LOCUS       sequence-349454-        6074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry mRNA from CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6074)
  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 6074)
  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: "Nucleic 
            Acids Res. 2022 Mar 4. pii: 6542487. doi: 10.1093/nar/gkac159."
COMMENT     Sequence Label: pcDNA3.1(+) mCherry
FEATURES             Location/Qualifiers
     source          1..6074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(26..45)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        217..596
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        597..800
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     751..771
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     845..864
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        845..863
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      899..908
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             905..1615
                     /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(1050..1068)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1318..1337
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1647..1664)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1653..1877
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1923..2351
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2010..2029)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2220..2241
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2360..2380)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2365..2694
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2545..2680
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2607..2626
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2761..3555
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2815..2834)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3425..3444
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3729..3850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3766..3785)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3820..3839
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3899..3915)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3899..3915)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3912..3934)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3923..3939
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3947..3953,3954..3971,3972..3977))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3992..4013
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4130..4147)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4301..4889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4381..4400)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5060..5851,5852..5920))
                     /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     5683..5702
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5921..6025)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
       61 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
      121 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
      181 cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt
      241 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      301 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      361 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      421 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      481 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      541 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      601 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      661 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      721 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      781 aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg
      841 aaattaatac gactcactat agggagaccc aagctggcta gcgtttaaac ttaagcttgc
      901 caccatggtg agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt
      961 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
     1021 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
     1081 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
     1141 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
     1201 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
     1261 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
     1321 cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga
     1381 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
     1441 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
     1501 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     1561 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca agtgatctag
     1621 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     1681 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     1741 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     1801 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     1861 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     1921 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     1981 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2041 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2101 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2161 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2221 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2281 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2341 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     2401 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     2461 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2521 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     2581 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     2641 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     2701 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     2761 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     2821 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     2881 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     2941 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3001 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3061 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3121 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     3181 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     3241 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     3301 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     3361 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     3421 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     3481 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     3541 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     3601 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     3661 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     3721 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3781 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3841 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     3901 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     3961 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4021 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4081 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4141 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4201 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4261 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4321 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4381 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4441 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4501 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4561 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4621 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4681 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4741 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4801 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4861 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4921 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4981 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5041 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5101 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5161 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5221 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5281 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5341 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5401 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5461 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5521 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5581 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5641 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5701 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5761 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5821 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5881 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5941 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6001 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     6061 gatcgggaga tctc
//
