pRS426_SOD2_GFP_LplA

LOCUS       pRS426_SOD2_GFP_LplA              8578 bp DNA     circular SYN 13-MAY-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8578)
  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..8578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(775..879)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      906..2248
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(1869..1916)
                     /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        2510..2725
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             2726..3529
                     /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"
     rep_origin      complement(3660..4115)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     4256..4272
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4282..4300
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4326..4342
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4583..4870
                     /gene="S. cerevisiae TDH3"
                     /label=TDH3 promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     CDS             4875..4952
                     /label=sceSOD2
     misc_feature    4959..5672
                     /label=GFP
     misc_feature    5673..5708
                     /label=linker
     CDS             5709..6752
                     /label=LplA
     CDS             6732..6749
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      6763..7234
                     /label=SSD1 terminator
     primer_bind     complement(7239..7255)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(7292..7310)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7331..7347)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7355..7371
                     /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(7379..7409)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7424..7445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7733..8321)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(8492..8578,1..774))
                     /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"
ORIGIN
        1 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
       61 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
      121 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
      181 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
      241 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
      301 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
      361 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
      421 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
      481 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
      541 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
      601 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
      661 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
      721 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
      781 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
      841 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
      901 acctgaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa
      961 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc
     1021 tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc
     1081 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag
     1141 agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg
     1201 agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta
     1261 taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac
     1321 tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat
     1381 tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt atattctata
     1441 ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg
     1501 gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt
     1561 ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt
     1621 ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt
     1681 caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa
     1741 agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag tagctcgtta
     1801 cagtccggtg cgtttttggt tttttgaaag tgcgtcttca gagcgctttt ggttttcaaa
     1861 agcgctctga agttcctata ctttctagag aataggaact tcggaatagg aacttcaaag
     1921 cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc tgcgcacata cagctcactg
     1981 ttcacgtcgc acctatatct gcgtgttgcc tgtatatata tatacatgag aagaacggca
     2041 tagtgcgtgt ttatgcttaa atgcgtactt atatgcgtct atttatgtag gatgaaaggt
     2101 agtctagtac ctcctgtgat attatcccat tccatgcggg gtatcgtatg cttccttcag
     2161 cactaccctt tagctgttct atatgctgcc actcctcaat tggattagtc tcatccttca
     2221 atgctatcat ttcctttgat attggatcat actaagaaac cattattatc atgacattaa
     2281 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     2341 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     2401 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     2461 actatgcggc atcagagcag attgtactga gagtgcacca taccaccttt tcaattcatc
     2521 attttttttt tattcttttt tttgatttcg gtttccttga aatttttttg attcggtaat
     2581 ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat
     2641 atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa
     2701 aacctgcagg aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac
     2761 tcatcctagt cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt
     2821 gtgtgcttca ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg
     2881 tcccaaaatt tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg
     2941 cacagttaag ccgctaaagg cattatccgc caagtacaat tttttactct tcgaagacag
     3001 aaaatttgct gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat
     3061 agcagaatgg gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg
     3121 tttgaagcag gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga
     3181 attgtcatgc aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc
     3241 gaagagcgac aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga
     3301 tgaaggttac gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagacgc
     3361 attgggtcaa cagtatagaa ccgtggatga tgtggtctct acaggatctg acattattat
     3421 tgttggaaga ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag
     3481 aaaagcaggc tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt
     3541 ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt
     3601 attaccctat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga
     3661 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3721 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3781 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3841 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3901 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3961 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     4021 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     4081 acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac
     4141 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     4201 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     4261 acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg taccgggccc
     4321 cccctcgagg tcgacggtat cgataagctt gatatcgaat tcctgcagcc cacagtttat
     4381 tcctggcatc cactaaatat aatggagccc gctttttaag ctggcatcca gaaaaaaaag
     4441 aatcccagca ccaaaatatt gttttcttca ccaaccatca gttcataggt ccattctctt
     4501 agcgcaacta cagagaacag gggcacaaac aggcaaaaaa cgggcacaac ctcaatggag
     4561 tgatgcaacc tgcctggagt aaatgatgac acaaggcaat tgacccacgc atgtatctat
     4621 ctcattttct tacaccttct attaccttct gctctctctg atttggaaaa agctgaaaaa
     4681 aaaggttgaa accagttccc tgaaattatt cccctacttg actaataagt atataaagac
     4741 gttaggtatt gattgtaatt ctgtaaatct atttcttaaa cttcttaaat tctactttta
     4801 tagttagtct tttttttagt tttaaaacac caagaactta gtttcgaata aacacacata
     4861 aacaaacaaa gatgatgttc gctaagaccg ctgctgctaa tttgaccaag aagggtggtt
     4921 tgtctttgtt gtctaccacc gcaagaagaa ccggtaccgt ttctaaaggt gaagaattgt
     4981 ttactggtgt tgttccaatt ttggttgaat tggatggtga tgttaatggt cataaatttt
     5041 ctgtttccgg tgaaggtgaa ggtgatgcta cttatggtaa attgactttg aagtttattt
     5101 gtaccaccgg taaattgcct gttccatgac caactttggt tactacttta acttatggtg
     5161 ttcaatgttt ctctagatat ccagatcata tgaagcaaca tgatttcttc aaatctgcta
     5221 tgccagaagg ttatgttcaa gaaagaacta ttttcttcaa ggatgatggt aattacaaga
     5281 ccagagctga agttaagttt gaaggtgata ctttagttaa cagaattgaa ttgaagggta
     5341 ttgatttcaa ggaagatggt aatattttgg gtcataagtt ggaatataac tacaattctc
     5401 ataacgttta cattatggct gataaacaaa agaacggtat taaagttaac ttcaagatta
     5461 gacataacat tgaagatggt tcagttcaat tggctgatca ttatcaacaa aacaccccaa
     5521 ttggtgatgg tccagttttg ttgccagata atcattactt gtctactcaa tcaaagttgt
     5581 caaaagatcc aaatgaaaag agagatcata tggttttatt ggaatttgtt accgctgctg
     5641 gtattacttt gggtatggat gaattgtaca aatctagagc tgatggttct ggttccggtt
     5701 ctgagctcat gggttcaact ttgagattgt tgatttctga ttcttacgat ccatggttta
     5761 acttggctgt tgaagaatgt attttcagac aaatgccagc tactcaaaga gttttgtttt
     5821 tgtggagaaa tgctgatact gttgttattg gtagagcaca aaatccttgg aaggaatgta
     5881 atactagaag aatggaagaa gataacgtta gattggctag aagatcaagt ggtggtggtg
     5941 ctgttttcca tgatttaggt aatacatgtt tcactttcat ggctggtaaa ccagaatacg
     6001 ataaaacaat ttcaacctct attgttttga acgctttgaa tgctttgggt gttagtgctg
     6061 aagctagtgg tagaaatgat ttggttgtta aaactgttga aggtgataga aaagtttctg
     6121 gttctgctta tagagaaact aaggatagag gttttcatca tggtactttg ttgttgaatg
     6181 ctgatttgtc cggattggct aattatttga atcctgataa gaagaagttg gctgctaaag
     6241 gtattacttc agttagatca agagttacta acttaactga attgttgcct ggtattactc
     6301 atgaacaagt ttgtgaagct attactgaag catttttcgc tcattatggt gaaagagttg
     6361 aagctgaaat tatttctcca aataagaccc cagatttgcc taattttgct gaaacattcg
     6421 ctagacaatc ttcttgggaa tggaattttg gtcaagctcc agcttttagt catttgttgg
     6481 atgaaagatt tacctggggt ggtgttgaat tgcattttga tgttgaaaag ggtcatatta
     6541 caagagcaca agttttcact gattctttga atccagctcc attggaagct ctagctggta
     6601 gattgcaagg ttgtttgtat agagctgata tgttgcaaca agaatgtgaa gcattattgg
     6661 ttgattttcc agaacaagaa aaggaattga gagaattgag tgcttggatg gctggtgcag
     6721 ttagagaatt ccatcatcat catcatcatt aactcgagta gctctcttct accaatatcg
     6781 tcattgctgt ttttcttgtt tttcactttc gttctttgga ttgtgcttca cccctcagta
     6841 tcacttccct ttgtttttat ttcctgcgaa cattaacaac tgcatgaatt ttgtacttct
     6901 ccttttaatc cacgttccgg taaggcatca tccaaatttt tttatttgac ctcgttaagt
     6961 catatatttt ttcccaaaaa tacataaaac aataatgcag ccttcttttc aatatttaca
     7021 acttttcaat ttatattgtc ttttgttatt tatactctta tatattaaat ttattccgtt
     7081 actaaatacc cttttgctgt acaaatatca tcaaagagaa gtactgaaag cttacttttt
     7141 atgcgctggg taatttttcc ggaaacaata acgaaatcat cgtcgagcaa ttttgctcgt
     7201 acttcagaaa ctactgcgta aacatttgag gtcgggggga tccactagtt ctagagcggc
     7261 cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     7321 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     7381 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     7441 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     7501 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     7561 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     7621 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     7681 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     7741 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     7801 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     7861 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     7921 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     7981 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     8041 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     8101 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     8161 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     8221 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     8281 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     8341 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     8401 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     8461 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     8521 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagat
//