LOCUS       sequence-326292-        4429 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 4429)
  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 4429)
  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..4429
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    37..261
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(982..1015)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      complement(1041..1088)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1084..1102
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             2011..2019
                     /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)"
     protein_bind    complement(2063..2096)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(2152..2607)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2289..2310)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2501..2520
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2721..3536)
                     /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             complement(2721..3536)
                     /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     3444..3463
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3537..3639)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      3754..4342
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4243..4262
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtggataacc gtgtcagaat tcgtctcatg acagcgccat agagcccacc gcatccccag
       61 catgcctgct attgtcttcc caatcctccc ccttgctgtc ctgccccacc ccacccccca
      121 gaatagaatg acacctactc agacaatgcg atgcaatttc ctcattttat taggaaagga
      181 cagtgggagt ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa acaacagatg
      241 gctggcaact agaaggcaca gaagcttact tgtacagctc atccatgccg cccgtgctat
      301 gacgtccttc ggcacgctcg tactgctcca cgatggtgta gtcctcgttg tggctggtga
      361 tgtccagctt gatgttcacg ttgtaggcgc cgggcaattg cacgggcttc ttggccttgt
      421 aggtggtctt cacctcggcg tcgtagtggc ctccatcctt cagcttcagg cgctgcttga
      481 tctcgccctt cagagcgccg tcctcgggat acatgcgctc gctggaagcc tcccagccca
      541 tggtcttctt ctgcatcacc gggccatcgg agggaaagtt cgtgccgcgc agcttgacct
      601 tgtagatgaa ctcgccgtcc tgcaagctgg agtcctgggt cacggtcacc actccgccgt
      661 cctcgaagtt catcacgcgc tcccacttga agccttcggg gaagctcagc ttcaggtagt
      721 cgggaatgtc ggccgggtgc ttcacgtagg ccttggagcc gtacatgaac tgggggctca
      781 agatgtccca ggcaaagggc aacgggcctc ccttggtcac cttcagtttg gcggtctggg
      841 tgccttcgta gggacgtccc tcgccttcgc cctcgatctc gaactcgtgg ccattcacgc
      901 tgccctccat gtggaccttg aagcgcatga actccttgat gatggccatg ttatcctctt
      961 ctcccttgga caccatcgag gataacttcg tatagcatac attatacgaa gttatatccg
     1021 gatatagttc ctcctttcag caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
     1081 tatgctagtt attgctcagc ggtggcagca gccaactcag cttcctttcg ggctttgaag
     1141 cttaccaatg cttaataaga gaagccccga tttctgcgat ctgacggtta cgctcgtcca
     1201 tcgtcgcctg ggagcctgtc gtatagatta ccacaatacg tgagggtttc ccatccggac
     1261 caagtgctgc aataatgcca cgagaaccgc gttctcccgc cccagactta tccgcaataa
     1321 accatccggc tggcagggca gagcgcagca gcggcccggc cactttatct gcctccatcc
     1381 aatcgataag ttgctggcga gatgccaggg ttaaaagctc tccagtcagc aatttacgca
     1441 gcgttgtggc catagcgacg ggcatcgtcg tgtcacgttc gtcattgggg atagcttcgt
     1501 ttaattccgg ctcccagcgg tctaaacgag tgacgtgatc gcccatgttg tgaaggaatg
     1561 ccgtcagctc cttcggaccg ccaatggtcg taagcagaag gttggcggcg gtgttatcag
     1621 acatcgtaat agcggcacta cacagttcac gaactgtcat gccatcagtc aaatgcttct
     1681 ctgttacggg ggagtattcc acaagatcat tctgggagta gtggatacga cggcccagtt
     1741 gctcttgacc tgcatcgatg gagcttaaaa ccgcgccgca caacaacact ttgaaggtac
     1801 tcatcatggg gaaacgttct tcagggcgga agctctctaa gatttttccg gaattaaggt
     1861 ccaactcaat gtaaccgaca cgagcaccaa gttgatcctc tgcatcctta actttgacca
     1921 gcgtttctgg atgggcgaat actggcaaac agaatgcagc aaaaaagggg atcagcgcca
     1981 cacggaaatg ttgaatactc attatgtcta acctccttac gtatcgcata ttatatgtga
     2041 atcacagtga tatgtaagta ttataacttc gtatagcata cattatacga agttatggca
     2101 ctgccgccgc tacctccgct tccaccggaa cctcctgaga cgaattcgtt aaaattgtaa
     2161 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     2221 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     2281 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     2341 ggcgaaaaac cgtctatcag ggcgctggcc cactacgtga accatcaccc taatcaagtt
     2401 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     2461 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     2521 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     2581 cgcttaatgc gccgctacag ggcgcgtaac atcatgtttg acagcttatc atcgaatttc
     2641 tgccattcat ccgcttatta tcacttattc aggcgtagca accaggcgtt taagggcacc
     2701 aataactgcc ttaaaaaaaa tcagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
     2761 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     2821 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     2881 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     2941 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc
     3001 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     3061 ccgttattca ttcgtgattg cgcctgagcg agtcgaaata cgcgatcgct gttaaaagga
     3121 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     3181 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc tgggatcgca
     3241 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     3301 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     3361 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcggtagatt
     3421 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     3481 atgttggaat ttaatcgcgg ccttgagcaa gacgtttccc gttgaatatg gctcatttta
     3541 gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt
     3601 tcattatggt gaaagttgga acctcttacg tgccgatcat cacaaaagga tctaggtgaa
     3661 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3721 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3781 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3841 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3901 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3961 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4021 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4081 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4141 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4201 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4261 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4321 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4381 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattct
//
