LOCUS       sequence-323386-        4400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for amplification of mcherry C-terminal tagging cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4400)
  AUTHORS   Matinyan N, Karkhanis MS, Gonzalez Y, Jain A, Saltzman A, 
            Malovannaya A, Sarrion-Perdigones A, Dierick HA, Venken KJT
  TITLE     Multiplexed drug-based selection and counterselection genetic 
            manipulations in Drosophila.
  JOURNAL   Cell Rep. 2021 Sep 14;36(11):109700. doi: 
            10.1016/j.celrep.2021.109700.
  PUBMED    34525356
REFERENCE   2  (bases 1 to 4400)
  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: "Cell Rep.";
            date: "2021-09-14"; volume: "36(11):109700. doi"; pages: " 
            10.1016/j.celrep.2021.109700"
COMMENT     Sequence Label: pmCherry-CT-Tagging
FEATURES             Location/Qualifiers
     source          1..4400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    56..89
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     RBS             134..142
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(1051..1069)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1065..1112
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     protein_bind    1138..1171
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             join(1177..1179,1180..1182,1183..1896)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1177..1179,1180..1182,1183..1893)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1222..1243)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1483..1502)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1830..1851
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1901..2125
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2249..2837)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2329..2348)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2952..3054
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3055..3870
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             3055..3870
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3128..3147)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4010..4390
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4097..4116)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4307..4328
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 taacgaattc gtctcaggag gttccggtgg aagcggaggt agcggcggca gtgccataac
       61 ttcgtataat gtatgctata cgaagttata atacttgaca tatcactgtg attcacatat
      121 aatatgcgat acgtaaggag gttagacata atgagtattc aacatttccg tgtggcgctg
      181 atcccctttt ttgctgcatt ctgtttgcca gtattcgccc atccagaaac gctggtcaaa
      241 gttaaggatg cagaggatca acttggtgct cgtgtcggtt acattgagtt ggaccttaat
      301 tccggaaaaa tcttagagag cttccgccct gaagaacgtt tccccatgat gagtaccttc
      361 aaagtgttgt tgtgcggcgc ggttttaagc tccatcgatg caggtcaaga gcaactgggc
      421 cgtcgtatcc actactccca gaatgatctt gtggaatact cccccgtaac agagaagcat
      481 ttgactgatg gcatgacagt tcgtgaactg tgtagtgccg ctattacgat gtctgataac
      541 accgccgcca accttctgct tacgaccatt ggcggtccga aggagctgac ggcattcctt
      601 cacaacatgg gcgatcacgt cactcgttta gaccgctggg agccggaatt aaacgaagct
      661 atccccaatg acgaacgtga cacgacgatg cccgtcgcta tggccacaac gctgcgtaaa
      721 ttgctgactg gagagctttt aaccctggca tctcgccagc aacttatcga ttggatggag
      781 gcagataaag tggccgggcc gctgctgcgc tctgccctgc cagccggatg gtttattgcg
      841 gataagtctg gggcgggaga acgcggttct cgtggcatta ttgcagcact tggtccggat
      901 gggaaaccct cacgtattgt ggtaatctat acgacaggct cccaggcgac gatggacgag
      961 cgtaaccgtc agatcgcaga aatcggggct tctcttatta agcattggta agcttcaaag
     1021 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     1081 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatata
     1141 acttcgtata atgtatgcta tacgaagtta tcctcgatgg tgagcaaggg cgaggagctg
     1201 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1261 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     1321 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     1381 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1441 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1501 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1561 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1621 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     1681 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     1741 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     1801 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1861 gggatcactc tcggcatgga cgagctatac aagtaagctt ctgtgccttc tagttgccag
     1921 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     1981 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     2041 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     2101 gctggggatg cggtgggctc tatggcgctg tcatgagacg aattctgaca cggttatcca
     2161 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2221 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2281 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2341 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2401 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2461 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2521 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2581 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2641 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     2701 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2761 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2821 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2881 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2941 tccttttgtg atgatcggca cgtaagaggt tccaactttc accataatga aataagatca
     3001 ctaccgggcg tattttttga gttatcgaga ttttcaggag ctaaggaagc taaaatgagc
     3061 catattcaac gggaaacgtc ttgctcaagg ccgcgattaa attccaacat ggatgctgat
     3121 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctaccga
     3181 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     3241 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     3301 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatccca
     3361 gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     3421 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     3481 agcgatcgcg tatttcgact cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     3541 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     3601 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     3661 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     3721 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     3781 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     3841 tttcatttga tgctcgatga gtttttctga ttttttttaa ggcagttatt ggtgccctta
     3901 aacgcctggt tgctacgcct gaataagtga taataagcgg atgaatggca gaaattcgat
     3961 gataagctgt caaacatgat gttcacattt ccccgaaaag tgccacctga cgcgccctgt
     4021 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4081 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4141 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4201 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4261 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4321 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4381 ccgatttcgg cctattggtt
//
