LOCUS       Exported                6860 bp ds-DNA     circular SYN 16-MAR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6860)
  AUTHORS   SnapGene License
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 20, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6860
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1493..3561
                     /organism="Homo sapiens"
                     /chromosome="5"
                     /map="5q22.2"
                     /mol_type="mRNA"
                     /db_xref="taxon:9606"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     primer_bind     141..190
                     /label=F-D1
     primer_bind     147..194
                     /label=F-Mlu-D2
     primer_bind     147..161
                     /label=3'-pcalm-ampF
     primer_bind     150..168
                     /label=P-Calm1-F
     primer_bind     151..200
                     /label=F-D2
     misc_feature    156..276
                     /label=P Calm1
     misc_feature    156..165
                     /label=D1
     primer_bind     161..210
                     /label=F-D3
     primer_bind     complement(161..185)
                     /label=R-D4
     misc_feature    166..175
                     /label=D2
     primer_bind     171..220
                     /label=F-D4
     primer_bind     complement(171..195)
                     /label=R-D5
     misc_feature    176..185
                     /label=D3
     primer_bind     181..230
                     /label=F-D5
     primer_bind     complement(181..205)
                     /label=R-D6
     misc_feature    186..195
                     /label=D4
     primer_bind     191..240
                     /label=F-D6
     primer_bind     complement(191..215)
                     /label=r-d7
     misc_feature    196..205
                     /label=D5
     primer_bind     200..250
                     /label=F-D7
     primer_bind     complement(201..225)
                     /label=R-D8
     misc_feature    206..215
                     /label=D6
     primer_bind     211..260
                     /label=F-D8
     primer_bind     complement(215..236)
                     /label=R-D9
     misc_feature    216..225
                     /label=D7
     primer_bind     222..271
                     /label=F-D9
     primer_bind     complement(222..246)
                     /label=R-D10
     misc_feature    226..235
                     /label=D8
     primer_bind     complement(232..256)
                     /label=R-D11
     misc_feature    236
                     /label=+1
     misc_feature    237..246
                     /label=D9
     primer_bind     complement(242..266)
                     /label=R-D12
     misc_feature    247..256
                     /label=D10
     misc_feature    257..266
                     /label=D11
     primer_bind     complement(260..287)
                     /label=2B-1R
     primer_bind     complement(261..276)
                     /label=P-Calm1-R
     misc_feature    267..276
                     /label=D12
     primer_bind     271..296
                     /label=2B-2F
     primer_bind     complement(277..298)
                     /label=Primer 1
     misc_feature    277..282
                     /label=BsiWI
     primer_bind     283..302
                     /label=Primer 5
     CDS             288..1318
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     misc_feature    548
                     /label=silent mut
     primer_bind     complement(616..650)
                     /label=R-Cre-R119STOP
     primer_bind     complement(632..651)
                     /label=R-CRE-L115-BamHI
     primer_bind     636..670
                     /label=F-Cre-R119STOP
     primer_bind     1093..1116
                     /label=F-Seq-Cre-3F1
     misc_feature    1093..1098
                     /label=ClaI
     misc_feature    1224
                     /label=mut
     primer_bind     1262..1279
                     /label=cre-SF
     primer_bind     complement(1296..1318)
                     /label=R-T5-Cre
     intron          1320..1360
                     /label=Splicing donor
                     /note="Splicing donor"
     primer_bind     complement(1338..1367)
                     /label=2B-2R
     primer_bind     complement(1341..1385)
                     /label=Primer 7
     primer_bind     complement(1341..1385)
                     /label=Primer 6
     primer_bind     1350..1387
                     /label=2B-3F
     primer_bind     complement(1356..1380)
                     /label=Primer 3
     primer_bind     1367..1401
                     /label=Primer 8
     primer_bind     1367..1401
                     /label=Primer 2
     primer_bind     1367..1401
                     /label=F-T5-JT+2272
     protein_bind    1367..1400
                     /label=loxJTZ17?2272?
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    1380..1387
                     /label=2272
     intron          1401..1492
                     /label=Splicing acceptor
                     /note="Splicing acceptor"
     primer_bind     1401..1420
                     /label=Primer 4
     primer_bind     1459..1478
                     /label=T0060_PIRES-F
     primer_bind     complement(1468..1492)
                     /label=#57-R-SA-3R1
     primer_bind     1477..1510
                     /label=B17F-NheI-F
     primer_bind     1477..1492
                     /label=Primer 10
     misc_feature    1492
                     /label=NheI?
     CDS             1493..3561
                     /codon_start=1
                     /gene="YTHDC2"
                     /gene_synonym="CAHL; hYTHDC2"
                     /product="3'-5' RNA helicase YTHDC2 isoform 1"
                     /EC_number="3.6.4.13"
                     /label=YTHDC2
                     /note="isoform 1 is encoded by transcript variant 1;
                     probable ATP-dependent RNA helicase YTHDC2; CsA-associated 
                     helicase-like protein; 3'-5' RNA helicase YTHDC2; YTH 
                     domain containing 2"
                     /db_xref="GI:269847874"
                     /db_xref="CCDS:CCDS4113.1"
                     /db_xref="GeneID:64848"
                     /db_xref="HGNC:HGNC:24721"
                     /db_xref="MIM:616530"
                     /protein_id="NP_073739.3"
                     /translation="RCRPGICFRLFSRLRFQNMLEFQTPELLRMPLQELCLHTKLLAPV
                     NCPIADFLMKAPEPPPALIVRNAVQMLKTIDAMDTWEDLTELGYHLADLPVEPHLGKMV
                     LCAVVLKCLDPILTIACTLAYRDPFVLPTQASQKRAAMLCRKRFTAGAFSDHMALLRAF
                     QAWQKARSDGWERAFCEKNFLSQATMEIIIGMRTQLLGQLRASGFVRARGGGDIRDVNT
                     NSENWAVVKAALVAGMYPNLVHVDRENLVLTGPKEKKVRFHPASVLSQPQYKKIPPANG
                     QAAAIKALPTDWLIYDEMTRAHRIANIRCCSAVTPVTILVFCGPARLASNALQEPSSFR
                     VDGIPNDSSDSEMEDKTTANLAALKLDEWLHFTLEPEAASLLLQLRQKWHSLFLRRMRA
                     PSKPWSQVDEATIRAIIAVLSTEEQSAGLQQPSGIGQRPRPMSSEELPLASSWRSNNSR
                     KSSADTEFSDECTTAERVLMKSPSPALHPPQKYKDRGILHPKRGTEDRSDQSSLKSTDS
                     SSYPSPCASPSPPSSGKGSKSPSPRPNMPVRYFIMKSSNLRNLEISQQKGIWSTTPSNE
                     RKLNRAFWESSIVYLVFSVQGSGHFQGFSRMSSEIGREKSQDWGSAGLGGVFKVEWIRK
                     ESLPFQFAHHLLNPWNDNKKVQISRDGQELEPQVGEQLLQLWERLPLGEKNTTD"
     misc_feature    1493..1494
                     /label=Feature 50
     primer_bind     1680..1700
                     /label=B17F-BsrGI-1F
     primer_bind     2011..2027
                     /label=B17F-SF
     primer_bind     2123..2139
                     /label=B17B-YTHDC2-SF
     misc_feature    3497
                     /label=T
     primer_bind     complement(3540..3582)
                     /label=B17F-BsrGI-R
     primer_bind     complement(3550..3585)
                     /label=B17F-BsrGI-1R
     primer_bind     complement(3550..3567)
                     /label=Primer 1
     misc_feature    3562..3567
                     /label=BsrGI
     CDS             3568..3633
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3573..3673)
                     /label=296
     terminator      3649..4080
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     primer_bind     3649..3674
                     /label=#56-F-W3SL-5F1
     primer_bind     complement(3728..3751)
                     /label=R-Seq-W3SL-5R1
     misc_feature    4123..4263
                     /label=R-ITR
     rep_origin      4266..4721
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4413..4422)
                     /label=F-Seq-Cre-3F1
     promoter        5003..5107
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5108..5968
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="HSNMYPLMRQ*P**MLQ*Y*KRKSMSIQHFRVALIPFFAAFCLPV
                     FAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLS
                     RIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTT
                     IGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLT
                     LASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIV
                     VIYTTG"
     rep_origin      6139..6727
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtcgctc tgcgcgaggc ggctccgccg
      181 cggcggaggg atacggcgca ccatatatat atcgcggggc gcagactcgc gctccggcag
      241 tggtgctggg agtgtcgtgg acgccgtgcc gttactcgta cgccaccatg tccaatttac
      301 tgaccgtaca ccaaaatttg cctgcattac cggtcgatgc aacgagtgat gaggttcgca
      361 agaacctgat ggacatgttc agggatcgcc aggcgttttc tgagcatacc tggaaaatgc
      421 ttctgtccgt ttgccggtcg tgggcggcat ggtgcaagtt gaataaccgg aaatggtttc
      481 ccgcagaacc tgaagatgtt cgcgattatc ttctatatct tcaggcgcgc ggtctggcag
      541 taaaaaccat ccagcaacat ttgggccagc taaacatgct tcatcgtcgg tccgggctgc
      601 cacgaccaag tgacagcaat gctgtttcac tggttatgcg gcggatccga aaagaaaacg
      661 ttgatgccgg tgaacgtgca aaacaggctc tagcgttcga acgcactgat ttcgaccagg
      721 ttcgttcact catggaaaat agcgatcgct gccaggatat acgtaatctg gcatttctgg
      781 ggattgctta taacaccctg ttacgtatag ccgaaattgc caggatcagg gttaaagata
      841 tctcacgtac tgacggtggg agaatgttaa tccatattgg cagaacgaaa acgctggtta
      901 gcaccgcagg tgtagagaag gcacttagcc tgggggtaac taaactggtc gagcgatgga
      961 tttccgtctc tggtgtagct gatgatccga ataactacct gttttgccgg gtcagaaaaa
     1021 atggtgttgc cgcgccatct gccaccagcc agctatcaac tcgcgccctg gaagggattt
     1081 ttgaagcaac tcatcgattg atttacggcg ctaaggatga ctctggtcag agatacctgg
     1141 cctggtctgg acacagtgcc cgtgtcggag ccgcgcgaga tatggcccgc gctggagttt
     1201 caataccgga gatcatgcaa gctggtggct ggaccaatgt aaatattgta atgaactata
     1261 tccgtaacct ggatagtgaa acaggggcaa tggtgcgcct gctggaagat ggcgattagg
     1321 taagtatcaa ggttacaaga caggtttaag gagaccaata actagtataa cttcgtatag
     1381 gatactttat agcaatttat gaaactgggc ttgtcgagac agagaagact cttgcgtttc
     1441 tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac agggcgatgt
     1501 agacctggaa tttgttttcg tctgttcagt agactccgat tccagaatat gttggaattt
     1561 cagactccgg aacttttgag aatgccatta caggaacttt gcttacatac caagctgtta
     1621 gccccagtta attgtcccat tgctgatttt cttatgaaag ctcctgaacc tccaccagct
     1681 ttaattgtaa gaaatgctgt acaaatgctt aagacaatag atgcaatgga tacatgggaa
     1741 gatctgactg aacttgggta tcatttggct gacttgccag tagaaccaca tcttggtaaa
     1801 atggtcttgt gtgctgttgt tttaaagtgt ctggacccca tccttacaat tgcttgcaca
     1861 ctagcttatc gagatccttt tgtactacct actcaggcct ctcaaaaacg tgcagctatg
     1921 ctttgtagga aacgttttac tgcaggagct ttcagtgacc atatggcact tctcagagca
     1981 ttccaggcct ggcaaaaagc acgaagtgat gggtgggagc gagccttttg tgaaaagaat
     2041 tttctttcac aggctactat ggaaataatc ataggcatga gaacacagtt gcttggtcaa
     2101 cttagagcat caggttttgt tagagcacga ggtggtggtg acattcggga cgttaacaca
     2161 aactctgaga attgggctgt cgttaaagct gcattggtgg caggcatgta tcctaattta
     2221 gtccacgtgg acagagagaa tctagtgttg acagggccaa aggagaaaaa agtacgattt
     2281 catcctgctt cagttctcag tcagcctcaa tataaaaaga ttcctccagc caatggtcaa
     2341 gctgcagcaa ttaaggcact gcccacagat tggcttattt atgatgaaat gaccagagcc
     2401 catagaatag ctaatattag atgttgttca gcagtgacgc ctgtcactat attggtattc
     2461 tgtggaccag ctagattggc aagtaatgct cttcaggaac cttcatcctt tagagtggat
     2521 ggcattccca atgacagtag tgatagtgaa atggaggaca aaactacagc taatttggca
     2581 gccttgaaac ttgatgagtg gctccatttc acactggagc cagaggcagc tagtttattg
     2641 ctgcagctca gacagaagtg gcatagctta tttttacgcc gaatgagagc tccatctaaa
     2701 ccttggtctc aagttgatga agctaccata agagcaatta tagctgtttt aagcactgaa
     2761 gaacagtctg caggtttaca acaaccatct gggattggcc aaaggccaag gcctatgtct
     2821 tcagaagagc ttcctttggc ctcatcttgg aggtcaaata atagtaggaa aagttcagca
     2881 gatactgaat tttctgatga gtgtactact gcagaaagag tactgatgaa atctccatct
     2941 ccagcattac acccacctca gaagtacaaa gatagaggaa ttttacatcc taaacgaggt
     3001 actgaggacc gatcagatca gtcttctctg aaatctacag acagcagtag ttacccaagt
     3061 ccttgtgcta gtccttctcc tccatcctca ggaaagggct caaaatctcc ttcgccaaga
     3121 ccaaacatgc ctgttcgata cttcataatg aagagtagca atttgagaaa ccttgaaatt
     3181 tctcaacaga agggtatctg gtctacaact cctagtaatg aacggaagct aaatcgagcc
     3241 ttttgggaaa gcagcatagt ttacttggta ttttctgttc aaggatctgg acatttccag
     3301 ggattttcta ggatgtcttc tgagattgga agggaaaaga gtcaggactg gggctctgct
     3361 ggactaggag gagtatttaa ggtggagtgg atacgaaaag aaagccttcc ctttcaattt
     3421 gcacaccatt tactcaatcc atggaatgac aacaagaaag tgcagataag cagggatggg
     3481 caggaactag aacctcaggt tggtgaacag ttgctccagt tatgggaacg tcttcccttg
     3541 ggagaaaaaa acacaactga ttgtacagac tacaaagacc atgacggtga ttataaagat
     3601 catgacatcg actacaagga tgacgatgac aagtaagaat tctgatgact cgagataatc
     3661 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     3721 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     3781 ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg gaactcatcg
     3841 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     3901 tgtttatttg tgaaatttgt gatgctattg ctttatttgt aaccatctag ctttatttgt
     3961 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     4021 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa
     4081 agatctggag accggtgtgg cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     4141 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     4201 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     4261 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     4321 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4381 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     4441 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4501 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     4561 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4621 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     4681 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     4741 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     4801 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     4861 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     4921 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     4981 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     5041 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5101 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5161 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5221 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5281 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5341 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5401 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5461 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     5521 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5581 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5641 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5701 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5761 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5821 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5881 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5941 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6001 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6061 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6121 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6181 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6241 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6301 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6361 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6421 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6481 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6541 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6601 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6661 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6721 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6781 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6841 ctggcctttt gctcacatgt
//
