LOCUS       sequence-348605-        4744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid with p15A origin for constitutive mNeonGreen expression in 
            E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4744)
  TITLE     Plasmids with p15A origin for constitutive fluorescent protein 
            expression in E. coli
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4744)
  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            "Unpublished"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-mNeonGreen (KmR)
FEATURES             Location/Qualifiers
     source          1..4744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(499..546)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     542..560
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     primer_bind     complement(1457..1476)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      1652..2107
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1739..1758)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1949..1970
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2686..3231
                     /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     3132..3151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3348..3365
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3815..3834)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3826..4641
                     /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             3826..4641
                     /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     complement(3899..3918)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gatcacgcat cttcccgaca acgcagaccg ttccgtggca aagcaaaagt tcaaaatcac
       61 caactggtcc acctacaaca aagctctcat caaccgtggc tccctcactt tctggctgga
      121 tgatggggcg attcaggcct ggtatgagtc agcaacacct tcttcacgag gcagacctca
      181 gcgctcaaag atgcaggggt aaaagctaac cgcatcttta ccgacaaggc atccggcagt
      241 tcaacagatc gggaagggct ggatttgctg aggatgaagg tggaggaagg tgatgtcatt
      301 ctggtgaaga agctcgaccg tcttggccgc gacaccgccg acatgatcca actgataaaa
      361 gagtttgatg ctcagggtgt agcggttcgg tttattgacg acgggatcag taccgacggt
      421 gatatggggc aaatggtggt caccatcatg gcgaccacac ccgtcctgtg gatatccgga
      481 tatagttcct cctttcagca aaaaacccct caagacccgt ttagaggccc caaggggtta
      541 tgctagttat tgctcagcgg tggcagcagc caactcagct tcctttcggg ctttgttagc
      601 agccggatcc aagcttgagc tcgattactt atataattca tccattccca tgacatcggt
      661 gaaggctttt tgccattcct taaagttcaa ctcggtctta ctatgtttta gttcagtctt
      721 cctaaacacg tacatgggtt gatttttaag ataattagcg gccattggtt tagcgaatgt
      781 atacgtagta cgggcagtgg atctgtatct tttaccattg ccagtggtat aagaccactt
      841 aaaagttgaa attattgttt tatcgttagg atatgtcttt ttagacctac accagtcagc
      901 agctgtcagg ctattagtca taacgggacc atcagccgga aaaccagtac ccttaacttg
      961 ggcttctccc ttaatatggg aaccctcgta cgtgtacctg tagtttactg ttaaggaagc
     1021 gccgtcttca aactgcatgg ttctatgaac ttggtagcca gaaccgtcta ccatagcggc
     1081 ttgaaatgga ctcattccat cagggtaagg taaatattga tgaaagccgt acccaatatg
     1141 gggcacaaga atccaggggg aaaattgcaa atcgcctttt gttgatttaa gattaagttc
     1201 ctcgtatcca tcgtttggat taccagtgcc ttgtccaacc atatcaaagt caacaccatt
     1261 aatacttccg aaaatatgta attcgtgagt tgctggcaag gatgccatgt tgtcctcctc
     1321 cccttttgag accatccatg gctccttcgc atattatatg tgaatcacag tgatatgtca
     1381 agtattagat cttcgatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca
     1441 ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac
     1501 ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga
     1561 ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc
     1621 aacctactac tgggctgctt cctaattgag gacgcgccct gtagcggcgc attaagcgcg
     1681 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     1741 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     1801 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     1861 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     1921 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     1981 aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     2041 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attgacgttt
     2101 acaatttaaa tatttgctta tacaatcttc ctgtttttgg ggcttttcgc aggagtcgca
     2161 taagggagag cgtcgaccga tgcccttgag agccttcaac cctgcagcaa tggcaacaac
     2221 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     2281 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     2341 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     2401 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     2461 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     2521 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     2581 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     2641 gttttcgttc cactgagcgt cagacccctt aataagatga tcttcttgag atcgttttgg
     2701 tctgcgcgta atctcttgct ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa
     2761 ggttctctga gctaccaact ctttgaaccg aggtaactgg cttggaggag cgcagtcacc
     2821 aaaacttgtc ctttcagttt agccttaacc ggcgcatgac ttcaagacta actcctctaa
     2881 atcaattacc agtggctgct gccagtggtg cttttgcatg tctttccggg ttggactcaa
     2941 gacgatagtt accggataag gcgcagcggt cggactgaac ggggggttcg tgcatacagt
     3001 ccagcttgga gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg agacaaacgc
     3061 ggccataaca gcggaatgac accggtaaac cgaaaggcag gaacaggaga gcgcacgagg
     3121 gagccgccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccaccactga
     3181 tttgagcgtc agatttcgtg atgcttgtca ggggggcgga gcctatggaa aaacggcttt
     3241 gccgcggccc tctcacttcc ctgttaagta tcttcctggc atcttccagg aaatctccgc
     3301 cccgttcgta agccatttcc gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga
     3361 gcgaggaagc ggaatatatc ctgtatcaca tattctgctg acgcaccggt gcagcctttt
     3421 ttctcctgcc acatgaagca cttcactgac accctcatca gtgccaacat agtaagccag
     3481 tatacactcc gctagcgctg aggtctgcct cgtgaagaag gtgttgctga ctcataccag
     3541 gcctgaatcg ccccatcatc cagccagaaa gtgagggagc cacggttgat gagagctttg
     3601 ttgtaggtgg accagttggt gattttgaac ttttgctttg ccacggaacg gtctgcgttg
     3661 tcgggaagat gcgtgatctg atccttcaac tcagcaaaag ttcgatttat tcaacaaagc
     3721 cacgttgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga
     3781 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
     3841 cgggaaacgt cttgctcaag gccgcgatta aattccaaca tggatgctga tttatatggg
     3901 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg
     3961 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     4021 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     4081 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca
     4141 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     4201 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     4261 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     4321 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataagctc
     4381 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     4441 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     4501 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     4561 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     4621 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt
     4681 acgctgactt gacgggacgg cggctttgtt gaataaatcg aacttttgct gagttgaagg
     4741 atca
//
