LOCUS       sequence-41124-e        6788 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 6788)
  TITLE     James McNally Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6788)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pTSK450
FEATURES             Location/Qualifiers
     source          1..6788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     13..29
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     14..30
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        70..420
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     CDS             796..816
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             856..1566
                     /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(1001..1019)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1269..1288
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1573..2247
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNS*GTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     primer_bind     complement(2457..2476)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2458..2476)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2483..2500)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2483..2499)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2492..2514)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2644..3099
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2731..2750)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2941..2962
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3360..4019)
                     /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(4020..4207)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(4352..4371)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4471..4493
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4531..4549)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4607..4711
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4712..4780,4781..5572)
                     /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(4930..4949)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5743..6331
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6485..6502
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(6655..6660,6661..6678,6679..6685)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6693..6709
                     /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     6698..6720
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6717..6733
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6717..6733
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6754..6772
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ccgggccccc cctcgaggtc gacggtatcg atatcttcgg atgcaagggt tcgaatccct
       61 tagctctcat tattttttgc tttttctctt gaggtcacat gatcgcaaaa tggcaaatgg
      121 cacgtgaagc tgtcgatatt ggggaactgt ggtggttggc aaatgactaa ttaagttagt
      181 caaggcgcca tcctcatgaa aactgtgtaa cataataacc gaagtgtcga aaaggtggca
      241 ccttgtccaa ttgaacacgc tcgatgaaaa aaataagata tatataaggt taagtaaagc
      301 gtctgttaga aaggaagttt ttcctttttc ttgctctctt gtcttttcat ctactatttc
      361 cttcgtgtaa tacagggtcg tcagatacat agatacaatt ctattacccc catccataca
      421 ggatcccgaa tggcttctaa ctttactcag ttcgttctcg tcgacaatgg cggaactggc
      481 gacgtgactg tcgccccaag caacttcgct aacgggatcg ctgaatggat cagctctaac
      541 tcgcgttcac aggcttacaa agtaacctgt agcgttcgtc agagctctgc gcagaatcgc
      601 aaatacacca tcaaagtcga ggtgcctaaa ggcgcctggc gttcgtactt aaatatggaa
      661 ctaaccattc caattttcgc cacgaattcc gactgcgagc ttattgttaa ggcaatgcaa
      721 ggtctcctaa aagatggaaa cccgattccc tcagcaatcg cagcaaactc cggcatctac
      781 gatgagctgt acaagccaaa aaagaagaga aaggtagcgg ccgccgctgc tgccgctgca
      841 gctgctgcag ctgcgatggt gagcaagggc gaggaggata acatggccat catcaaggag
      901 ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag
      961 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     1021 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag
     1081 gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc
     1141 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac
     1201 tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc
     1261 tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg
     1321 taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc
     1381 ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
     1441 ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc
     1501 gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac
     1561 aagtaaccgc ggatgtctgt tattaatttc acaggtagtt ctggtccatt ggtgaaagtt
     1621 tgcggcttgc agagcacaga ggccgcagaa tgtgctctag attccgatgc tgacttgctg
     1681 ggtattatat gtgtgcccaa tagaaagaga acaattgacc cggttattgc aaggaaaatt
     1741 tcaagtcttg taaaagcata taaaaatagt taaggcactc cgaaatactt ggttggcgtg
     1801 tttcgtaatc aacctaagga ggatgttttg gctctggtca atgattacgg cattgatatc
     1861 gtccaactgc atggagatga gtcgtggcaa gaataccaag agttcctcgg tttgccagtt
     1921 attaaaagac tcgtatttcc aaaagactgc aacatactac tcagtgcagc ttcacagaaa
     1981 cctcattcgt ttattccctt gtttgattca gaagcaggtg ggacaggtga acttttggat
     2041 tggaactcga tttctgactg ggttggaagg caagagagcc ccgaaagctt acattttatg
     2101 ttagctggtg gactgacgcc agaaaatgtt ggtgatgcgc ttagattaaa tggcgttatt
     2161 ggtgttgatg taagcggagg tgtggagaca aatggtgtaa aagactctaa caaaatagca
     2221 aatttcgtca aaaatgctaa gaaataggtt attactgagt agtatttatt taagtattgt
     2281 ttgtgcactt gcctgcaggc cttttgaaaa gcaagcataa aagatctaaa cataaaatct
     2341 gtaaaataac aagatgtaaa gataatgcta aatcatttgg ctttttgatt gattgtacag
     2401 gaaaatatac atcgcagggg gttgactttt accatttcac cgcgagctcc aattcgccct
     2461 atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     2521 ctggcgttac ccaacttaat cgccttgcag cacatccccc cttcgccagc tggcgtaata
     2581 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
     2641 gcgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2701 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2761 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2821 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     2881 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2941 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3001 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3061 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc ctgatgcggt attttctcct
     3121 tacgcatctg tgcggtattt cacaccgcat atgatccgtc gagttcaaga gaaaaaaaaa
     3181 gaaaaagcaa aaagaaaaaa ggaaagcgcg cctcgttcag aatgacacgt atagaatgat
     3241 gcattacctt gtcatcttca gtatcatact gttcgtatac atacttactg acattcatag
     3301 gtatacatat atacacatgt atatatatcg tatgctgcag ctttaaataa tcggtgtcac
     3361 tacataagaa cacctttggt ggagggaaca tcgttggtac cattgggcga ggtggcttct
     3421 cttatggcaa ccgcaagagc cttgaacgca ctctcactac ggtgatgatc attcttgcct
     3481 cgcagacaat caacgtggag ggtaattctg ctagcctctg caaagctttc aagaaaatgc
     3541 gggatcatct cgcaagagag atctcctact ttctcccttt gcaaaccaag ttcgacaact
     3601 gcgtacggcc tgttcgaaag atctaccacc gctctggaaa gtgcctcatc caaaggcgca
     3661 aatcctgatc caaacctttt tactccacgc gccagtaggg cctctttaaa agcttgaccg
     3721 agagcaatcc cgcagtcttc agtggtgtga tggtcgtcta tgtgtaagtc accaatgcac
     3781 tcaacgatta gcgaccagcc ggaatgcttg gccagagcat gtatcatatg gtccagaaac
     3841 cctatacctg tgtggacgtt aatcacttgc gattgtgtgg cctgttctgc tactgcttct
     3901 gcctcttttt ctgggaagat cgagtgctct atcgctaggg gaccaccctt taaagagatc
     3961 gcaatctgaa tcttggtttc atttgtaata cgctttacta gggctttctg ctctgtcatc
     4021 tttgccttcg tttatcttgc ctgctcattt tttagtatat tcttcgaaga aatcacatta
     4081 ctttatataa tgtataattc attatgtgat aatgccaatc gctaagaaaa aaaaagagtc
     4141 atccgctagg tggaaaaaaa aaaatgaaaa tcattaccga ggcataaaaa aatatagagt
     4201 gtactagagg aggccaagag taatagaaaa agaaaattgc gggaaaggac tgtgttatga
     4261 cttccctgac taatgccgtg ttcaaacgat acctggcagt gactcctagc gctcaccaag
     4321 ctcttaaaac ggaattatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4381 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     4441 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4501 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4561 atgtcatgat aataagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4621 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4681 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4741 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4801 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4861 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4921 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     4981 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5041 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5101 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5161 tcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5221 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5281 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5341 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5401 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5461 tggtaagccc tcccgtatcg tagttatcta cacgacgggc agtcaggcaa ctatggatga
     5521 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5581 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5641 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5701 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5761 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5821 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5881 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5941 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6001 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6061 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6121 cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6181 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag gggggaacgc
     6241 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6301 atgctcgtca ggggggccga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6361 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6421 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     6481 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     6541 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     6601 cagtgagcgc aacgcaatta atgtgagtta cctcactcat taggcacccc aggctttaca
     6661 ctttatgctt ccggctccta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     6721 aaacagctat gaccatgatt acgccaagct cggaattaac cctcactaaa gggaacaaaa
     6781 gctgggta
//
