LOCUS       sequence-486006-        6235 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  attB Neo Resistant mNeonGreen marked plasmid for RMI targeting an 
            attP site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6235)
  AUTHORS   Nonet ML
  TITLE     Creation and manipulation of bipartite expression transgenes in C. 
            elegans using phiC31 recombinase.
  JOURNAL   bioRxiv 2024.03.01.583017
REFERENCE   2  (bases 1 to 6235)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2024.03.01.583017"
COMMENT     Sequence Label: pAttBFGNeo
FEATURES             Location/Qualifiers
     source          1..6235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(723..1523)
                     /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(834..853)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1444..1463
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     protein_bind    complement(1907..1940)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(1998..2586)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2078..2097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2757..3548,3549..3617))
                     /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     3380..3399
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3618..3722)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3748..4203)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3885..3906)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4097..4116
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4341..4357
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4346..4368
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4365..4381
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4365..4381
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4400..4420
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4402..4420
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4568..4587)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4569..4587)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4597..4614)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4597..4613)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4670..4691
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4807..4824)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4918..4987
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     protein_bind    5029..5062
                     /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             5075..5095
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 cttcaaatgc atttttttca ctttgtttaa taatttttac gcatcggatt tgtaggatag
       61 tggaaatttg tccttcaatg cttcaattga ttcgacactt ctcaatattt ttgatcaata
      121 aaccattgat tttctattta gaagaagatc actgtaaata acaccaaaaa catcacagca
      181 aaaaaactcc cactcaatta tattactgtt tttattgaaa ctagaataaa tttgaaaatc
      241 ttcaaagcat acacaattaa aaaaaacgaa aaaagaaaaa gtcccgttga ttctcaaact
      301 taaattttta aacttgtctg aaccatgttg taagtttgta aaagtttccc agttgtcctc
      361 gaacaggtct aaaaattgcg gtcataaact gaaacgtaac atatgataag gtattttgtg
      421 tgcggttttt tctatgatgt ttcaaaaaaa tttatcagaa gtaaaaaaca taagaagtgt
      481 acacagaaca ttgtgtagat tttgggatag aattattatt aatgggggga gggagcacaa
      541 tttttttcga ctttttatta cgaattaatt ctatttttga atctacacaa tttcattgtt
      601 agaggtgact taaaagaagc taaaaaacaa agaaattaag agaagttttt tgataataac
      661 aaaaataggg ggtgggagca cagggagaaa gagcatgtag ggatgttgaa gagtaattgg
      721 actcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
      781 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcctc agcaatatca
      841 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg
      901 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
      961 acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggt
     1021 gcgagcccct gatgttcttc gtccagatca tcctgatcga caagaccggc ttccatccga
     1081 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     1141 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     1201 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     1261 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     1321 cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     1381 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     1441 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     1501 ccatcttgtt caatcatatt accttaaaat tcaaaaatta atttcagatc agtcataacc
     1561 aagaaaacaa aataaacgtt aggacagaaa attcaaatta attaaaaatt atgctgaata
     1621 agaaaattac atgcaaagcc ttattttcct catagaaaca gttgaataac atatttttta
     1681 attgaaaaca tagcaaaatt cgtggtaaaa tttgtcaaaa ccttggaaaa aatcgaaaaa
     1741 agaacatttc ccgatgcatt ttcctatttc tcgttgccga caacgtataa acgttgaata
     1801 ctcagtcaac gaatttggct gctggcttgg actctcgcac ggaagagacg cgtgtgtaga
     1861 tttacgacga aagcaaaaat gatcccgctt tccagtcggg aaacctataa cttcgtatag
     1921 catacattat acgaagttat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     1981 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2041 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2101 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2161 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2221 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccattca gcccgaccgc
     2281 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2341 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2401 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2461 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2521 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2581 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2641 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     2701 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2761 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2821 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     2881 gctgcaatga taccgcgaga tccacgctca ccggctccag atttatcagc aataaaccag
     2941 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3001 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3061 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3121 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3181 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3241 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3301 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3361 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3421 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3481 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3541 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3601 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     3661 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     3721 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     3781 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     3841 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     3901 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     3961 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     4021 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     4081 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     4141 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     4201 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     4261 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     4321 cagggttttc ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     4381 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctcca
     4441 ccgcggtggc ggccgctcta gatggcgaag agcccatgga tccactagta acatatgtgc
     4501 aggcctgctc ttcggtaagc ttatcgatac cgtcgacccg ttaaccgggg gcccggtacc
     4561 caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg
     4621 tgactggcat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4681 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4741 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgaattc
     4801 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4861 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4921 gaagccgcgg tgcgggtgcc agggcgtgcc cttgggctcc ccgggcgcgt actccacctc
     4981 acccatctgg tccatcatga tgaacgggtc gaggtggcgg tagttgatga agttcctatt
     5041 ctctagaaag tataggaact tcaaaatggg atccccaaag aagaagcgta aggtgtcgat
     5101 ggtgtcgaag ggagaagagg ataacatggc ttcactccca gctacacacg aactccacat
     5161 cttcggatcg atcaacggag tggatttcga tatggtcgga caaggtaagt ttaaacatat
     5221 atatactaac taaccctgat tatttaaatt ttcaggaact ggaaacccaa acgatggata
     5281 cgaggaactc aacctcaagt cgacaaaggg agatctgcaa ttctcgccat ggattctcgt
     5341 gccacacatc ggatacggat tccaccaata cctcccatac ccaggtaagt ttaaactgag
     5401 ttctactaac taacgagtaa tatttaaatt ttcagatgga atgtcaccat tccaagctgc
     5461 catggtggat ggatcgggat accaagttca ccgaacaatg caattcgagg atggagcctc
     5521 gctcacagtg aactaccgat acacatacga gggatcgcac atcaaggtaa gtttaaacag
     5581 ttcggtacta actaaccata catatttaaa ttttcaggga gaggctcaag ttaagggaac
     5641 aggattccca gctgatggac cagtgatgac aaactcactc acagctgctg attggtgccg
     5701 atcgaaaaag acatacccaa atgataagac aatcatctcg acattcaagt ggtcgtacac
     5761 tactggaaac ggaaagcgat accgatcgac agcccgaaca acatacacat tcgctaagcc
     5821 aatggccgcc aactacctca aggtaagttt aaacatgatt ttactaacta actaatctga
     5881 tttaaatttt cagaatcaac caatgtacgt gttccgaaag acagaactca agcactcaaa
     5941 gacagagctg aacttcaaag agtggcaaaa ggccttcaca gatgtgatgg gaatggatga
     6001 actctacaag gcctcccgtc gtcgtaaggc caacccaacc aagctctccg agaacgccaa
     6061 gaagctcgcc aagtagatca actgctctaa ttccatctcg tttctctctt ttctttcaat
     6121 ctctttaccc gtttctcttc ccctacgtcc cccaaaccta ccactgcaca gtaacacata
     6181 cacattcaac gtgtcacaat aaaagcccat caacaccttc tctgttgttt ttttt
//
