LOCUS       sequence-357179-        5044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor with homologous arms for Rab25 to knock in DExCon-mTagBFP2 
            module.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5044)
  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 5044)
  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: pJET-59-DExCon-mTagBFP2-R25
FEATURES             Location/Qualifiers
     source          1..5044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    7..25
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    43..61
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    79..97
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    115..133
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             292..1002
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     promoter        complement(1489..1507)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1813..1831)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1899..2003
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2004..2072,2073..2864)
                     /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(2222..2241)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3035..3623
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3524..3543
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3777..3794
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3914..3935
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3950..3955,3956..3973,3974..3980)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     primer_bind     4470..4492
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4484..4501
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4485..4501
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4512..4530
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4535..4546
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4568..4586
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4943..4961
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4979..4997
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5015..5033
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 gtttactccc tatcagtgat agagaacgta tgaccagttt actccctatc agtgatagag
       61 aacgtatcta cagtttactc cctatcagtg atagagaacg tatatccagt ttactcccta
      121 tcagtgatag agaacgtata agctttgctt atgtaaacca gggcgcctat aaaagagtgc
      181 tgattttttg agtaaacttc aattccacaa cacttttgtc ttataccaac tttccgtacc
      241 acttcctacc ctcgtaaaga attcaagatg gttggatcgg gtgggactag tatggtgtct
      301 aagggcgaag agctgattaa ggagaacatg cacatgaagc tgtacatgga gggcaccgtg
      361 gacaaccatc acttcaagtg cacatccgag ggcgaaggca agccctacga gggcacccag
      421 accatgagaa tcaaggtggt cgagggcggc cctctcccct tcgccttcga catcctggct
      481 actagcttcc tctacggcag caagaccttc atcaaccaca cccagggcat ccccgacttc
      541 ttcaagcagt ccttccctga gggcttcaca tgggagagag tcaccacata cgaagacggg
      601 ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat ctacaacgtc
      661 aagatcagag gggtgaactt cacatccaac ggccctgtga tgcagaagaa aacactcggc
      721 tgggaggcct tcaccgagac gctgtacccc gctgacggcg gcctggaagg cagaaacgac
      781 atggccctga agctcgtggg cgggagccat ctgatcgcaa acgccaagac cacatataga
      841 tccaagaaac ccgctaagaa cctcaagatg cctggcgtct actatgtgga ctacagactg
      901 gaaagaatca aggaggccaa caacgagacc tacgtcgagc agcacgaggt ggcagtggcc
      961 agatactgcg acctccctag caaactgggg cacaagctta ataccggtgc tggaagtggt
     1021 gggaatggaa ctgaggaaga ttataacttt gtcttcaagg gtgagttggg ttccctgaag
     1081 caagaggaag cctgagagac ccagaaagag attgaataga gccagagagg gccccctcct
     1141 gtttttttat ctcttaccaa gacctcctgg ggctcctcat tccttcagaa agaaagctgc
     1201 agggagtggg ggagggtctg gttgcagggg caaaagtcag ggagcttctg agtatgctct
     1261 actgtggtaa ataatgatgt tgattcctcc ttaagagtgc tgagttaggc tcctctgccc
     1321 cgttcctaat agaaacccaa atccctggtg tcagccagtg cagcacaaga gatggaggtc
     1381 tgggcctcat gggccttcct ttcactgccc gctttccagt cgggaaacct gtcgtgccag
     1441 atcttgctga aaaactcgag ccatccggaa gatctggcgg ccgctctccc tatagtgagt
     1501 cgtattacgc cggatggata tggtgttcag gcacaagtgt taaagcagtt gattttattc
     1561 actatgatga aaaaaacaat gaatggaacc tgctccaagt taaaaataga gataataccg
     1621 aaaactcatc gagtagtaag attagagata atacaacaat aaaaaaatgg tttagaactt
     1681 actcacagcg tgatgctact aattgggaca attttccaga tgaagtatca tctaagaatt
     1741 taaatgaaga agacttcaga gcttttgtta aaaattattt ggcaaaaata atataattcg
     1801 gctgcagggg cggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     1861 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1921 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1981 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     2041 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2101 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     2161 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     2221 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     2281 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2341 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2401 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2461 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     2521 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     2581 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     2641 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     2701 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     2761 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     2821 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     2881 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     2941 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3001 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3061 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3121 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3181 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3241 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3301 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3361 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3421 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3481 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3541 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3601 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3661 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     3721 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     3781 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     3841 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagca attggcagtg
     3901 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     3961 tgcttccggc tcgtataatg tgtggaatta tgagcggata ataatttcac acaggaggtt
     4021 taaactttaa acatgtcaaa agagacgtct tttgttaaga atgctgagga acttgcaaag
     4081 caaaaaatgg atgctattaa ccctgaactt tcttcaaaat ttaaattttt aataaaattc
     4141 ctgtctcagt ttcctgaagc ttgctctaaa cctcgttcaa aaaaaatgca gaataaagtt
     4201 ggtcaagagg aacatattga atatttagct cgtagttttc atgagagtcg attgccaaga
     4261 aaacccacgc cacctacaac ggttcctgat gaggtggtta gcatagttct taatataagt
     4321 tttaatatac agcctgaaaa tcttgagaga ataaaagaag aacatcgatt ttccatggca
     4381 gctgagaata ttgtaggaga tcttctagaa agatctggcg aaagggggat gtgctgcaag
     4441 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     4501 tgagcgcgac gtaatacgac tcactatagg gcgaattgaa ggaaggccgt caaggccgca
     4561 tagctgataa tacgactcac tatagggagc ccaggctgag agcagacacc caacctgtcg
     4621 aacctgtctg acgtcatcat ctctccaccc acctgggccc caggtctcca gccaccccgc
     4681 tcttcctgtt ctcagcttcc gtcctctctg cttccttaca gcacccccac ctgccagagc
     4741 tgatcctccc taggccctgc ctaaccttga gttggccccc aatccctctg gctgcagaag
     4801 tccccttacc cccaatgaga ggaggggcag gaccagatct tttgagagct gagggttgag
     4861 ggcattgagc caacacacag atttgtcgcc tctgtccccg aagacacctg caccctccat
     4921 gcggagccaa ggaattcttt actccctatc agtgatagag aacgtatgaa gagtttactc
     4981 cctatcagtg atagagaacg tatgcagact ttactcccta tcagtgatag agaacgtata
     5041 agga
//
