LOCUS       sequence-329697-        5883 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SiMPl plasmid pair (pSiMPls) for use with 
            spectinomycin/streptomycin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5883)
  AUTHORS   Palanisamy N, Ballestin JB, Di Ventura B
  TITLE     Expanding the SiMPl Plasmid Toolbox for Use with 
            Spectinomycin/Streptomycin.
  JOURNAL   ACS Omega. 2021 May 21;6(22):14148-14153. doi: 
            10.1021/acsomega.1c00649. eCollection 2021 Jun 8.
  PUBMED    34124437
REFERENCE   2  (bases 1 to 5883)
  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"; doi: 
            "10.1021/acsomega.1c00649"; journalName: "ACS Omega"; date: 
            "2021-05-21- 21"; volume: "6"; issue: "22"; pages: "14148-14153"
COMMENT     Sequence Label: pSiMPls_N + pSiMPls_C
FEATURES             Location/Qualifiers
     source          1..5883
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     178..197
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     437..458
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(545..829)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     complement(603..622)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             856..1734
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     complement(1914..1933)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      2009..2554
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     2455..2474
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2671..2688
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3080..3182
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      complement(4230..4685)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4367..4388)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4579..4598
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        complement(5143..5234)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(5253..5280)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5372..5458)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5611..5628
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(join(5667..5883,1..497,498..500,501..503))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(5670..5883,1..497,498..500,501..503))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5712..5733)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg
       61 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
      121 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
      181 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
      241 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
      301 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
      361 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
      421 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
      481 agctcctcgc ccttgctcac catcctggcc ttgagctcga attcgctagc ccaaaaaaac
      541 gggtatggag aaacagtaga gagttgcgat aaaaagcgtc aggtaggatc cgctaatctt
      601 atggataaaa atgctatggc atagcaaagt gtgacgccgt gcaaataatc aatgtggact
      661 tttctgccgt gattatagac acttttgtta cgcgtttttg tcatggcttt ggtcccgctt
      721 tgttacagaa tgcttttaat aagcggggtt accggtttgg ttagcgagaa gagccagtaa
      781 aagacgcagt gacggcaatg tctgatgcaa tatggacaat tggtttcttc tctgaatggc
      841 gggagtatga aaagtatggc tgaagcgcaa aatgatcccc tgctgccggg atactcgttt
      901 aatgcccatc tggtggcggg tttaacgccg attgaggcca acggttatct cgattttttt
      961 atcgaccgac cgctgggaat gaaaggttat attctcaatc tcaccattcg cggtcagggg
     1021 gtggtgaaaa atcagggacg agaatttgtt tgccgaccgg gtgatatttt gctgttcccg
     1081 ccaggagaga ttcatcacta cggtcgtcat ccggaggctc gcgaatggta tcaccagtgg
     1141 gtttactttc gtccgcgcgc ctactggcat gaatggctta actggccgtc aatatttgcc
     1201 aatacggggt tctttcgccc ggatgaagcg caccagccgc atttcagcga cctgtttggg
     1261 caaatcatta acgccgggca aggggaaggg cgctattcgg agctgctggc gataaatctg
     1321 cttgagcaat tgttactgcg gcgcatggaa gcgattaacg agtcgctcca tccaccgatg
     1381 gataatcggg tacgcgaggc ttgtcagtac atcagcgatc acctggcaga cagcaatttt
     1441 gatatcgcca gcgtcgcaca gcatgtttgc ttgtcgccgt cgcgtctgtc acatcttttc
     1501 cgccagcagt tagggattag cgtcttaagc tggcgcgagg accaacgtat cagccaggcg
     1561 aagctgcttt tgagcaccac ccggatgcct atcgccaccg tcggtcgcaa tgttggtttt
     1621 gacgatcaac tctatttctc gcgggtattt aaaaaatgca ccggggccag cccgagcgag
     1681 ttccgtgccg gttgtgaaga aaaagtgaat gatgtagccg tcaagttgtc ataattggta
     1741 acgaatcaga caattgacgg cttgacggag tagcataggg tttgcagaat ccctgcttcg
     1801 tccatttgac aggcacatta tgcatcgatg ataagctgtc aaacatgagc agatcctcta
     1861 cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat
     1921 cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga gcctagaaat
     1981 attttatctg attaataaga tgatcttctt gagatcgttt tggtctgcgc gtaatctctt
     2041 gctctgaaaa cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc tgagctacca
     2101 actctttgaa ccgaggtaac tggcttggag gagcgcagtc accaaaactt gtcctttcag
     2161 tttagcctta accggcgcat gacttcaaga ctaactcctc taaatcaatt accagtggct
     2221 gctgccagtg gtgcttttgc atgtctttcc gggttggact caagacgata gttaccggat
     2281 aaggcgcagc ggtcggactg aacggggggt tcgtgcatac agtccagctt ggagcgaact
     2341 gcctacccgg aactgagtgt caggcgtgga atgagacaaa cgcggccata acagcggaat
     2401 gacaccggta aaccgaaagg caggaacagg agagcgcacg agggagccgc cagggggaaa
     2461 cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc
     2521 gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg ccctctcact
     2581 tccctgttaa gtatcttcct ggcatcttcc aggaaatctc cgccccgttc gtaagccatt
     2641 tccgctcgcc gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga agcggaatat
     2701 atcctgtatc acatattctg ctgacgcacc ggtgcagcct tttttctcct gccacatgaa
     2761 gcacttcact gacaccctca tcagtgccaa catagtaagc cagtatacac tccgctagcg
     2821 ctgatgtccg gcggtgcttt tgccgttacg caccaccccg tcagtagctg aacaggaggg
     2881 acagctgata gaaacagaag ccactggagc acctcaaaaa caccatcata cactaaatca
     2941 gtaagttggc agcatcaccc gacgcacttt gcgccgaata aatacctgtg acggaagatc
     3001 acttcgcaga ataaataaat cctggtgtcc ctgttgatac cgggaagccc tgggccaact
     3061 tttggcgaaa atgagacgtt gatcggcacg taagaggttc caactttcac cataatgaaa
     3121 taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta
     3181 aaatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca
     3241 tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg
     3301 gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg
     3361 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttgc ttggatctga
     3421 aaacccaggt tcagaccccg cagggtatga aggaaatttc caacatccag gtcggtgatc
     3481 tggtactgag caacacgggt tacaacgaag ttctgaacgt cttcccgaaa tctaaaaaaa
     3541 agtcttacaa aatcaccctg gaagatggca aggaaatcat ctgttccgaa gaacacctgt
     3601 ttccgacgca gactggtgaa atgaacatct ccggtggtct gaaagaaggt atgtgtctgt
     3661 atgttaaaga ataatttttt taaggcagtt attggtgccc ttaaacgcct ggttgctacg
     3721 cctgaataag tgataataag cggatgaatg gcagaaattc gaaagcaaat tcgacccggt
     3781 cgtcggttca gggcagggtc gttaaatagc cgcttatgtc tattgctggt ttaccggttt
     3841 attgactacc ggaagcagtg tgaccgtgtg cttctcaaat gcctgactgg taacaggatt
     3901 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3961 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4021 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4081 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4141 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4201 tcaaaaagga tcttcaccta gatcctttta aattgtaaac gttaatattt tgttaaaatt
     4261 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     4321 cccttataaa tcaaaagaat agcccgagat agggttgagt gttgttccag tttggaacaa
     4381 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     4441 cgatggccca ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa
     4501 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     4561 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     4621 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4681 cgcgtaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4741 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4801 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4861 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4921 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4981 gccgggaagc tagagtaagt agttcgccag ttaatagttt gctgcaccca actgatcttc
     5041 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5101 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5161 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5221 gaaaaataaa caaaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct
     5281 gcttaatttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt
     5341 gcttcgcaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg
     5401 acaaacaaca gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat
     5461 gcctggcagt tccctactct cgcatgggga gaccccacac taccatcggc gctacggcgt
     5521 ttcacttctg agttcggcat ggggtcaggt gggaccaccg cgctactgcc gccaggcaaa
     5581 ttctgtttta tcagaccgct tctgcgttct gatttaatct gtatcaggct gaaaatcttc
     5641 tctcatccgc caaaacagcc aagcttttac ttgtacagct cgtccatgcc gagagtgatc
     5701 ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg
     5761 ctcagggcgg actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg
     5821 atgggggtgt tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg
     5881 cgg
//
