LOCUS       sequence-357412-        4140 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor with homologous arms for Rab25 to knock in DExCon-mNeonGreen 
            module.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4140)
  AUTHORS   Gemperle J, Harrison TS, Flett C, Adamson AD, Caswell PT
  TITLE     On demand expression control of endogenous genes with DExCon, 
            DExogron and LUXon reveals differential dynamics of Rab11 family 
            members.
  JOURNAL   Elife. 2022 Jun 16;11. pii: 76651. doi: 10.7554/eLife.76651.
  PUBMED    35708998
REFERENCE   2  (bases 1 to 4140)
  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: "Elife."; 
            date: "2022-06-16"; volume: "11. pii: 76651. doi"; pages: " 
            10.7554/eLife.76651"
COMMENT     Sequence Label: pMKRQ_11_DExCon-mNeonGreen-R25
FEATURES             Location/Qualifiers
     source          1..4140
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     278..300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     292..309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     293..309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        320..338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             343..354
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        376..394
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    751..769
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    787..805
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    823..841
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    859..877
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    895..913
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    931..949
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    967..985
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      1399..1408
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2346..2363)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2456..3044)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2536..2555)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3218..4012)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4013..4117)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt
      181 gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      301 acggccagtg agcgcgacgt aatacgactc actatagggc gaattgaagg aaggccgtca
      361 aggccgcata gctgataata cgactcacta tagggagccc aggctgagag cagacaccca
      421 acctgtcgaa cctgtctgac gtcatcatct ctccacccac ctgggcccca ggtctccagc
      481 caccccgctc ttcctgttct cagcttccgt cctctctgct tccttacagc acccccacct
      541 gccagagctg atcctcccta ggccctgcct aaccttgagt tggcccccaa tccctctggc
      601 tgcagaagtc cccttacccc caatgagagg aggggcagga ccagatcttt tgagagctga
      661 gggttgaggg cattgagcca acacacagat ttgtcgcctc tgtccccgaa gacacctgca
      721 ccctccatgc ggagccaagg aattctttac tccctatcag tgatagagaa cgtatgaaga
      781 gtttactccc tatcagtgat agagaacgta tgcagacttt actccctatc agtgatagag
      841 aacgtataag gagtttactc cctatcagtg atagagaacg tatgaccagt ttactcccta
      901 tcagtgatag agaacgtatc tacagtttac tccctatcag tgatagagaa cgtatatcca
      961 gtttactccc tatcagtgat agagaacgta taagctttgc ttatgtaaac cagggcgcct
     1021 ataaaagagt gctgattttt tgagtaaact tcaattccac aacacttttg tcttatacca
     1081 actttccgta ccacttccta ccctcgtaaa gaattcaaga tggttggatc gggtgggact
     1141 agtggcgtgt ctaagggcga agaggacaat atggccagcc tgcctgccac acacgagctg
     1201 cacatcttcg gcagcatcaa cggcgtggac ttcgacatgg tcggacaagg caccggcaat
     1261 cccaacgacg gctacgagga actgaacctg aagtccacca agggcgacct gcagttcagc
     1321 ccctggattc tggtgcctca catcggctac ggcttccacc agtacctgcc ttatcctgac
     1381 ggcatgagcc ctttccaggc cgccatggtt gatggctctg gctaccaggt gcaccggacc
     1441 atgcagtttg aagatggcgc cagcctgacc gtgaactaca gatacacata cgagggcagc
     1501 cacatcaaag gcgaggccca agtgaaaggc acaggcttcc ctgctgatgg ccccgtgatg
     1561 accaattctc tgacagccgc cgactggtgc agaagcaaga aaacataccc caacgacaag
     1621 accatcatca gcaccttcaa gtggtcctac accaccggca acggcaagcg gtacagaagc
     1681 accgccagaa ccacctacac cttcgccaaa cctatggccg ccaactacct gaagaaccag
     1741 cctatgtacg tgttccgcaa gaccgagctg aagcactcca agacagaact gaacttcaaa
     1801 gagtggcaga aagccttcac cgacgtgatg ggcatggacg agctgtacaa gggaaccggt
     1861 gctggaagtg gtgggaatgg aactgaggaa gattataact ttgtcttcaa gggtgagttg
     1921 ggttccctga agcaagagga agcctgagag acccagaaag agattgaata gagccagaga
     1981 gggccccctc ctgttttttt atctcttacc aagacctcct ggggctcctc attccttcag
     2041 aaagaaagct gcagggagtg ggggagggtc tggttgcagg ggcaaaagtc agggagcttc
     2101 tgagtatgct ctactgtggt aaataatgat gttgattcct ccttaagagt gctgagttag
     2161 gctcctctgc cccgttccta atagaaaccc aaatccctgg tgtcagccag tgcagcacaa
     2221 gagatggagg tctgggcctc atgggccttc ctttcactgc ccgctttcca gtcgggaaac
     2281 ctgtcgtgcc agctgcatta acatggtcat agctgtttcc ttgcgtattg ggcgctctcc
     2341 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggg taaagcctgg ggtgcctaat
     2401 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2461 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2521 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2581 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2641 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2701 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2761 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2821 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2881 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2941 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3001 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3061 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3121 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3181 tctaaagtat atatgagtaa acttggtctg acagttatta gaaaaattca tccagcagac
     3241 gataaaacgc aatacgctgg ctatccggtg ccgcaatgcc atacagcacc agaaaacgat
     3301 ccgcccattc gccgcccagt tcttccgcaa tatcacgggt ggccagcgca atatcctgat
     3361 aacgatccgc cacgcccaga cggccgcaat caataaagcc gctaaaacgg ccattttcca
     3421 ccataatgtt cggcaggcac gcatcaccat gggtcaccac cagatcttcg ccatccggca
     3481 tgctcgcttt cagacgcgca aacagctctg ccggtgccag gccctgatgt tcttcatcca
     3541 gatcatcctg atccaccagg cccgcttcca tacgggtacg cgcacgttca atacgatgtt
     3601 tcgcctgatg atcaaacgga caggtcgccg ggtccagggt atgcagacga cgcatggcat
     3661 ccgccataat gctcactttt tctgccggcg ccagatggct agacagcaga tcctgacccg
     3721 gcacttcgcc cagcagcagc caatcacggc ccgcttcggt caccacatcc agcaccgccg
     3781 cacacggaac accggtggtg gccagccagc tcagacgcgc cgcttcatcc tgcagctcgt
     3841 tcagcgcacc gctcagatcg gttttcacaa acagcaccgg acgaccctgc gcgctcagac
     3901 gaaacaccgc cgcatcagag cagccaatgg tctgctgcgc ccaatcatag ccaaacagac
     3961 gttccaccca cgctgccggg ctacccgcat gcaggccatc ctgttcaatc atactcttcc
     4021 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4081 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
//
