LOCUS       sequence-47370-e        7667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7667)
  AUTHORS   Markus SM, Punch JJ, Lee WL
  TITLE     Motor- and tail-dependent targeting of dynein to microtubule plus 
            ends and the cell cortex.
  JOURNAL   Curr Biol. 2009 Feb 10;19(3):196-205. Epub 2009 Jan 29.
  PUBMED    19185494
REFERENCE   2  (bases 1 to 7667)
  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: "Curr 
            Biol."; date: "2009-02-10"; volume: "19(3)"; pages: "196-205. Epub 
            2009 Jan 29"
COMMENT     Sequence Label: pRS305:3mCherry
FEATURES             Location/Qualifiers
     source          1..7667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2243..2261
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2290..2306
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2291..2307
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2358..3065
                     /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"
     CDS             3072..3779
                     /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"
     CDS             3792..4502
                     /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(4504..4520)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4504..4520)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4552..4571)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4553..4571)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4578..4595)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4578..4594)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4587..4609)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4739..5194
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4826..4845)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5036..5057
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5494..5898
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5911..7005
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(7498..7517)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7617..7639
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttt cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggca gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg ggggaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggccgag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttac ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcctat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctc ggaattaacc ctcactaaag ggaacaaaag ctgggtaccg
     2281 ggccccccct cgaggtcgac ggtatcgata agcttgatat cgaattcctg cagcccgggg
     2341 gatccgctcc aagtggaatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg
     2401 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
     2461 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     2521 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
     2581 aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg
     2641 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg
     2701 actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc
     2761 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga
     2821 tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg
     2881 gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
     2941 ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca
     3001 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
     3061 acaagcctag gatggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca
     3121 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
     3181 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
     3241 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
     3301 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
     3361 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
     3421 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
     3481 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
     3541 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     3601 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     3661 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     3721 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaaga
     3781 ctaggagatc catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca
     3841 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
     3901 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
     3961 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
     4021 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
     4081 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
     4141 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
     4201 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
     4261 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     4321 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     4381 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     4441 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt
     4501 agagatccac tagttctaga gcggccgcca ccgcggtgga gctccaattc gccctatagt
     4561 gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     4621 gttacccaac ttaatcgcct tgcagcacat ccccccttcg ccagctggcg taatagcgaa
     4681 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcgac
     4741 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4801 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4861 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4921 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4981 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5041 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5101 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5161 gcgaatttta acaaaatatt aacgtttaca atttcctgat gcggtatttt ctccttacgc
     5221 atctgtgcgg tatttcacac cgcatatcga ccctcgagga gaacttctag tatatccaca
     5281 tacctaatat tattgcctta ttaaaaatgg aatcccaaca attacatcaa aatccacatt
     5341 ctcttcaaaa tcaattgtcc tgtacttcct tgttcatgtg tgttcaaaaa cgttatattt
     5401 ataggataat tatactctat ttctcaacaa gtaattggtt gtttggccga gcggtctaag
     5461 gcgcctgatt caagaaatat cttgaccgca gttaactgtg ggaatactca ggtatcgtaa
     5521 gatgcaagag ttcgaatctc ttagcaacca ttattttttt cctcaacata acgagaacac
     5581 acaggggcgc tatcgcacag aatcaaattc gatgactgga aattttttgt taatttcaga
     5641 ggtcgcctga cgcatatacc tttttcaact gaaaaattgg gagaaaaagg aaaggtgaga
     5701 ggccggaacc ggcttttcat atagaataga gaagcgttca tgactaaatg cttgcatcac
     5761 aatacttgaa gttgacaata ttatttaagg acctattgtt ttttccaata ggtggttagc
     5821 aatcgtctta ctttctaact tttcttacct tttacatttc agcaatatat atatatattt
     5881 caaggatata ccattctaat gtctgcccct atgtctgccc ctaagaagat cgtcgttttg
     5941 ccaggtgacc acgttggtca agaaatcaca gccgaagcca ttaaggttct taaagctatt
     6001 tctgatgttc gttccaatgt caagttcgat ttcgaaaatc atttaattgg tggtgctgct
     6061 atcgatgcta caggtgtccc acttccagat gaggcgctgg aagcctccaa gaaggttgat
     6121 gccgttttgt taggtgctgt gggtggtcct aaatggggta ccggtagtgt tagacctgaa
     6181 caaggtttac taaaaatccg taaagaactt caattgtacg ccaacttaag accatgtaac
     6241 tttgcatccg actctctttt agacttatct ccaatcaagc cacaatttgc taaaggtact
     6301 gacttcgttg ttgtcagaga attagtggga ggtatttact ttggtaagag aaaggaagac
     6361 gatggtgatg gtgtcgcttg ggatagtgaa caatacaccg ttccagaagt gcaaagaatc
     6421 acaagaatgg ccgctttcat ggccctacaa catgagccac cattgcctat ttggtccttg
     6481 gataaagcta atgttttggc ctcttcaaga ttatggagaa aaactgtgga ggaaaccatc
     6541 aagaacgaat tccctacatt gaaggttcaa catcaattga ttgattctgc cgccatgatc
     6601 ctagttaaga acccaaccca cctaaatggt attataatca ccagcaacat gtttggtgat
     6661 atcatctccg atgaagcctc cgttatccca ggttccttgg gtttgttgcc atctgcgtcc
     6721 ttggcctctt tgccagacaa gaacaccgca tttggtttgt acgaaccatg ccacggttct
     6781 gctccagatt tgccaaagaa taaggttgac cctatcgcca ctatcttgtc tgctgcaatg
     6841 atgttgaaat tgtcattgaa cttgcctgaa gaaggtaagg ccattgaaga tgcagttaaa
     6901 aaggttttgg atgcaggtat cagaactggt gatttaggtg gttccaacag taccaccgaa
     6961 gtcggtgatg ctgtcgccga agaagttaag aaaatccttg cttaaaaaga ttctcttttt
     7021 ttatgatatt tgtacataaa ctttataaat gaaattcata atagaaacga cacgaaatta
     7081 caaaatggaa tatgttcata gggtagacga aactatatac gcaatctaca tacatttatc
     7141 aagaaggaga aaaaggagga tagtaaagga atacaggtaa gcaaattgat actaatggct
     7201 caacgtgata aggaaaaaga attgcacttt aacattaata ttgacaagga ggagggcacc
     7261 acacaaaaag ttaggtgtaa cagaaaatca tgaaactacg attcctaatt tgatattgga
     7321 ggattttctc taaaaaaaaa aaaatacaac aaataaaaaa cactcaatga cctgaccatt
     7381 tgatggagtt taagtcaata ccttcttgaa ccatttccca taatggtgaa agttccctca
     7441 agaattttac tctgtcagaa acggccttac gacgtagtcg atatggtgca ctctcagtac
     7501 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     7561 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     7621 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcga
//
