LOCUS       sequence-47366-e        6606 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6606)
  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 6606)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pRS303:3mCherry
FEATURES             Location/Qualifiers
     source          1..6606
                     /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        86..190
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(191..259,260..1051)
                     /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(409..428)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1222..1810
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1964..1981
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2134..2139,2140..2157,2158..2164)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2172..2188
                     /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     2177..2199
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2196..2212
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2196..2212
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2233..2251
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2280..2296
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2281..2297
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2348..3055
                     /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             3062..3769
                     /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             3782..4492
                     /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(4494..4510)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4494..4510)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4542..4561)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4543..4561)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4568..4585)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4568..4584)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4577..4599)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4729..5184
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4816..4835)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5026..5047
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5445..6104)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(6105..6292)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(6437..6456)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6556..6578
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataagacgtc
       61 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
      121 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      181 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      241 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      301 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
      361 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
      421 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
      481 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
      541 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
      601 gacaacgatc ggaggaccga aggagctaac cgcttttttt cacaacatgg gggatcatgt
      661 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
      721 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
      781 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
      841 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
      901 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
      961 agttatctac acgacgggca gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     1021 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1081 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1141 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1201 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1261 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1321 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     1381 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1441 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1501 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1561 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga
     1621 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1681 aacaggagag cgcacgaggg agcttccagg ggggaacgcc tggtatcttt atagtcctgt
     1741 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggccgag
     1801 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     1861 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     1921 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     1981 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     2041 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     2101 tgtgagttac ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcctat
     2161 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     2221 cgccaagctc ggaattaacc ctcactaaag ggaacaaaag ctgggtaccg ggccccccct
     2281 cgaggtcgac ggtatcgata agcttgatat cgaattcctg cagcccgggg gatccgctcc
     2341 aagtggaatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
     2401 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     2461 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
     2521 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     2581 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     2641 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     2701 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     2761 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     2821 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     2881 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     2941 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     3001 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagcctag
     3061 gatggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     3121 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     3181 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     3241 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     3301 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     3361 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     3421 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     3481 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     3541 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     3601 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     3661 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     3721 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaaga ctaggagatc
     3781 catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     3841 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     3901 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     3961 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     4021 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     4081 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     4141 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     4201 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     4261 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     4321 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     4381 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     4441 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt agagatccac
     4501 tagttctaga gcggccgcca ccgcggtgga gctccaattc gccctatagt gagtcgtatt
     4561 acaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     4621 ttaatcgcct tgcagcacat ccccccttcg ccagctggcg taatagcgaa gaggcccgca
     4681 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcgac gcgccctgta
     4741 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4801 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4861 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4921 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4981 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     5041 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     5101 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5161 acaaaatatt aacgtttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg
     5221 tatttcacac cgcatatgat ccgtcgagtt caagagaaaa aaaaagaaaa agcaaaaaga
     5281 aaaaaggaaa gcgcgcctcg ttcagaatga cacgtataga atgatgcatt accttgtcat
     5341 cttcagtatc atactgttcg tatacatact tactgacatt cataggtata catatataca
     5401 catgtatata tatcgtatgc tgcagcttta aataatcggt gtcactacat aagaacacct
     5461 ttggtggagg gaacatcgtt ggtaccattg ggcgaggtgg cttctcttat ggcaaccgca
     5521 agagccttga acgcactctc actacggtga tgatcattct tgcctcgcag acaatcaacg
     5581 tggagggtaa ttctgctagc ctctgcaaag ctttcaagaa aatgcgggat catctcgcaa
     5641 gagagatctc ctactttctc cctttgcaaa ccaagttcga caactgcgta cggcctgttc
     5701 gaaagatcta ccaccgctct ggaaagtgcc tcatccaaag gcgcaaatcc tgatccaaac
     5761 ctttttactc cacgcgccag tagggcctct ttaaaagctt gaccgagagc aatcccgcag
     5821 tcttcagtgg tgtgatggtc gtctatgtgt aagtcaccaa tgcactcaac gattagcgac
     5881 cagccggaat gcttggccag agcatgtatc atatggtcca gaaaccctat acctgtgtgg
     5941 acgttaatca cttgcgattg tgtggcctgt tctgctactg cttctgcctc tttttctggg
     6001 aagatcgagt gctctatcgc taggggacca ccctttaaag agatcgcaat ctgaatcttg
     6061 gtttcatttg taatacgctt tactagggct ttctgctctg tcatctttgc cttcgtttat
     6121 cttgcctgct cattttttag tatattcttc gaagaaatca cattacttta tataatgtat
     6181 aattcattat gtgataatgc caatcgctaa gaaaaaaaaa gagtcatccg ctaggtggaa
     6241 aaaaaaaaat gaaaatcatt accgaggcat aaaaaaatat agagtgtact agaggaggcc
     6301 aagagtaata gaaaaagaaa attgcgggaa aggactgtgt tatgacttcc ctgactaatg
     6361 ccgtgttcaa acgatacctg gcagtgactc ctagcgctca ccaagctctt aaaacggaat
     6421 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     6481 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     6541 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     6601 gcgcga
//
