pRS426_GFP_LplA

LOCUS       pRS426_GFP_LplA             8493 bp DNA     circular SYN 03-JUL-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8493)
  AUTHORS   Westlake University
  TITLE     Direct Submission
  JOURNAL   Exported Sep 30, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(604..1059)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     1200..1216
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1226..1244
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1270..1286
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1527..1813
                     /gene="S. cerevisiae TDH3"
                     /label=TDH3 promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     misc_feature    1818..2531
                     /label=GFP
     misc_feature    2532..2567
                     /label=linker
     CDS             2568..3611
                     /label=LplA
     CDS             3591..3608
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4098..4114)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4151..4169)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4190..4206)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4214..4230
                     /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(4238..4268)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4283..4304
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4592..5180)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5351..6211)
                     /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(6212..6316)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6343..7685
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(7306..7353)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        7947..8162
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             join(8163..8493,1..473)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
ORIGIN
        1 gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta
       61 gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag
      121 cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca
      181 ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa
      241 gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag
      301 acgcattggg tcaacagtat agaaccgtgg atgatgtggt ctctacagga tctgacatta
      361 ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt
      421 acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg
      481 tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc
      541 agttattacc ctatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
      601 aggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
      661 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
      721 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
      781 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
      841 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
      901 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
      961 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     1021 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg
     1081 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     1141 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     1201 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
     1261 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gaattcctgc agcccacagt
     1321 ttattcctgg catccactaa atataatgga gcccgctttt taagctggca tccagaaaaa
     1381 aaagaatccc agcaccaaaa tattgttttc ttcaccaacc atcagttcat aggtccattc
     1441 tcttagcgca actacagaga acaggggcac aaacaggcaa aaaacgggca caacctcaat
     1501 ggagtgatgc aacctgcctg gagtaaatga tgacacaagg caattgaccc acgcatgtat
     1561 ctatctcatt ttcttacacc ttctattacc ttctgctctc tctgatttgg aaaaagctga
     1621 aaaaaaaggt tgaaaccagt tccctgaaat tattccccta cttgactaat aagtatataa
     1681 agacggtagg tattgattgt aattctgtaa atctatttct taaacttctt aaattctact
     1741 tttatagtta gtcttttttt agttttaaaa caccaagaac ttagtttcga ataaacacac
     1801 ataaacaaac aaagatggtt tctaaaggtg aagaattgtt tactggtgtt gttccaattt
     1861 tggttgaatt ggatggtgat gttaatggtc ataaattttc tgtttccggt gaaggtgaag
     1921 gtgatgctac ttatagtaaa ttgactttga agtttatttg taccaccggt aaattgcctg
     1981 ttccatggcc aactttggtt actactttaa cttatggtgt tcaatgtttc tctagatatc
     2041 cagatcatat gaagcaacat gatttcttca aatctgctat gccaaaaggt tatgttcaag
     2101 aaagaactat tttcttcaag gatgatggta attacaagac cagagctgaa gttaagtttg
     2161 aaggtgatac tttagttaac agaattgaat tgaagggtat tgatttcaag gaagatggta
     2221 atattttggg tcataagttg gaatataact acaattctca taacgtttac attatggctg
     2281 ataaacaaaa gaacggtatt aaagttaact tcaagattag acataacatt gaagatggtt
     2341 cagttcaatt ggctgatcat tatcaacaaa acaccccaat tggtgatggt ccagttttgt
     2401 tgccagataa tcattacttg tctactcaat caaagttgtc aaaagatcca aatgaaaaga
     2461 gagatcatat ggttttattg gaatttgtta ccgctgctgg tattactttg ggtatggatg
     2521 aattgtacaa atctagagct gatggttctg gttccggttc tgagctcatg ggttcaactt
     2581 tgagattgtt gatttctgat tcttacgatc catggtttaa cttggctgtt gaagaatgta
     2641 ttttcagaca aatgccagct actcaaagag ttttgttttt gtggagaaat gctgatactg
     2701 ttgttattgg tagagcacaa aatccttgga aggaatgtaa tactagaaga atggaagaag
     2761 ataacgttag attggctaga agatcaagtg gtggtggtgc tgttttccat gatttaggta
     2821 atacatgttt cactttcatg gctggtaaac cagaatacga taaaacaatt tcaacctcta
     2881 ttgttttgaa cgctttgaat gctttgggtg ttagtgctga agctagtggt agaaatgatt
     2941 tggttgttaa aactgttgaa ggtgatagaa aagtttctgg ttctgcttat agagaaacta
     3001 aggatagagg ttttcatcat ggtactttgt tgttgaatgc tgatttgtcc agattggcta
     3061 attatttgaa tcctgataag aagaagttgg ctgctaaagg tattacttca gttagatcaa
     3121 gagttactaa cttaactgaa ttgttgcctg gtattactca tgaacaagtt tgtgaagcta
     3181 ttactgaagc atttttcgct cattatggtg aaagagttga agctgaaatt atttctccaa
     3241 ataagacccc agatttgcct aattttgctg aaacattcgc tagacaatct tcttgggaat
     3301 ggaattttgg tcaagctcca gcttttagtc atttgttgga tgaaagattt acctggggtg
     3361 gtgttgaatt gcattttgat gttgaaaagg gtcatattac aagagcacaa gttttcactg
     3421 attctttgaa tccagctcca ttggaagctc tagctggtag attgcaaggt tgtttgtata
     3481 gagctgatat gttgcaacaa gaatgtgaag cattattggt tgattttcca gaacaagaaa
     3541 aggaattgag agaattgagt gcttggatgg ctggtgcagt tagagaattc catcatcatc
     3601 atcatcatta actcgagtag ctctcttcta ccaatatcgt cattgctgtt tttcttgttt
     3661 ttcactttcg ttctttggat tgtgcttcac ccctcagtat cccttccctt tgtttttatt
     3721 tcctgcgaac attaacaact gcatgaattt tgtacttctc cttttaatcc acgttccggt
     3781 aaggcatcat ccaaattttt ttattcgacc tcgttaagtc atatattttt tcccaaaaat
     3841 acataaaaca ataatgcagc cttcttttca atatttacaa cttttcaatt tatattgtct
     3901 tttgttattt atactcttat atattaaatt tattccgtta ctaaataccc ttttgctata
     3961 caaatatcat caaagagaag tactgaaagc ttacttttta tgcgctgggt aatttttccg
     4021 gaaacaataa cgaaatcatc gtcgagcaat tttgctcgta cttcagaaac tactgcgtaa
     4081 acatttgagg tcggggggat ccactagttc tagagcggcc gccaccgcgg tggagctcca
     4141 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     4201 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4261 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4321 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4381 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4441 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4501 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4561 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4621 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4681 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4741 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4801 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4861 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4921 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4981 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5041 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5101 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5161 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5221 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5281 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5341 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5401 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5461 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5521 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5581 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5641 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5701 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5761 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5821 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5881 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5941 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6001 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6061 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6121 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6181 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6241 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6301 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgaacgaagc atctgtgctt
     6361 cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
     6421 tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt
     6481 gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc
     6541 tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa
     6601 tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag
     6661 catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt
     6721 ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca
     6781 taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat
     6841 tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt
     6901 gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc
     6961 ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga
     7021 ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa
     7081 acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt
     7141 aggttatata gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt
     7201 gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt
     7261 ttgaaagtgc gtcttcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt
     7321 tctagagaat aggaacttcg gaataggaac ttcaaagcgt ttccgaaaac gagcgcttcc
     7381 gaaaatgcaa cgcgagctgc gcacatacag ctcactgttc acgtcgcacc tatatctgcg
     7441 tgttgcctgt atatatatat acatgagaag aacggcatag tgcgtgttta tgcttaaatg
     7501 cgtacttata tgcgtctatt tatgtaggat gaaaggtagt ctagtacctc ctgtgatatt
     7561 atcccattcc atgcggggta tcgtatgctt ccttcagcac taccctttag ctgttctata
     7621 tgctgccact cctcaattgg attagtctca tccttcaatg ctatcatttc ctttgatatt
     7681 ggatcatact aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     7741 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     7801 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     7861 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     7921 gtactgagag tgcaccatac caccttttca attcatcatt ttttttttat tctttttttt
     7981 gatttcggtt tccttgaaat ttttttgatt cggtaatctc cgaacagaag gaagaacgaa
     8041 ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa gaaacatgaa
     8101 attgcccagt attcttaacc caactgcaca gaacaaaaac ctgcaggaaa cgaagataaa
     8161 tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct gttgctgcca
     8221 agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta
     8281 ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt ttactaaaaa
     8341 cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg ctaaaggcat
     8401 tatccgccaa gtacaatttt ttactcttcg aagacagaaa atttgctgac attggtaata
     8461 cagtcaaatt gcagtactct gcgggtgtat aca
//