LOCUS       sequence-361214-        5449 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dominate negative mCherry Rab11a in mammalian cells and 
            zebrafish embryos.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5449)
  AUTHORS   Krishnan N, Swoger M, Rathbun LI, Fioramonti PJ, Freshour J, Bates 
            M, Patteson AE, Hehnly H
  TITLE     Rab11 endosomes and Pericentrin coordinate centrosome movement 
            during pre-abscission in vivo.
  JOURNAL   Life Sci Alliance. 2022 Mar 18;5(7). pii: 5/7/e202201362. doi: 
            10.26508/lsa.202201362. Print 2022 Jul.
  PUBMED    35304423
REFERENCE   2  (bases 1 to 5449)
  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: "Life Sci 
            Alliance."; date: "2022-03-18"; volume: "5(7). pii: 5/7/e202201362. 
            doi"; pages: " 10.26508/lsa.202201362. Print 2022 Jul"
COMMENT     Sequence Label: pCS2-mCherry-Rab11a (S25N)
FEATURES             Location/Qualifiers
     source          1..5449
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      84..93
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             90..797
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(235..253)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     503..522
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1501..1635
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1525..1544)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1579..1598
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1748..1768)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1748..1766)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1787..1803)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1787..1803)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1800..1822)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1811..1827
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1835..1841,1842..1859,1860..1865))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1880..1901
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2018..2035)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2189..2777)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2269..2288)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2948..3739,3740..3808))
                     /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     3571..3590
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3809..3913)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3939..4394)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4076..4097)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4288..4307
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4465..5449
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg gtcgccacca tggtgagcaa gggcgaggag gataacatgg
      121 ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg
      181 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
      241 tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca
      301 tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt
      361 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga
      421 ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg
      481 gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct
      541 cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga
      601 agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc
      661 ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg
      721 aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca
      781 tggacgagct gtacaagaag ggcacccgcg acgacgagta cgactacctc ttcaaagttg
      841 tccttattgg agattctggt gttggaaaaa ataatcttct atctcgattt actcgaaatg
      901 agtttaatct ggaaagcaag agcaccattg gagtagaatt tgcaacaaga agcatccagg
      961 ttgatgggaa aacaataaaa gcacagatat gggacacggc agggcaagag cgataccgag
     1021 ctataacatc agcatactat cgtggagctg taggtgcctt attggtttat gacattgcta
     1081 aacatctcac atatgaaaat gtagagcggt ggctgaaaga actgagagat catgcggata
     1141 gtaacattgt tatcatgctc gtgggtaaca agagtgattt acgtcatctc agggcagttc
     1201 ctactgatga agcaagagct tttgcagaaa agaatggttt gtcatttatt gagacatctg
     1261 ctttagattc tacaaatgta gaagctgctt ttcaaacaat tctaacagag atataccgca
     1321 ttgtttctca gaagcagatg tcagacagac gtgagaatga catgtctcca agcaacaatg
     1381 tggttcctat tcatgttcca ccaaccactg aaaacaagcc aaaggtgcag tgctgtcaga
     1441 acatctaaga attcgccagc acagtctgca gtctagaact atagtgagtc gtattacgta
     1501 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     1561 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     1621 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     1681 gtgtgggagg ttttttaatt cgcggccgcg gcgccaatgc attgggcccg gtacccagct
     1741 tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc
     1801 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccggg agcataaagt
     1861 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     1921 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     1981 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2041 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2101 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2161 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2221 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2281 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2341 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2401 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2461 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2521 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2581 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2641 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2701 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2761 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2821 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2881 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2941 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3001 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3061 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3121 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3181 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3241 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3301 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3361 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3421 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3481 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3541 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3601 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3661 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3721 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3781 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     3841 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3901 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     3961 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     4021 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     4081 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     4141 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     4201 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     4261 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     4321 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     4381 gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     4441 gcgggcctct tcgctattac gccagtcgac cgccaattca atatggcgta tatggactca
     4501 tgccaattca atatggtgga tctggacctg tgccaattca atatggcgta tatggactcg
     4561 tgccaattca atatggtgga tctggacccc agccaattca atatggcgga cttggcacca
     4621 tgccaattca atatggcgga cctggcactg tgccaactgg ggaggggtct acttggcacg
     4681 gtgccaagtt tgaggagggg tcttggccct gtgccaagtc cgccatattg aattggcatg
     4741 gtgccaataa tggcggccat attggctata tgccaggatc aatatatagg caatatccaa
     4801 tatggcccta tgccaatatg gctattggcc aggttcaata ctatgtattg gccctatgcc
     4861 atatagtatt ccatatatgg gttttcctat tgacgtagat agcccctccc aatgggcggt
     4921 cccatatacc atatatgggg cttcctaata ccgcccatag ccactccccc attgacgtca
     4981 atggtctcta tatatggtct ttcctattga cgtcatatgg gcggtcctat tgacgtatat
     5041 ggcgcctccc ccattgacgt caattacggt aaatggcccg cctggctcaa tgcccattga
     5101 cgtcaatagg accacccacc attgacgtca atgggatggc tcattgccca ttcatatccg
     5161 ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc cacttggcag tacatcaata
     5221 tctattaata gtaacttggc aagtacatta ctattggaag tacgccaggg tacattggca
     5281 gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag tacatcccca
     5341 ttgacgtcaa tggggagggg caatgacgca aatgggcgtt ccattgacgt aaatgggcgg
     5401 taggcgtgcc taatgggagg tctatataag caatgctcgt ttagggaac
//
