LOCUS       sequence-346360-        9442 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses mCherry mRNA from the Drosophila Ubi-p63e promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9442)
  AUTHORS   Ai Y, Liang D, Wilusz JE
  TITLE     CRISPR/Cas13 effectors have differing extents of off-target effects 
            that limit their utility in eukaryotic cells.
  JOURNAL   Nucleic Acids Res. 2022 Mar 4. pii: 6542487. doi: 
            10.1093/nar/gkac159.
  PUBMED    35244715
REFERENCE   2  (bases 1 to 9442)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 2022 Mar 4. pii: 6542487. doi: 10.1093/nar/gkac159."
COMMENT     Sequence Label: Hy_pUbi-p63e mCherry SV40
FEATURES             Location/Qualifiers
     source          1..9442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          14..79
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             209..229
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    654..775
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             2816..3526
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2961..2979)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3229..3248
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3535..3656
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4366..4383)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4537..5125)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4617..4636)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5296..6087,6088..6156))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5919..5938
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6157..6261)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6329..6347
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7743..8022
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     CDS             8068..9093
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             8068..9093
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
ORIGIN
        1 ttaaagctct aaggtaaata taaaattttt aagtgtataa tgtgttaaac tactgattct
       61 aattgtttgt gtattttaga ttccaaccta tggaactgat gaatgggagc agtggtggaa
      121 tgcctttaat gaggaaaacc tgttttgctc agaagaaatg ccatctagtg atgatgaggc
      181 tactgctgac tctcaacatt ctactcctcc aaaaaagaag agaaaggtag aagaccccaa
      241 ggactttcct tcagaattgc taagtttttt gagtcatgct gtgtttagta atagaactct
      301 tgcttgcttt gctatttaca ccacaaagga aaaagctgca ctgctataca agaaaattat
      361 ggaaaaatat tctgtaacct ttataagtag gcataacagt tataatcata acatactgtt
      421 ttttcttact ccacacaggc atagagtgtc tgctattaat aactatgctc aaaaattgtg
      481 tacctttagc tttttaattt gtaaaggggt taataaggaa tatttgatgt atagtgcctt
      541 gactagagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
      601 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
      661 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
      721 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
      781 tctggattaa ttaacttgtc gccggaacgc agcgacagag attccaatgt gtccgtatct
      841 ttcaggcttt tgcccttcag ttccagacga agcgactggc gattcgcgtg tggggtctgc
      901 ttcagggtct tgtgaattag ggcgcgcaga tcgccgatgg gcgtggcgcc ggagggcacc
      961 ttcaccttgc cgtacggctt gctgttcttc gcgttcaaaa tctccagctc cattttgctt
     1021 tcggtgcgct tgcaatcagt actgtccaaa atcgaaaatc gccgaaccgt agtgtgaccg
     1081 tgcggggctc tgcgaaaata aactttttta ggtatatggc cacacacggg gaaagcacag
     1141 tggattatat gttttaatat tataatatgc aggttttcat tacttatcca gatgtaagcc
     1201 cacttaaagc gatttaacaa ttatttgccg aaagagtata aacaaatttc acttaaaaat
     1261 ggattaagaa aagctagctt gtgtaagatt atgcgcagcg ttgccagata gctccattta
     1321 aaacacttca aaaacaataa gttttgaaaa tatatacata aatagcagtc gttgccgcaa
     1381 cgctcaacac atcacacttt taaaacaccc tttacctaca cagaattact ttttaaattt
     1441 ccagtcaagc tgcgagtttc aaaattatag ccggtagaga agacagtgct atttcaaaag
     1501 caaactaaca agggtcttaa attccaaaac accaatccta acaagccttg gacttttgta
     1561 agtttagatc aaaggtggca ttgcattcaa tgtcatggta agaagtaggt cgtctaggta
     1621 gaaatcctca ttcagccggt caagtcagta cgagaaaggt ctcaatttga aattgtctta
     1681 aaaatatttt attgttttgt actgtggtga gtttaaacga aaaacacaaa aaaaaagtga
     1741 tacacagaaa tcataaaaaa ttttaataca aggtattcgt acgtatcaaa aacatttcgg
     1801 cacaattttt tttctctgta ctaaagtgtt acgaacacta cggtattttt tagtgatttt
     1861 caacggacac cgaaggtata taaacagcgt tcgcgaacgg tcgccttcaa aaccaattga
     1921 catttgcagc agcaagtaca agtagaaagt aaagcgcaat cagcgaaaaa tttatactta
     1981 attgttggtg attaaagtac aattaaaaga acattctcga aagtcacaag aaacgtaagt
     2041 ttttaactcg ctgttaccaa ttagtaataa gagcaacaag acgttgagta atttcaagaa
     2101 aaactgcatt tcaaggtctt tgttcggcca ttttttttta ttcaacgctc tacgtaatta
     2161 caaaataaga aattggcagc cacgcatctt gttttcccaa tgaattggca tcaaaacgca
     2221 aacaaatcta taaataaaac ttgcgtgttg attttcgcca agatttattg gcaaattgtg
     2281 aaattcgcag tgacgcattt gaaaattcga gaaatcacga acgcactcga tcgagcattt
     2341 gtgtgcatgt tattagttag ttgtttagtt aattgaagta ttttaccaac gaaatccact
     2401 tatttttagc tgaaatagag taggttgctt aaacaaagcc acgtctgaaa atttcttatt
     2461 gcttgtagtt gtgacgtcac catatacaca caaaataatg tgtatgcatg catttcagct
     2521 gtgtatatat acatgcacac actcgcaaca cgaaaacgat gacgaagcaa cggaacaaag
     2581 gtttctcaac taccctttgt tccctgtttc ttcgctttcc tttgttccaa tattcgtaga
     2641 gggttaatag gggtttctca acaaagttgg cgtcgataaa taagtttccc atttttattc
     2701 cccagccagg aagttagttt caatagtttt gtaatttcaa cgaaactcat ttgatttcgt
     2761 actaattttc cacatctcta ttttcttccc gcagaataat ccatctagag ccaaaatggt
     2821 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
     2881 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     2941 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     3001 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
     3061 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     3121 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     3181 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     3241 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     3301 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     3361 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     3421 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     3481 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtgagcgg ccgcaacttg
     3541 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3601 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacta
     3661 gtggatccac taggggccgc cgacgcgagg ctggatggcc ttccccatta tgattcttct
     3721 cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga
     3781 cgaccatcag ggacagcttc aaggatcgct cgcggctctt accagcctaa cttcgatcat
     3841 tggaccgctg atcgtcacgg cgatttatgc cgcctcggcg agcacatgga acgggttggc
     3901 atggattgta ggcgccgccc tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg
     3961 gagccgggcc acctcgacct gaatggaagc cggcggcacc tcgctaacgg attcaccact
     4021 ccaagaattg gagccaatca attcttgcgg agaactgtga atgcgcaaac caacccttgg
     4081 cagaacatat ccatcgcgtc cgccatctcc agcagccgca cgcggcgcat ctcgggcagc
     4141 gttgggtcct ggccacgggt gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc
     4201 tggcggggtt gccttactgg ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc
     4261 gactgctgct gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt cggtttccgt
     4321 gtttcgtaaa gtctggaaac gcggaagtca gcgctcttcc gcttcctcgc tcactgactc
     4381 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4441 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4501 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4561 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4621 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4681 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4741 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4801 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4861 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4921 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     4981 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5041 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5101 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5161 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5221 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5281 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5341 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5401 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5461 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5521 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5581 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
     5641 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5701 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5761 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5821 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5881 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     5941 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6001 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6061 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6121 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6181 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6241 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     6301 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
     6361 agaattcaca tttgtacgaa tttttttttt atcaaaagtt cgagtttttc accaatttcc
     6421 tcatcaaccg agcaaggcaa acggctttga ataatatggt gttatatata catatatcaa
     6481 atcgctgctg actgcgtgat tgatggcccc aagattacat attatcgaat caggattcag
     6541 aaggagatca atgtcaaatg cggacaggaa catgaaagac gcctgttatg cgcaattaaa
     6601 aatttgggtt taattgctgt ggaaactgtt gttggcggca tcttaagttc ctgtttaaca
     6661 acatcaacta cttatgtacg tagaagcgtt taagccattt gcatacagat gagaactggc
     6721 ttttgtgcta atcagtcaag atgactccga tgatgatgac tcattacctg accagttttc
     6781 gctgctttct tttcaacaac tacttgtata tgtattgtat ccaatagcaa tacattgaat
     6841 ttccatggtc tagtcacgta ttatcattta attgacacca agtcgtgtta ttgttgagct
     6901 atcgagttca gctcaaacat ttcttattcc catgaataag ccggcaaaaa tatgcaatct
     6961 atgaaagtta atataagcaa accttacttt gactcaatac caatgcactt tgtgtcgata
     7021 ggttcacgca attgaggcga ttattccgat aacccaagcg attgactgtt cccgtttcga
     7081 ttccaattga aatttggaaa tgtacaatag ttttgctata tgctgtcaag tacgctctta
     7141 tcttctctgg gttttcttca gagtttcgaa acgcttcttc ttttttttgt tttttttttt
     7201 ttggaatctc gtattttgga aggggctccc ctctggaatt tgttacactg tcgttatcat
     7261 tgcgaacaag cggcccgaag ctatcagcga ctttaacatt tacaatgcac ttttttacga
     7321 ccaattaaat gtacattttc ctttcttcgc ccgttgataa gcgaacgcga tgtggcgcag
     7381 gcaatgtgtt gctcttgcga cacaaacgca atcaaaatgg attcaatttc gctttttccc
     7441 agtgaaacga agaacgaacc gaccatcatg atatgctcct ctgcatgttg cgtattgaat
     7501 caatgacaat ttcaattaag ccgcccgttc gtcatgcgtt ttcgtgcgct tcgaaatgct
     7561 gataacgctg ctgtcctcca actgctttgc atgtggacac aattccattt atttaattct
     7621 tttatttgga tcggttaaat taaaaagcgc cttgttacgc atttaacgtt gtttccggtg
     7681 cgtggtggtt tcatgcttct gggaacggca aatgggttta ggattgggaa cccctcatca
     7741 tctgttggaa tatactattc aacctacaaa agtaacgtta aacaacacta ctttatattt
     7801 gatatgaatg gccacacctt ttatgccata aaacatattg taagagaata ccactctttt
     7861 tattccttct ttccttcttg tacgtttttt gctgtaagta ggtcgtggtg ctggtgttgc
     7921 agttgaaata acttaaaata taaatcataa aactcaaaca taaacttgac tatttattta
     7981 tttattaaga aaggaaatat aaattataaa ttacaacagg ttatggacct gcagccaagc
     8041 ttggcgctcg tccgggggca atgagatatg aaaaagcctg aactcaccgc gacgtctgtc
     8101 gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc
     8161 gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat
     8221 agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg
     8281 ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc
     8341 tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt
     8401 ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc
     8461 gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata
     8521 tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt
     8581 gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc
     8641 cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata
     8701 acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac
     8761 atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg
     8821 aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt
     8881 gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt
     8941 cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc
     9001 agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga
     9061 cgccccagca ctcgtccgag ggcaaaggaa tagagtagat gccgaccgaa caagagctga
     9121 tttcgagaac gcctcagcca gcaactcgcg cgagcctagc aaggcaaatg cgagagaacg
     9181 gccttacgct tggtggcaca gttctcgtcc acagttcgct aagctcgctc ggctgggtcg
     9241 cgggagggcc ggtcgcagtg attcaggccc ttctggattg tgttggtccc cagggcacga
     9301 ttgtcatgcc cacgcactcg ggtgatctga ctgatcccgc agattggaga tcgccgcccg
     9361 tgcctgccga ttgggtgcag atctttgtga aggaacctta cttctgtggt gtgacataat
     9421 tggacaaact acctacagag at
//
