LOCUS       sequence-393485-        5669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NeoR nls-scarlet marked rapid RMCE integration vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5669)
  AUTHORS   Nonet ML
  TITLE     Rapid generation of C. elegans single-copy transgenes combining RMCE
            and drug selection.
  JOURNAL   Genetics. 2023 Apr 20:iyad072. doi: 10.1093/genetics/iyad072.
  PUBMED    37079426
REFERENCE   2  (bases 1 to 5669)
  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: "Genetics. 
            2023 Apr 20:iyad072. doi: 10.1093/genetics/iyad072."
COMMENT     Sequence Label: pNeoR2
FEATURES             Location/Qualifiers
     source          1..5669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(101..118)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(272..860)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(352..371)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1031..1822,1823..1891))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1654..1673
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1892..1996)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2022..2477)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2159..2180)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2371..2390
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    complement(2489..2522)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(2592..2625)
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     protein_bind    2646..2679
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             2692..2712
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(4512..5312)
                     /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="GNMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4623..4642)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5233..5252
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gatcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
       61 gcggggagag gcggtttgcg tattgggcga attccgcttc ctcgctcact gactcgctgc
      121 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
      181 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
      241 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
      301 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
      361 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
      421 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
      481 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaacccccca
      541 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
      601 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
      661 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
      721 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
      781 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
      841 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
      901 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
      961 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     1021 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     1081 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     1141 catctggccc cagtgctgca atgataccgc gagatccacg ctcaccggct ccagatttat
     1201 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     1261 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     1321 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     1381 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     1441 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     1501 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     1561 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     1621 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     1681 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     1741 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     1801 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     1861 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     1921 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     1981 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     2041 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2101 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     2161 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2221 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     2281 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2341 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2401 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2461 gccgctacag ggcgcgtcgt cggagctcat aacttcgtat agcatacatt atacgaagtt
     2521 atgcggccgc tctagaacta gtggcgaaga gcctgcaggt aaccacgcgt tgcatgctct
     2581 tcggtacccg ggaagttcct atactatttg aagaatagga acttctgcag gaattcgata
     2641 tcggtgaagt tcctattctc tagaaagtat aggaacttca aaatgggatc cccaaagaag
     2701 aagcgtaagg tgtcgatggt ttccaaggga gaggccgtca tcaaggagtt catgcgtttc
     2761 aaggtccaca tggagggatc catgaacgga cacgagttcg agatcgaggg agagggagag
     2821 ggacgtccat acgagggaac ccaaaccgcc aagctcaagg tcaccaaggt aagtttaaac
     2881 atatatatac taactaaccc tgattattta aattttcagg gaggaccact cccattctcc
     2941 tgggacatcc tctccccaca attcatgtac ggatcccgtg ccttcaccaa gcacccagcc
     3001 gacatcccag actactacaa gcaatccttc ccagagggat tcaagtggga gcgtgtcatg
     3061 aacttcgagg acggaggagc cgtcaccgtc acccaagaca cctccctcga ggacggaacc
     3121 ctcatctaca aggtaagttt aaacagttcg gtactaacta accatacata tttaaatttt
     3181 caggtcaagc tccgtggaac caacttccca ccagacggac cagtcatgca aaagaagacc
     3241 atgggatggg aggcctccac cgagcgtctc tacccagagg acggagtcct caagggagac
     3301 atcaagatgg ccctccgtct caaggacgga ggacgttacc tcgccgactt caaggtaagt
     3361 ttaaacatga ttttactaac taactaatct gatttaaatt ttcagaccac ctacaaggcc
     3421 aagaagccag tccaaatgcc aggagcctac aacgtcgacc gtaagctcga catcacctcc
     3481 cacaacgagg actacaccgt cgtcgagcaa tacgagcgtt ccgagggacg tcactccacc
     3541 ggaggaatgg acgagctcta caaggcctcc cgtcgtcgta aggccaaccc aaccaagctc
     3601 tccgagaacg ccaagaagct cgccaagtag atcaactgct ctaattccat ctcgtttctc
     3661 tcttttcttt caatctcttt acccgtttct cttcccctac gtcccccaaa cctaccactg
     3721 cacagtaaca catacacatt caacgtgtca caataaaagc ccatcaacac cttctctgtt
     3781 gttttttttc ttcaaatgca tttttttcac tttgtttaat aatttttacg catcggattt
     3841 gtaggatagt ggaaatttgt ccttcaatgc ttcaattgat tcgacacttc tcaatatttt
     3901 tgatcaataa accattgatt ttctatttag aagaagatca ctgtaaataa caccaaaaac
     3961 atcacagcaa aaaaactccc actcaattat attactgttt ttattgaaac tagaataaat
     4021 ttgaaaatct tcaaagcata cacaattaaa aaaaacgaaa aaagaaaaag tcccgttgat
     4081 tctcaaactt aaatttttaa acttgtctga accatgttgt aagtttgtaa aagtttccca
     4141 gttgtcctcg aacaggtcta aaaattgcgg tcataaactg aaacgtaaca tatgataagg
     4201 tattttgtgt gcggtttttt ctatgatgtt tcaaaaaaat ttatcagaag taaaaaacat
     4261 aagaagtgta cacagaacat tgtgtagatt ttgggataga attattatta atggggggag
     4321 ggagcacaat ttttttcgac tttttattac gaattaattc tatttttgaa tctacacaat
     4381 ttcattgtta gaggtgactt aaaagaagct aaaaaacaaa gaaattaaga gaagtttttt
     4441 gataataaca aaaatagggg gtgggagcac agggagaaag agcatgtagg gatgttgaag
     4501 agtaattgga ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
     4561 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcctca
     4621 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
     4681 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
     4741 ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt
     4801 tcggctggtg cgagcccctg atgttcttcg tccagatcat cctgatcgac aagaccggct
     4861 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
     4921 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
     4981 ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
     5041 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
     5101 cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg caccggacag gtcggtcttg
     5161 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     5221 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     5281 gcgtgcaatc catcttgttc aatcatatta ccttaaaatt caaaaattaa tttcagatca
     5341 gtcataacca agaaaacaaa ataaacgtta ggacagaaaa ttcaaattaa ttaaaaatta
     5401 tgctgaataa gaaaattaca tgcaaagcct tattttcctc atagaaacag ttgaataaca
     5461 tattttttaa ttgaaaacat agcaaaattc gtggtaaaat ttgtcaaaac cttggaaaaa
     5521 atcgaaaaaa gaacatttcc cgatgcattt tcctatttct cgttgccgac aacgtataaa
     5581 cgttgaatac tcagtcaacg aatttggctg ctggcttgga ctctcgcacg gaagagacgc
     5641 gtgtgtagat ttacgacgaa agcaaaaat
//
