LOCUS       sequence-358131-        4740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor with homologous arms for Rab11a to knock in DExCon-mNeonGreen 
            module.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4740)
  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 4740)
  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-22-DExCon-mNeonGreen-R11a
FEATURES             Location/Qualifiers
     source          1..4740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..24,25..31)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     promoter        472..490
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    807..825
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    843..861
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    879..897
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    915..933
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    951..969
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    987..1005
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1023..1041
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      1444..1453
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2280..2298)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2604..2622)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2690..2794
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2795..2863,2864..3655)
                     /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(3013..3032)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3826..4414
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4315..4334
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4568..4585
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4705..4726
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tttacacttt atgcttccgg ctcgtataat gtgtggaatt atgagcggat aataatttca
       61 cacaggaggt ttaaacttta aacatgtcaa aagagacgtc ttttgttaag aatgctgagg
      121 aacttgcaaa gcaaaaaatg gatgctatta accctgaact ttcttcaaaa tttaaatttt
      181 taataaaatt cctgtctcag tttcctgaag cttgctctaa acctcgttca aaaaaaatgc
      241 agaataaagt tggtcaagag gaacatattg aatatttagc tcgtagtttt catgagagtc
      301 gattgccaag aaaacccacg ccacctacaa cggttcctga tgaggtggtt agcatagttc
      361 ttaatataag ttttaatata cagcctgaaa atcttgagag aataaaagaa gaacatcgat
      421 tttccatggc agctgagaat attgtaggag atcttctaga aagatagctg ataatacgac
      481 tcactatagg gactatggta gctaggagtt ccaggactca gtttcccctt tgagcctcct
      541 ttagcgacta aagcttgaag ccccacgcat ctcgactctc gcgcacaccg cccttgttgg
      601 gctcaggggc ggggcgccgc ccccggaagt acttcccctt aaaggctggg gcctgccgga
      661 aatggcgcag cggcagggag gggctcttca cccagtccgg cagttgaagc tcggcgctcg
      721 ggttacccct gcagcgacgc cccctggtcc cacagatacc actgctgctc cgccctttcg
      781 ctcctcggcc gcgcagaatt ctttactccc tatcagtgat agagaacgta tgaagagttt
      841 actccctatc agtgatagag aacgtatgca gactttactc cctatcagtg atagagaacg
      901 tataaggagt ttactcccta tcagtgatag agaacgtatg accagtttac tccctatcag
      961 tgatagagaa cgtatctaca gtttactccc tatcagtgat agagaacgta tatccagttt
     1021 actccctatc agtgatagag aacgtataag ctttgcttat gtaaaccagg gcgcctataa
     1081 aagagtgctg attttttgag taaacttcaa ttccacaaca cttttgtctt ataccaactt
     1141 tccgtaccac ttcctaccct cgtaaagatg ggatcgggtg ggactagtgg cgtgtctaag
     1201 ggcgaagagg acaatatggc cagcctgcct gccacacacg agctgcacat cttcggcagc
     1261 atcaacggcg tggacttcga catggtcgga caaggcaccg gcaatcccaa cgacggctac
     1321 gaggaactga acctgaagtc caccaagggc gacctgcagt tcagcccctg gattctggtg
     1381 cctcacatcg gctacggctt ccaccagtac ctgccttatc ctgacggcat gagccctttc
     1441 caggccgcca tggttgatgg ctctggctac caggtgcacc ggaccatgca gtttgaagat
     1501 ggcgccagcc tgaccgtgaa ctaccggtac acatacgagg gcagccacat caaaggcgag
     1561 gcccaagtga aaggcacagg cttccctgct gatggccccg tgatgaccaa ttctctgaca
     1621 gccgccgact ggtgcagaag caagaaaaca taccccaacg acaagaccat catcagcacc
     1681 ttcaagtggt cctacaccac cggcaacggc aagcggtaca gaagcaccgc cagaaccacc
     1741 tacaccttcg ccaaacctat ggccgccaac tacctgaaga accagcctat gtacgtgttc
     1801 cgcaagaccg agctgaagca ctccaagaca gaactgaact tcaaagagtg gcagaaagcc
     1861 ttcaccgacg tgatgggcat ggacgagctg tacaagggaa ccggtgctgg aagtggtaca
     1921 cgcgacgacg agtacgacta cctctttaaa ggtgaggcca tgggctctcg cactctacac
     1981 agtcctcgtt cggggacccg ggccactccc ggtggaccct cgtgccggcc acccctgcac
     2041 tgatataggc ctccctcagc ccttcctttt tgtgcggttc cgtctcctac ccagctcagc
     2101 ctcttctccc ccgctcagac aggggtcccc atcacatgcc gctctctgag cgacctctcc
     2161 ataggccttc gctggcctca gagcccctcc ctgcgtgtcc ttcccctggc ggactgcctt
     2221 ctcccacatc gatcttgctg aaaaactcga gccatccgga agatctggcg gccgctctcc
     2281 ctatagtgag tcgtattacg ccggatggat atggtgttca ggcacaagtg ttaaagcagt
     2341 tgattttatt cactatgatg aaaaaaacaa tgaatggaac ctgctccaag ttaaaaatag
     2401 agataatacc gaaaactcat cgagtagtaa gattagagat aatacaacaa taaaaaaatg
     2461 gtttagaact tactcacagc gtgatgctac taattgggac aattttccag atgaagtatc
     2521 atctaagaat ttaaatgaag aagacttcag agcttttgtt aaaaattatt tggcaaaaat
     2581 aatataattc ggctgcaggg gcggcctcgt gatacgccta tttttatagg ttaatgtcat
     2641 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2701 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2761 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     2821 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     2881 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     2941 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3001 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3061 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3121 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3181 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3241 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3301 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     3361 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3421 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3481 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     3541 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     3601 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     3661 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3721 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3781 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3841 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3901 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3961 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4021 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4081 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4141 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4201 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4261 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4321 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4381 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4441 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4501 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     4561 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     4621 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     4681 aattggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
//
