LOCUS       sequence-480267-        6032 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 6032)
  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 6032)
  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: pSS2.1-mNeongreen
FEATURES             Location/Qualifiers
     source          1..6032
                     /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(4313..5107)
                     /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(4424..4443)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5034..5053
                     /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 ctagatccag gtacgacatt atacttgtta
     2041 tgttttacaa atgaattggt aacaacactt gaatttatca aaaaatatcc atcaattatt
     2101 tctgatattc ttatttatgg ttgttctgga gcaattggac aattgtttat tttccataca
     2161 ttagaaaaat atggcagttt aatcttaata actatcacac ttacaagaaa aatgctaacg
     2221 ctattattaa gtcttatttg gtttaatcat aagttaacaa ttggccagtg gataggtgta
     2281 gtattggtat tttatggagc aagtttggaa gcatatataa aacatacaca attagctaaa
     2341 aaaaaagtat aaaacattta tacgaaatct catacgtaaa tttataaaat ttataataaa
     2401 catataaata atataaaaat gaaacaatat tctaaaaaca aatctataac aaaaaacata
     2461 tattttaatt acgctacata attaatatat aatagcaata ccatcaatat actgcttcct
     2521 taattacgag tatacttaag aggtatcaat tcctcaatag ttgttttaac aagtttacca
     2581 ttcatctcaa taagaacaaa acaatctggc gcataatctg atataagctc tcgacacata
     2641 ccacatggag aaacaacacg aatagaccta tcaacttcat cagaataagg atgtcgaact
     2701 gcaacaatag tatcaaaatc cttttgtcca ttagatactg cacttccaat agctattgcc
     2761 tctgcacaaa cagtaacacg tcctatataa gcttcaatgt gaacagctga tataatctct
     2821 ccagtctttg ttctaattgc tgctccaaca tggtgtttat tatcctcata aagcattgta
     2881 atcttttctg tagcaacttc tactaattct aaatcctgct gagatatatt aaatgtcttc
     2941 atcttttttt taatatattt ttttcaaata tctaaatatt attttatgtt aaaatgaatt
     3001 tttgaacgtc gatattcaaa aaatatcatg actgttttta tttttttgac ctaatttttt
     3061 cagattaaga ataaatattt aacattaaga tttttagatt atacaataaa gacgtcattt
     3121 cttatgacaa ctagtccaaa taactacata attataaaaa ttattagaaa aatctacgtt
     3181 tctaaaacca attcaatatt atatatatat atatatagcc ttcataatta ttctagaaaa
     3241 caatttattt taatactatt cttttaaagc tacaatattc aatattatca tctatatcat
     3301 tattattaca tatctttaat atttttcaga aaaataatta taaaaataaa acgtgccatc
     3361 atatttttta ttgaaattaa atctatgaag ttttaaatca atctaaagta tatatgagta
     3421 accggttctg acagcaccca gaaacgctgg tcaaagtaaa agatgctgaa gaccagttgg
     3481 gtgcgcgggt gggttacatc gaactggatc tcaatagcgg taagatcctt gagagttttc
     3541 gccccgaaga acgtttccca atgatgagca cctttaaagt tctgctgtgt ggcgcggtgt
     3601 tatcccgtat tgacgccggt caagagcagc tcggtcgccg cattcactat tcgcagaatg
     3661 acttggttga atatagtccg gtcacagaaa agcatctgac ggatggcatg acagtcagag
     3721 aactgtgctc agctgccata accatgagcg ataacactgc ggccaactta cttctgacca
     3781 cgatcggagg accgaaagaa ctgaccgcct tcttgcacaa catgggggat catgtgactc
     3841 gcttagatcg ttgggaaccg gagctgaatg aagcgatacc gaacgacgag cgtgacacca
     3901 cgatgcctgt ggcaatggca acaaccttgc gcaaactatt aactggcgaa cttctgactc
     3961 tcgcttcccg gcaacaactg attgactgga tggaagcgga taaagttgca ggacctctgc
     4021 tgcgctcggc ccttccggct ggctggttta ttgcggataa atctggcgcc ggggagcgtg
     4081 gctctcgcgg tattattgca gcgctggggc cggatggtaa acccagtcgt atcgtagtga
     4141 tctacacgac cggcagccag gcaaccatgg atgaacgaaa taggcagatc gcggagattg
     4201 gcgcctcact gattaagcat tggtaaagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4261 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca attcagaaga
     4321 actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa
     4381 gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca
     4441 acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa
     4501 agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat
     4561 cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct
     4621 gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc
     4681 gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca
     4741 gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg tgggatgaca
     4801 ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa
     4861 cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct
     4921 cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc
     4981 cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt
     5041 catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt
     5101 caatcatgcg aaacgatcct catcctgtct cttgatcaga tcttgatccc ctgcgccatc
     5161 agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca accttaccag
     5221 agggcgcccc agctggcaat tccggttcgc ttgctgtcca tagtagttct agaatttttg
     5281 ttaaaatttt atttcaagag ttgatgggag ttcttggtat acgcgaatta gatgaacgtt
     5341 tgaaagatcc acttttacaa gagactcttt ctaatatgtt tcctcgggat aatccaaaaa
     5401 atactagatt tgcaataaat tattttactt ctattggatt aggattgata actgtttcat
     5461 tgcgtgagca tttggaaact atcaaggtaa agctttgaaa atatagtatt aactctaatt
     5521 tctttagaat actgttacag taaatcgaag tctttctaaa tcttcatcat cttcaattat
     5581 atcttcctat actacatcat attcatctga atctttatca tatcccaatg atacttcaga
     5641 gtcagaatca ttgaatcgtt caagatctcg tcatcggtca cctgctcgtc atagaatgag
     5701 atcccctcga gataatcgtt caaggccacg ttcatgttca ttaaatcgtt ctagattgca
     5761 atcttctgag tcggaatcat tgaatcgttc gagatctcgt catcggtcac ctccgcccag
     5821 tctagctatc gccatgtaag cccactgcaa gctacctgct ttctctttgc gcttgcgttt
     5881 tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga
     5941 ctggctttct acgtgttccg cttcctttag cagcatgaat cagaaatttg gggataatgg
     6001 gaatttttta ctatatacaa aggaccagct ca
//
