LOCUS       sequence-328915-        5916 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry2 under the control of CMV promoter, basic vector 
            for AAV, pAAV-CMV backbone, multiclonig site at C terminal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5916)
  AUTHORS   Tanida I, Furuta Y, Yamaguchi J, Kakuta S, Oliva Trejo JA, Uchiyama 
            Y
  TITLE     Two-color in-resin CLEM of Epon-embedded cells using osmium 
            resistant green and red fluorescent proteins.
  JOURNAL   Sci Rep. 2020 Dec 14;10(1):21871. doi: 10.1038/s41598-020-78879-x.
  PUBMED    33318540
REFERENCE   2  (bases 1 to 5916)
  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: "Sci Rep."; 
            date: "2020-12-14"; volume: "10(1):21871. doi"; pages: " 
            10.1038/s41598-020-78879-x"
COMMENT     Sequence Label: pAC-mCherry2-G4
FEATURES             Location/Qualifiers
     source          1..5916
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    157..178
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(193..198,199..216,217..223)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    231..247
                     /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)."
     primer_bind     236..258
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     255..271
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     255..271
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   287..427
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   298..427
                     /label=AAV2 ITR
     enhancer        496..799
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        800..1003
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     954..974
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          1094..1472
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     regulatory      1579..1588
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1585..2292
                     /codon_start=1
                     /product="derivative of the monomeric red fluorescent 
                     protein mCherry"
                     /label=mCherry2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFNWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQCRTMGWEASTERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVDIKLDILSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1730..1748)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    2350..2406
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2456..2932
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2655..2674)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2945..3085
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2945..3074
                     /label=AAV2 ITR
     primer_bind     complement(3095..3112)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3095..3111)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3104..3126)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3324..3779
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3411..3430)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3621..3642
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3796..3815)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3915..3937
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3975..3993)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4061..4165
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4166..4234,4235..5026)
                     /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(4384..4403)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5197..5785
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5686..5705
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
       61 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      121 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      181 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      241 ataacaattt cacacaggaa acagctatga ccatgattac gaattgcctg caggcagctg
      301 cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt
      361 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
      421 ggttcctatc gatatcaagc tttaatagta atcaattacg gggtcattag ttcatagccc
      481 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      541 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      601 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      661 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      721 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      781 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      841 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      901 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      961 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tggatatcct
     1021 taagggccca gccggcccga atcccggccg ggaacggtgc attggaacgc ggattccccg
     1081 tgccaagagt gacgtaagta ccgcctatag agtctatagg cccacaaaaa atgctttctt
     1141 cttttaatat acttttttgt ttatcttatt tctaatactt tccctaatct ctttctttca
     1201 gggcaataat gatacaatgt atcatgcctc tttgcaccat tctaaagaat aacagtgata
     1261 atttctgggt taaggcaata gcaatatttc tgcatataaa tatttctgca tataaattgt
     1321 aactgatgta agaggtttca tattgctaat agcagctaca atccagctac cattctgctt
     1381 ttattttatg gttgggataa ggctggatta ttctgagtcc aagctaggcc cttttgctaa
     1441 tcatgttcat acctcttatc ttcctcccac agctcctggg caacgtgctg gtctgtgtgc
     1501 tggcccatca ctttggcaaa gaattgggat tcgcgaattg tgctgtctca tcattttggc
     1561 aaagctagcc taccggtcgc caccatggtg agcaagggcg aggaggataa catggccatc
     1621 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
     1681 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     1741 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
     1801 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
     1861 cccgagggct tcaattggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     1921 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     1981 aacttcccct ccgacggccc cgtaatgcag tgtcgtacca tgggctggga ggcctccact
     2041 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     2101 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     2161 cagctgcccg gcgcctacaa cgtcgacatc aagttggaca tcctttccca caacgaggac
     2221 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     2281 gagctgtaca agtccggact caccggtgga ggcggttcag gcggaggtgg ctctggcggt
     2341 ggcggatcga gatctcgagc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg
     2401 ggatccaccg gatctagata actgatcaac tcctcaggtg caggctgcct atcacgggtg
     2461 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
     2521 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
     2581 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
     2641 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
     2701 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg
     2761 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt
     2821 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc
     2881 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttatcgatag
     2941 atctaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     3001 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     3061 gagcgagcgc gcagctgcct gcaggcagct tggcactggc cgtcgtttta caacgtcgtg
     3121 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     3181 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     3241 atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3301 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3361 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3421 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3481 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     3541 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3601 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3661 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3721 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     3781 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3841 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3901 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3961 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4021 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4081 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4141 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4201 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4261 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4321 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4381 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4441 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4501 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4561 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4621 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4681 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4741 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4801 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4861 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4921 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4981 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5041 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5101 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5161 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5221 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5281 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5341 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5401 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5461 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5521 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5581 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5641 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5701 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5761 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5821 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     5881 ccgtattacc gcctttgagt gagctgatac cgctcg
//
