LOCUS       sequence-216270-        6546 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses myc tagged Miro2 T18N dominant negative mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6546)
  AUTHORS   Fransson S, Ruusala A, Aspenstrom P
  TITLE     The atypical Rho GTPases Miro-1 and Miro-2 have essential roles in 
            mitochondrial trafficking.
  JOURNAL   Biochem Biophys Res Commun. 2006 Jun 2;344(2):500-10. Epub 2006 Apr 
            3.
  PUBMED    16630562
REFERENCE   2  (bases 1 to 6546)
  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: "Biochem 
            Biophys Res Commun."; date: "2006-06-2"; volume: "344(2)"; pages: 
            "500-10. Epub 2006 Apr 3"
COMMENT     Sequence Label: pRK5-myc-Miro2 T18N
FEATURES             Location/Qualifiers
     source          1..6546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(33..38,39..56,57..63)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    71..87
                     /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)."
     primer_bind     76..98
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     95..111
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     95..111
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        140..519
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        520..723
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     674..694
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     720..744
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        954..972
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     954..971
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     985..1004
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1051..1060
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1060..1089
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      2806..2815
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2972..2981
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2985..3119
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3022..3041)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3076..3095
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3188..3545
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3188..3379
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3211..3231)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3396..3531
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3565..3582)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3565..3581)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3574..3596)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3794..4249
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3881..3900)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4091..4112
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4266..4285)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4385..4407
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4445..4463)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4531..4635
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4636..4704,4705..5496)
                     /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(4854..4873)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5667..6255
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6156..6175
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6409..6426
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    join(6543..6546,1..18)
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
       61 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg
      121 aattaattcg agctcgcccg acattgatta ttgactagtt attaatagta atcaattacg
      181 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      241 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      301 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      361 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      421 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      481 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
      541 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
      601 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
      661 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
      721 gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat
      781 agaagacacc gggaccgatc cagcctccgc ggccgggaac ggtgcattgg aacgcggatt
      841 ccccgtgcca agagtgacgt aagtaccgcc tatagagtct ataggcccac ccccttggct
      901 tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg
      961 tgacactata gaataacatc cactttgcct ttctctccac aggtgtccac tcccaggtcc
     1021 aactgcacct cggttctatc gattgaattg gccaccatgg agcagaagct gatctccgag
     1081 gaggacctgg gatctatgag gcgggacgtg cgcatcctgt tactgggcga ggcccaggtg
     1141 gggaagaact cgctgatcct gtccctggtg ggcgaggagt tccccgagga ggtccctccc
     1201 cgcgcggagg agatcaccat ccccgcggac gtcaccccgg agaaggtgcc cacccacatc
     1261 gtggactact cagaagccga gcagacggac gaggagctgc gggaggagat ccacaaggca
     1321 aacgtggtgt gtgtggtgta tgacgtctct gaggaggcca ccattgagaa gattcgaact
     1381 aagtggatcc cactggtgaa tggggggacc acgcaggggc ccagggtgcc catcatccta
     1441 gtgggcaaca agtcagacct gcggtcgggg agctccatgg aggccgtgct ccccatcatg
     1501 agccagtttt ccgagattga gacctgcgtg gagtgttcgg ccaagaacct gaggaacatc
     1561 tcagagctgt tctactacgc ccagaaggcc gtcctgcatc ccacagcccc cctctatgac
     1621 cctgaggcca agcagttgag gcccgcgtgc gcccaggcgc tgacgcgcat cttcaggctc
     1681 tcagatcagg acctggacca ggcgctcagt gacgaagagc tcaacgcttt ccagaaatcc
     1741 tgctttgggc accccctggc cccgcaggcc ctggaggacg tgaagacggt ggtgtgcagg
     1801 aacgtggcgg gcggcgtgcg ggaggaccag ctgaccctgg atggtttcct cttcctgaac
     1861 acgctcttca tccagcgcgg ccggcacgag accacctgga ccatcctgcg gcgcttcggc
     1921 tacagcgatg ccctggagct gactgcggac tatctctccc ctctgatcca cgtgcccccc
     1981 ggctgcagca cggagctcaa ccaccttggc taccagtttg tgcagagagt gtttgagaag
     2041 cacgaccagg accgcgacgg cgccctctcg cccgtggagc tgcaaagcct tttcagtgtg
     2101 ttcccagcag cgccctgggg ccccgagctc ccacgcacag tccgcacaga ggccggccgg
     2161 ttgcccctgc acggatacct ctgccagtgg accctggtga cctacctgga cgtccggagc
     2221 tgccttggac acctaggcta cctgggctac cccaccctct gtgagcagga ccaggcccat
     2281 gccatcacag tcactcgtga gaagaggctg gaccaggaga agggacagac gcagcggagc
     2341 gtcctcctgt gcaaggtggt aggggcctgt ggagtgggca agtctgcctt cctgcaggcc
     2401 tttctcggcc gcggcctggg gcaccaggac acgagggagc agcctcccgg ctacgccatc
     2461 gacacggtgc aggtcaatgg gcaggagaag tacttgatcc tctgtgaggt gggcacagat
     2521 ggtctgctgg ccacatcgct ggacgccacc tgtgacgttg cctgcttgat gtttgatggc
     2581 agtgacccaa agtcctttgc acattgtgcc agcgtctaca agcaccatta catggacggg
     2641 cagaccccct gcctctttgt ctcctccaag gccgacctgc ccgaaggtgt cgcggtgtct
     2701 ggcccatcac cggccgagtt ttgccgcaag caccggctac ccgctcccgt gccgttctcc
     2761 tgtgctggcc cagccgagcc cagcaccacc atcttcaccc agctcgccac catggccgcc
     2821 ttcccacatt tggtccacgc agagctgcat ccctcttcct tctggctccg ggggctgctg
     2881 ggggttgtcg gggccgccgt ggccgcagtc ctcagcttct cactctacag ggtcctggtg
     2941 aagagccagt gagaattcct gcagaagctt ggccgccatg gcccaacttg tttattgcag
     3001 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3061 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcg
     3121 ggaattaatt cggcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag
     3181 gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3241 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3301 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3361 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3421 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3481 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3541 gcaaaaagct gttaacagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa
     3601 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     3661 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     3721 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     3781 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3841 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3901 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3961 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     4021 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4081 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat
     4141 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4201 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     4261 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     4321 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4381 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     4441 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     4501 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4561 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4621 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4681 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4741 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4801 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4861 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4921 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4981 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5041 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5101 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5161 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5221 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5281 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5341 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5401 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5461 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5521 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5581 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5641 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5701 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5761 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5821 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5881 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5941 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6001 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6061 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6121 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6181 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6241 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6301 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6361 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     6421 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     6481 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     6541 attaat
//
