LOCUS       sequence-328278-        4420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for amplification of mCherry N-terminal tagging cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4420)
  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 4420)
  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-NT-Tagging
FEATURES             Location/Qualifiers
     source          1..4420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(97..685)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(177..196)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        800..902
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             903..1718
                     /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             903..1718
                     /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(976..995)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1832..2287
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1919..1938)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2129..2150
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2311..2522
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     protein_bind    3238..3271
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     RBS             3316..3324
                     /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(4233..4251)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4247..4294
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     protein_bind    4320..4353
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
       61 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      121 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      181 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      241 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      301 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      361 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      421 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
      481 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
      541 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
      601 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
      661 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
      721 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
      781 cacctagatc cttttgtgat gatcggcacg taagaggttc caactttcac cataatgaaa
      841 taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta
      901 aaatgagcca tattcaacgg gaaacgtctt gctcaaggcc gcgattaaat tccaacatgg
      961 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     1021 tctaccgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     1081 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     1141 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     1201 cgatcccagg gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     1261 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     1321 cttttaacag cgatcgcgta tttcgactcg ctcaggcgca atcacgaatg aataacggtt
     1381 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     1441 aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     1501 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     1561 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     1621 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     1681 aattgcagtt tcatttgatg ctcgatgagt ttttctgatt ttttttaagg cagttattgg
     1741 tgcccttaaa cgcctggttg ctacgcctga ataagtgata ataagcggat gaatggcaga
     1801 aattcgatga taagctgtca aacatgatgt tacgcgccct gtagcggcgc attaagcgcg
     1861 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     1921 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     1981 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2041 cttgattagg gtgatggttc acgtagtggg ccagcgccct gatagacggt ttttcgccct
     2101 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     2161 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     2221 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     2281 acaattttaa cgaattcgtc tcaggagtat gggcagagcg cacatcgccc acagtccccg
     2341 agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa
     2401 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
     2461 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
     2521 agaatggtgt ccaagggaga agaggataac atggccatca tcaaggagtt catgcgcttc
     2581 aaggtccaca tggagggcag cgtgaatggc cacgagttcg agatcgaggg cgaaggcgag
     2641 ggacgtccct acgaaggcac ccagaccgcc aaactgaagg tgaccaaggg aggcccgttg
     2701 ccctttgcct gggacatctt gagcccccag ttcatgtacg gctccaaggc ctacgtgaag
     2761 cacccggccg acattcccga ctacctgaag ctgagcttcc ccgaaggctt caagtgggag
     2821 cgcgtgatga acttcgagga cggcggagtg gtgaccgtga cccaggactc cagcttgcag
     2881 gacggcgagt tcatctacaa ggtcaagctg cgcggcacga actttccctc cgatggcccg
     2941 gtgatgcaga agaagaccat gggctgggag gcttccagcg agcgcatgta tcccgaggac
     3001 ggcgctctga agggcgagat caagcagcgc ctgaagctga aggatggagg ccactacgac
     3061 gccgaggtga agaccaccta caaggccaag aagcccgtgc aattgcccgg cgcctacaac
     3121 gtgaacatca agctggacat caccagccac aacgaggact acaccatcgt ggagcagtac
     3181 gagcgtgccg aaggacgtca tagcacgggc ggcatggatg agctgtacaa gagtgccata
     3241 acttcgtata atgtatgcta tacgaagtta taatacttga catatcactg tgattcacat
     3301 ataatatgcg atacgtaagg aggtcagaca taatgagtat tcaacatttc cgtgtggcgc
     3361 tgatcccctt ttttgctgca ttctgtttgc cagtattcgc ccatccagaa acgctggtca
     3421 aagttaagga tgcagaggat caacttggtg ctcgtgtcgg ttacattgag ttggacctta
     3481 attccggaaa aatcttagag agcttccgcc ctgaagaacg tttccccatg atgagtacct
     3541 tcaaagtgtt gttgtgcggc gcggttttaa gctccatcga tgcaggtcaa gagcaactgg
     3601 gccgtcgtat ccactactcc cagaatgatc ttgtggaata ctcccccgta acagagaagc
     3661 atttgactga tggcatgaca gttcgtgaac tgtgtagtgc cgctattacg atgtctgata
     3721 acaccgccgc caaccttctg cttacgacca ttggcggtcc gaaggagctg acggcattcc
     3781 ttcacaacat gggcgatcac gtcactcgtt tagaccgctg ggagccggaa ttaaacgaag
     3841 ctatccccaa tgacgaacgt gacacgacga tgcccgtcgc tatggccaca acgctgcgta
     3901 aattgctgac tggagagctt ttaaccctgg catctcgcca gcaacttatc gattggatgg
     3961 aggcagataa agtggccggg ccgctgctgc gctctgccct gccagccgga tggtttattg
     4021 cggataagtc tggggcggga gaacgcggtt ctcgtggcat tattgcagca cttggtccgg
     4081 atgggaaacc ctcacgtatt gtggtaatct atacgacagg ctcccaggcg acgatggacg
     4141 agcgtaaccg tcagatcgca gaaatcgggg cttctcttat taagcattgg taagcttcaa
     4201 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     4261 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata
     4321 taacttcgta taatgtatgc tatacgaagt tatccctcgg aggttccggt ggaagcggag
     4381 gtagcggcgg cagcgctgtc atgagacgaa ttctgacacg
//
