LOCUS       sequence-348603-        4747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid with p15A origin for constitutive mCherry expression in E. 
            coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4747)
  TITLE     Plasmids with p15A origin for constitutive fluorescent protein 
            expression in E. coli
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4747)
  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            "Unpublished"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: pmr101A-cPr-mCherry (KmR)
FEATURES             Location/Qualifiers
     source          1..4747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     350..369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     632..651
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             773..1483
                     /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(918..936)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1186..1205
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1551..1569)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1565..1612
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2214..3029)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2214..3029)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2937..2956
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3021..3040
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3490..3507)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3624..4169)
                     /direction=LEFT
                     /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     complement(3704..3723)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaattgtaaa cgtcaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcctc aattaggaag cagcccagta
      481 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
      541 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
      601 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
      661 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgaagatc
      721 taatacttga catatcactg tgattcacat ataatatgcg aaggagctcg agatggtgag
      781 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
      841 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
      901 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
      961 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
     1021 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     1081 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
     1141 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
     1201 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
     1261 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
     1321 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
     1381 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
     1441 gggccgccac tccaccggcg gcatggacga gctgtacaag tagtaatcga gctcaagctt
     1501 ggatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     1561 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     1621 aggaactata tccggatatc cacaggacgg gtgtggtcgc catgatggtg accaccattt
     1681 gccccatatc accgtcggta ctgatcccgt cgtcaataaa ccgaaccgct acaccctgag
     1741 catcaaactc ttttatcagt tggatcatgt cggcggtgtc gcggccaaga cggtcgagct
     1801 tcttcaccag aatgacatca ccttcctcca ccttcatcct cagcaaatcc agcccttccc
     1861 gatctgttga actgccggat gccttgtcgg taaagatgcg gttagctttt acccctgcat
     1921 ctttgagcgc tgaggtctgc ctcgtgaaga aggtgttgct gactcatacc aggcctgaat
     1981 cgccccatca tccagccaga aagtgaggga gccacggttg atgagagctt tgttgtaggt
     2041 ggaccagttg gtgattttga acttttgctt tgccacggaa cggtctgcgt tgtcgggaag
     2101 atgcgtgatc tgatccttca actcagcaaa agttcgattt attcaacaaa gccgccgtcc
     2161 cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat taaccaattc tgattagaaa
     2221 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     2281 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     2341 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     2401 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     2461 ggtgagaatg gcaagagctt atgcatttct ttccagactt gttcaacagg ccagccatta
     2521 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     2581 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     2641 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     2701 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
     2761 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     2821 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     2881 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     2941 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggccttgag
     3001 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     3061 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
     3121 tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagatc
     3181 acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac
     3241 tggtccacct acaacaaagc tctcatcaac cgtggctccc tcactttctg gctggatgat
     3301 ggggcgattc aggcctggta tgagtcagca acaccttctt cacgaggcag acctcagcgc
     3361 tagcggagtg tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc
     3421 atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata
     3481 ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat
     3541 ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta acagggaagt
     3601 gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa
     3661 atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3721 ccccctggcg gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt
     3781 ccgctgttat ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt
     3841 tcgctccaag ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc
     3901 cggtaactat cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc
     3961 cactggtaat tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg
     4021 aaaggacaag ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg
     4081 tagctcagag aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag
     4141 agattacgcg cagaccaaaa cgatctcaag aagatcatct tattaagggg tctgacgctc
     4201 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4261 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4321 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4381 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4441 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4501 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4561 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4621 atagtttgcg caacgttgtt gccattgctg cagggttgaa ggctctcaag ggcatcggtc
     4681 gacgctctcc cttatgcgac tcctgcgaaa agccccaaaa acaggaagat tgtataagca
     4741 aatattt
//
