LOCUS       sequence-480269-        4137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mNeongreen under control of Msg promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4137)
  AUTHORS   Sayson SG, Ashbaugh A, Bauer LC, Smulian AG
  TITLE     Extracellular vesicle-mediated delivery of genetic material for 
            transformation and CRISPR/Cas9-based gene editing in Pneumocystis 
            murina.
  JOURNAL   mBio. 2025 Sep 23:e0182525. doi: 10.1128/mbio.01825-25.
  PUBMED    40985725
REFERENCE   2  (bases 1 to 4137)
  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: "mBio. 2025 
            Sep 23:e0182525. doi: 10.1128/mbio.01825-25."
COMMENT     Sequence Label: pSS1-mNeongreen
FEATURES             Location/Qualifiers
     source          1..4137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..25
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7..25
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1096..1115)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1097..1115)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(1327..1915)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1407..1426)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2960..3754)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3071..3090)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3681..3700
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gctagcaatt aaccctcact aaagggagac tctacgtcaa ttgcacaaat ttgagaaaag
       61 agacggatat caagaaaaat atatttttca aacgctaaaa caaataacgg gtaaagcaaa
      121 tacacagtcc ggaaataacc cctttaatac gaaaataata attaacattc tcataaactc
      181 ccgatctgac ctaaggtcaa aaaaatggtt tcaaagggcg aagaggataa catggcctct
      241 ctcccagcga cacatgagtt acacatattt ggctctatta acggtgttga ttttgacatg
      301 gtcggtcaag gaacgggtaa tccaaatgat ggttatgaag agttaaacct gaaaagcact
      361 aagggagatc ttcaattctc tccttggatt ttagtccctc atatcgggta tggctttcat
      421 cagtatcttc cctaccctga cggtatgagt ccttttcagg cagccatggt agatggatcc
      481 ggataccaag tacatcgaac aatgcaattt gaagatggtg cctcccttac tgttaattat
      541 agatatacct acgaaggaag ccacattaaa ggagaagctc aagttaaggg gactggtttt
      601 cctgctgacg gtcctgtgat gactaattct ttgaccgctg ctgactggtg taggtcgaag
      661 aaaacttacc ccaacgataa aacgataatt agtaccttta aatggtcata tactactgga
      721 aatggtaaac gctaccgttc aactgctaga accacatata cttttgcaaa accaatggct
      781 gctaattatt tgaagaatca gccgatgtat gtgttccgta aaacggaatt gaagcattct
      841 aagacagaac taaatttcaa agaatggcaa aaagcattta cagatgttat gggcatggat
      901 gagttatata aataaggtga gtgacactac gacgataagg agaatgtact ggattttgat
      961 aagcgttgtt atcttgatta tttacttcaa tttatttttt gtttttatat ttgtttttaa
     1021 tattgtattc tagtctttaa actgtttatt taaatagtat tgccttaaag caatacgtag
     1081 aatgctactc accctcccta tagtgagtcg tattaggcca caatcgtgaa tgtgtttgcg
     1141 aatttagagg tatgtatttt atcagattat tttactataa cattcataat caaattatat
     1201 ttttaaaata ttacttcaag tgtatatcat gtataattac acggcataca aaaaaaaaaa
     1261 ctcgtcaata aataaataga atttatttta ttaactacag gaatgtgatg gccgcgttgc
     1321 tggcgttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1381 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1441 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1501 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1561 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1621 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1681 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1741 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1801 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1861 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1921 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1981 tttggtcatg agattatcaa agaagatctt ctagatcctt ttaaattaaa aatgaagttt
     2041 taaatcaatc taaagtatat atgagtaacc ggttctgaca gcacccagaa acgctggtca
     2101 aagtaaaaga tgctgaagac cagttgggtg cgcgggtggg ttacatcgaa ctggatctca
     2161 atagcggtaa gatccttgag agttttcgcc ccgaagaacg tttcccaatg atgagcacct
     2221 ttaaagttct gctgtgtggc gcggtgttat cccgtattga cgccggtcaa gagcagctcg
     2281 gtcgccgcat tcactattcg cagaatgact tggttgaata tagtccggtc acagaaaagc
     2341 atctgacgga tggcatgaca gtcagagaac tgtgctcagc tgccataacc atgagcgata
     2401 acactgcggc caacttactt ctgaccacga tcggaggacc gaaagaactg accgccttct
     2461 tgcacaacat gggggatcat gtgactcgct tagatcgttg ggaaccggag ctgaatgaag
     2521 cgataccgaa cgacgagcgt gacaccacga tgcctgtggc aatggcaaca accttgcgca
     2581 aactattaac tggcgaactt ctgactctcg cttcccggca acaactgatt gactggatgg
     2641 aagcggataa agttgcagga cctctgctgc gctcggccct tccggctggc tggtttattg
     2701 cggataaatc tggcgccggg gagcgtggct ctcgcggtat tattgcagcg ctggggccgg
     2761 atggtaaacc cagtcgtatc gtagtgatct acacgaccgg cagccaggca accatggatg
     2821 aacgaaatag gcagatcgcg gagattggcg cctcactgat taagcattgg taaagcaaaa
     2881 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     2941 atactcttcc tttttcaatt cagaagaact cgtcaagaag gcgatagaag gcgatgcgct
     3001 gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa
     3061 gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca
     3121 gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc
     3181 aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg
     3241 cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa
     3301 gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg
     3361 ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt
     3421 tctcggcagg agcaaggtgg gatgacagga gatcctgccc cggcacttcg cccaatagca
     3481 gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg
     3541 tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca ccggacaggt
     3601 cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag
     3661 agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg
     3721 gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt
     3781 gatcagatct tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta
     3841 ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc ggttcgcttg
     3901 ctgtccataa aaccgcccag tctagctatc gccatgtaag cccactgcaa gctacctgct
     3961 ttctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg
     4021 gtcagcaccg tttctgcgga ctggctttct acgtgttccg cttcctttag cagcatgaat
     4081 cagaaatttg gggataatgg gaatttttta ctatatacaa aggaccagct cagctag
//
