pJK148-Pnmt1-GBP-mCherry(C)-leu1

LOCUS      pJK148-Pnmt1-GBP-mCherry(C)-leu1               8527 bp DNA     circular SYN 27-JUN-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8527)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             307..1421
                     /codon_start=1
                     /label=leu1+ ORF
                     /translation="MCAKKIVVLPGDHIGPEIVASALEVLKVVEKKRPELKLEFEEHKI
                     GGASIDAYGTPLTDETVKACLEADGVLLGAVGGPEWTNPNCRPEQGLLKLRKSMGVWAN
                     LRPCNFASKSLVKYSPLKPEIVEGVDFCVVRELTGGCYFGERTEDNGSGYAMDTWPYSL
                     EEVSRIARLAAWLAETSNPPAPVTLLDKANVLATSRLWRKTVAKIFKEEYPHLTLKTSS
                     LTPQPCFWSRALVHLTVLF*LTTCLVTLFQMRLLSFLVAWAFCLLPPFPVW*ENQKKRF
                     IVWLSPFTVALPISLARALLILLVQFYLLPFSFVMV*MLLRRLKLSKPPYARSWMILQL
                     VDVVFILAIWEVRLLPLILLRLLLKNLKKFC"
     rep_origin      complement(2303..2758)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     2899..2915
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2925..2943
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2989..4147
                     /label=nmt1 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring medium strength thiamine-repressible expression"
     CDS             4182..4535
                     /codon_start=1
                     /label=GBP
                     /translation="MADVQLVESGGALVQPGGSLRLSCAASGFPVNRYSMRWYRQAPGK
                     EREWVAGMSSAGDRSSYEDSVKGRFTISRDDARNTVYLQMNSLKPEDTAVYYCNVNVGF
                     EYWGQGTQVTVSSS"
     CDS             4536..5246
                     /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
                     EGRHSTGAMDELYK"
     misc_feature    5247..6246
                     /label=nmt terminator
     promoter        complement(6267..6285)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6306..6322)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6330..6346
                     /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(6354..6384)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(6708..7296)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7467..8327)
                     /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"
     promoter        complement(8328..8432)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatacgat atcccaatct gtagtataaa aagtgcaaca gtgttgagtt tcccgaaacc
      241 aggcattttt gcggtagggt ctgaaacttt cacgaaacca tttcacaaca aaacttcttc
      301 aacaacatgt gtgcaaagaa aatcgtcgtc ttaccaggag accatattgg ccctgaaatt
      361 gttgcttctg ccttggaggt tttgaaagtc gttgagaaga agcgacctga gttaaaactc
      421 gagtttgaag aacacaagat tggaggtgcc tctattgatg cctatggaac ccctttgact
      481 gacgagactg tgaaggcttg tttggaagct gacggtgttc ttttgggtgc cgttggtggt
      541 cctgaatgga ccaaccccaa ttgtcgtcct gagcaaggtt tattgaagct tcgtaagagt
      601 atgggtgttt gggccaacct tcgaccttgc aactttgcca gcaagtcttt agtcaagtac
      661 agccctttga agcctgaaat cgttgaaggt gtcgattttt gtgttgtacg agaacttact
      721 ggaggttgtt actttggtga gcgcactgag gacaacggat cgggttatgc tatggacact
      781 tggccttaca gtttggaaga agtttctcgt attgctcgtt tggctgcttg gttagctgaa
      841 acttccaacc ctcctgctcc cgtcacatta ctcgacaaag ctaatgtttt ggcaacttct
      901 cgtctttggc gcaagactgt agctaagatt tttaaggagg aatatcctca ccttacctta
      961 aaaaccagct cattgactcc gcagccatgc ttttggtcaa gagccctcgt acacttaacg
     1021 gtgttgtttt gactgacaac ttgtttggtg acattatttc agatgaggct tctgtcattc
     1081 ctggtagctt gggccttttg ccttctgcct ccctttccgg tgtggtagga aaatcagaag
     1141 aaaaggttca ttgtttggtt gagcccattc acggtagcgc tcccgatatc gctggcaagg
     1201 gcattgttaa tcctgttggt acaattttat ctgcttccct tctccttcgt tatggtttga
     1261 atgctcctaa ggaggctgaa gctatcgaag ccgccgtacg caaggtcttg gatgatactt
     1321 caattggtgg acgtggtctt tatactcgcg atttgggagg tgaggcttct accgctgata
     1381 ttactaaggc tgttgttgaa gaacttgaaa aaattttgta atagggttta atgtagaata
     1441 aattcacatg atttgatttg acctaatttt tattgattaa ttgtgtcgca tatttattat
     1501 cgttaaaaat gcaattgaca aaagcgtaaa ttttaaggct ctaatgtttt ttcttttggt
     1561 atgttatagt caagcaaaga tgtattaagt tgcaaaagca tggaattgat ttacgacatc
     1621 cgttcaactt tatgaccgag ttgtcgctgt gtacttactc tttttttact atgaaaagtt
     1681 tcaaacataa tatcattaat tatttctttt taaacgaagt aacttgtaac ataaataata
     1741 ggggaaaaga agaaaaaaac cataaaaatt cgagaacgta acaattatta ttgtaacgat
     1801 gacaacccga ttctttgaga gagccagaga catgactttt gcattaccgt aattggtttc
     1861 tacttatgaa ttgtaaactt gtgtagggaa tttcatccac agctatcgcg ggctgctctc
     1921 tgcatcagtg ttagcacatt ttatattaat gaagattttg ctatgcaaaa tacgtacatt
     1981 tctggcgcat atatttttat caaacgaaca tgactatcct ttccactttg atggttcgca
     2041 agtattttga tttgtattaa atgcacatct aacaagccat ttcgcccaaa atcaccaact
     2101 ggcacatgta cccaaccagt tagttcctcg acttgaactg ttgtaacaat ttccaaatct
     2161 tgaaaccctg tcccagctga tattcgcaaa gtactgggag tataagattc atcaagggtg
     2221 tattgaaggt acatactcac atacttaatc gatgcggtgt gaaataccgc acagatgcgt
     2281 aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta
     2341 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     2401 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     2461 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     2521 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     2581 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     2641 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     2701 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     2761 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     2821 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     2881 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat
     2941 agggcgaatt ggctgcaggt cgatcgactc tagaggatca gaaaattatc gccataaaag
     3001 acagaataag tcatcagcgg ttgtttcatt tcctatattt tttttttatt tttttatttt
     3061 ttaataaggg aaaatttaac gtctaaggat acagaagatt gttagcacat taaagtaata
     3121 aaggcttaag tagtaagtgc cttagcatgt tattgtattt caaaggacat aatctaaaat
     3181 aataacaata tcatttctca caagttattc aattttcttt tttttttcta ataatatcaa
     3241 gaatgtatta tttgtttgac ataagtcaac taatttattt aatatgctgg attaatcttg
     3301 cagacatgta aattaacaag ttttagtcaa ataacgttga agtttcaatg aactcaaata
     3361 atttctcttt ttttttatat aaccataatc tgatttatat tttccgcagg gatcaactga
     3421 agttatgaca tttggattgg atcacttata accttggtcg ccaaataata caaaaatcag
     3481 cgttataaaa caaagaaggt ttttgttaag aaattaatcc tctttcttga taagaaagtt
     3541 gaaccgaaat tgcagatact gatatatgaa aataataccc acaattttgg gaatagcgca
     3601 agcctcaatt taaacaatag gtgaggacac atgataatga cctcaatgat tgttagaaga
     3661 aaagagcctc attacaaaat cgaaaaatga atggttgggt acaagtttcc aaaacatggt
     3721 aaagtggact ttgcgtatga gacgtaaata gaaaaaaaca cttgttatat gttttctaga
     3781 attattgttg tctctttatg gttggatgat gcaaaatagt aatttcggtt agttgctgta
     3841 aaacaccacg agacaaatag atatggatat ttattaaatc aggaaaaacg taactctcgg
     3901 ctactggatg gttcagtcac ccaacgatta ctggggagag aaaacagggc aaaagcaaag
     3961 cttaaaggaa tccgattgtc attcggcaat gtgcagcgaa actaaaaacc ggataatgga
     4021 cctgttaatc gaaacattga agatatataa aggaagagga atcctggcat atcatcaatt
     4081 gaataagttg aattaattat ttcaatctca ttctcacttt ctgacttata gtcgctttgt
     4141 taaatcatat gtcgacatgg taccagatct ctcgaggatc catggctgat gtccaactgg
     4201 tggagtctgg tggcgctttg gtgcagccag gtggctctct gcgtttgtcc tgtgccgctt
     4261 ctggcttccc agtgaaccgc tattccatgc gctggtatcg ccaggctcca ggcaaagagc
     4321 gtgagtgggt agccggtatg tccagcgcgg gtgatcgtag ctcctatgaa gactccgtga
     4381 agggccgttt caccatcagc cgtgacgatg cccgtaacac ggtgtatctg caaatgaaca
     4441 gcttgaaacc tgaagatacg gccgtgtatt actgtaatgt gaacgtgggc ttcgagtatt
     4501 ggggccaagg cacccaggtc accgtctcct ctagcatggt gagcaagggc gaggaggata
     4561 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     4621 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctatgagggc acccagaccg
     4681 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     4741 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     4801 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     4861 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     4921 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagaca atgggctggg
     4981 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
     5041 ggctgaagct gaaggacggt ggccactacg acgctgaggt caagaccacc tacaaggcca
     5101 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctctc
     5161 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     5221 gcgccatgga cgagctgtac aagtaaccat ggggtaaaag gaatgtctcc cttgccagta
     5281 ctgctagggt ttttctttca aactatggaa gcccattcaa gctgcatatt acgattttgt
     5341 ttttcgcttt tagaaagtgg tttagatgag ataatagaaa aattcttgat ctccgacaac
     5401 gagtactttt attttttttg ctaatcactt tactcaatat tagctcgaaa tcgtagaaac
     5461 gtagacgggt gcgggatacc gagtggtgta gttaagaatt tttataaacc acgtggccca
     5521 aaaatatgaa cccaaaacgt ttatacatga gtatacttta agaaggctat accccttcgt
     5581 gttagatgta gttttagcta cccaacccga gtctatgagc ttgacttcag atgtagaagg
     5641 cattaaatcg ttttgaatat taattaaaaa acgatgaaaa ttaaatattt aaaagcaatc
     5701 atacgctgaa aatttagtgc tgtggctaat ccttcaacat ggaaatgcca taaaagtgac
     5761 tttgacaaaa aaaaaagtat atacaggtag taaactcatc tacttcattg actttgttta
     5821 cagcatgtgg aaggaggaat atttattgct aaatcgtagt ttaacattca ataagtaata
     5881 ctattgaaat tcgacaagat tggccgcatg gatgaaaaag aggcattttg ctttgggaga
     5941 attagtccaa attagaactg aaaaaaaaaa ctttacgagg caaaaatgtc ggattgagat
     6001 cgtaaaagtt cgctcgtcgt cttttgcttt gtgattgttt tcatggatac atcttgctgg
     6061 atatttaaat tttagtacta tgtataagat attctataaa tgttttatca cccaaacctg
     6121 ttagcgcctt cttaattcta ttcaatctgg cttttgctct gagactactt cttggacttt
     6181 cactacttgt tagttatacg gaatttgtgt aattagaagt gaaataatcc tttctattag
     6241 taatgcgagc tccgcagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     6301 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     6361 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6421 attgcgttgc gcccactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6481 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6541 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6601 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6661 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6721 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6781 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6841 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6901 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6961 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     7021 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7081 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7141 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7201 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7261 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7321 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7381 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7441 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7501 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7561 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7621 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7681 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7741 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7801 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7861 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7921 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7981 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     8041 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     8101 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8161 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8221 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8281 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8341 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8401 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8461 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     8521 tttcgtc
//