LOCUS       sequence-358132-        4956 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor with homologous arms for Rab11a to knock in DExogron 
            (=minilAA7-DExCon) mNeonGreen module.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4956)
  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 4956)
  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-49-DExogron-mNeonGreen-R11a
FEATURES             Location/Qualifiers
     source          1..4956
                     /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      1660..1669
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2496..2514)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2820..2838)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2906..3010
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3011..3079,3080..3871)
                     /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(3229..3248)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4042..4630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4531..4550
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4784..4801
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4921..4942
                     /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 ggactagtac gcgtggcttc
     1201 tctgagaccg tggacctgat gctgaacctg cagtccaata aggagggctc tgtggatctg
     1261 aagaacgtga gcgccgtgcc taaggagaag accacactga aggacccatc caagccccct
     1321 gccaaggcac aggtggtggg atggccaccc gtgcggaact acagaaagaa tatgatgacc
     1381 cagcagaaga caagctccac tagtggcgtg tctaagggcg aagaggacaa tatggccagc
     1441 ctgcctgcca cacacgagct gcacatcttc ggcagcatca acggcgtgga cttcgacatg
     1501 gtcggacaag gcaccggcaa tcccaacgac ggctacgagg aactgaacct gaagtccacc
     1561 aagggcgacc tgcagttcag cccctggatt ctggtgcctc acatcggcta cggcttccac
     1621 cagtacctgc cttatcctga cggcatgagc cctttccagg ccgccatggt tgatggctct
     1681 ggctaccagg tgcaccggac catgcagttt gaagatggcg ccagcctgac cgtgaactac
     1741 cggtacacat acgagggcag ccacatcaaa ggcgaggccc aagtgaaagg cacaggcttc
     1801 cctgctgatg gccccgtgat gaccaattct ctgacagccg ccgactggtg cagaagcaag
     1861 aaaacatacc ccaacgacaa gaccatcatc agcaccttca agtggtccta caccaccggc
     1921 aacggcaagc ggtacagaag caccgccaga accacctaca ccttcgccaa acctatggcc
     1981 gccaactacc tgaagaacca gcctatgtac gtgttccgca agaccgagct gaagcactcc
     2041 aagacagaac tgaacttcaa agagtggcag aaagccttca ccgacgtgat gggcatggac
     2101 gagctgtaca agggaaccgg tgctggaagt ggtacacgcg acgacgagta cgactacctc
     2161 tttaaaggtg aggccatggg ctctcgcact ctacacagtc ctcgttcggg gacccgggcc
     2221 actcccggtg gaccctcgtg ccggccaccc ctgcactgat ataggcctcc ctcagccctt
     2281 cctttttgtg cggttccgtc tcctacccag ctcagcctct tctcccccgc tcagacaggg
     2341 gtccccatca catgccgctc tctgagcgac ctctccatag gccttcgctg gcctcagagc
     2401 ccctccctgc gtgtccttcc cctggcggac tgccttctcc cacatcgatc ttgctgaaaa
     2461 actcgagcca tccggaagat ctggcggccg ctctccctat agtgagtcgt attacgccgg
     2521 atggatatgg tgttcaggca caagtgttaa agcagttgat tttattcact atgatgaaaa
     2581 aaacaatgaa tggaacctgc tccaagttaa aaatagagat aataccgaaa actcatcgag
     2641 tagtaagatt agagataata caacaataaa aaaatggttt agaacttact cacagcgtga
     2701 tgctactaat tgggacaatt ttccagatga agtatcatct aagaatttaa atgaagaaga
     2761 cttcagagct tttgttaaaa attatttggc aaaaataata taattcggct gcaggggcgg
     2821 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     2881 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2941 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3001 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3061 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3121 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3181 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3241 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3301 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3361 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3421 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3481 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3541 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3601 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3661 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3721 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3781 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3841 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3901 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3961 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4021 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4081 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4141 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4201 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4261 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4321 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4381 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4441 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4501 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4561 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4621 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4681 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4741 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4801 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     4861 atgcagctgg cacgacaggt ttcccgactg gaaagcaatt ggcagtgagc gcaacgcaat
     4921 taatgtgagt tagctcactc attaggcacc ccaggc
//
