc tgtccgggat cgaaagggag ggctgcaata cttcatcggt
     3721 gtgcagctgg tgggaagtga tcatgtgtaa gaattcgata tcaagcttat cgataatcaa
     3781 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     3841 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     3901 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     3961 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     4021 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     4081 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     4141 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat
     4201 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     4261 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     4321 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg
     4381 ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg
     4441 aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt
     4501 ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc
     4561 aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag
     4621 tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc
     4681 agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt
     4741 tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg
     4801 tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct
     4861 tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct
     4921 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     4981 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     5041 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     5101 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5161 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     5221 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     5281 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     5341 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5401 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     5461 atctcgggct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     5521 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     5581 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     5641 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     5701 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     5761 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     5821 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5881 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5941 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6001 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6061 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6121 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6181 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6241 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6301 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6361 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6421 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6481 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6541 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6601 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6661 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6721 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6781 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6841 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6901 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6961 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7021 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7081 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7141 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7201 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7261 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7321 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7381 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7441 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7501 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7561 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7621 gccttttgct ggccttttgc tcacatgt
//
