LOCUS       sequence-486011-        6222 bp    DNA     circular SYN 15-DEC-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 6222)
  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 6222)
  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: "bioRxiv 
            2024.03.01.583017"
COMMENT     Sequence Label: pAttBFGNeo2
FEATURES             Location/Qualifiers
     source          1..6222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     621..640
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(859..963)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(989..1444)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1126..1147)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1338..1357
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    complement(1550..1583)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1616..1632
                     /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     1621..1643
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1640..1656
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1640..1656
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1675..1695
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1677..1695
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1843..1862)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1844..1862)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1872..1889)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1872..1888)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1945..1966
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2082..2099)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2193..2262
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     protein_bind    2304..2337
                     /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             2350..2370
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(4233..5033)
                     /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(4344..4363)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4954..4973
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      complement(5461..6049)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5541..5560)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6220..6222,1..789,790..858))
                     /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"
ORIGIN
        1 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
       61 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
      121 tgctgcaatg ataccgcgag atccacgctc accggctcca gatttatcag caataaacca
      181 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
      241 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
      301 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
      361 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
      421 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
      481 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
      541 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
      601 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
      661 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
      721 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
      781 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
      841 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
      901 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
      961 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     1021 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     1081 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     1141 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     1201 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     1261 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     1321 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     1381 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     1441 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     1501 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaaa taacttcgta
     1561 tagcatacat tatacgaagt tatgttgggt aacgccaggg ttttcttgtg tggaattgtg
     1621 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     1681 aaccctcact aaagggaaca aaagctggag ctccaccgcg gtggcggccg ctctagatgg
     1741 cgaagagccc atggatccac tagtaacata tgtgcaggcc tgctcttcgg taagcttatc
     1801 gataccgtcg acccgttaac cgggggcccg gtacccaatt cgccctatag tgagtcgtat
     1861 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact ggcatacgag ccggaagcat
     1921 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     1981 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     2041 cgcggggaga ggcggtttgc gtattgggcg aattccgctt cctcgctcac tgactcgctg
     2101 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2161 tccacagaat caggggataa cgcaggaaag aacatgaagc cgcggtgcgg gtgccagggc
     2221 gtgcccttgg gctccccggg cgcgtactcc acctcaccca tctggtccat catgatgaac
     2281 gggtcgaggt ggcggtagtt gatgaagttc ctattctcta gaaagtatag gaacttcaaa
     2341 atgggatccc caaagaagaa gcgtaaggtg tcgatggtgt cgaagggaga agaggataac
     2401 atggcttcac tcccagctac acacgaactc cacatcttcg gatcgatcaa cggagtggat
     2461 ttcgatatgg tcggacaagg taagtttaaa catatatata ctaactaacc ctgattattt
     2521 aaattttcag gaactggaaa cccaaacgat ggatacgagg aactcaacct caagtcgaca
     2581 aagggagatc tgcaattctc gccatggatt ctcgtgccac acatcggata cggattccac
     2641 caatacctcc catacccagg taagtttaaa ctgagttcta ctaactaacg agtaatattt
     2701 aaattttcag atggaatgtc accattccaa gctgccatgg tggatggatc gggataccaa
     2761 gttcaccgaa caatgcaatt cgaggatgga gcctcgctca cagtgaacta ccgatacaca
     2821 tacgagggat cgcacatcaa ggtaagttta aacagttcgg tactaactaa ccatacatat
     2881 ttaaattttc agggagaggc tcaagttaag ggaacaggat tcccagctga tggaccagtg
     2941 atgacaaact cactcacagc tgctgattgg tgccgatcga aaaagacata cccaaatgat
     3001 aagacaatca tctcgacatt caagtggtcg tacactactg gaaacggaaa gcgataccga
     3061 tcgacagccc gaacaacata cacattcgct aagccaatgg ccgccaacta cctcaaggta
     3121 agtttaaaca tgattttact aactaactaa tctgatttaa attttcagaa tcaaccaatg
     3181 tacgtgttcc gaaagacaga actcaagcac tcaaagacag agctgaactt caaagagtgg
     3241 caaaaggcct tcacagatgt gatgggaatg gatgaactct acaaggcctc ccgtcgtcgt
     3301 aaggccaacc caaccaagct ctccgagaac gccaagaagc tcgccaagta gatcaactgc
     3361 tctaattcca tctcgtttct ctcttttctt tcaatctctt tacccgtttc tcttccccta
     3421 cgtcccccaa acctaccact gcacagtaac acatacacat tcaacgtgtc acaataaaag
     3481 cccatcaaca ccttctctgt tgtttttttt cttcaaatgc atttttttca ctttgtttaa
     3541 taatttttac gcatcggatt tgtaggatag tggaaatttg tccttcaatg cttcaattga
     3601 ttcgacactt ctcaatattt ttgatcaata aaccattgat tttctattta gaagaagatc
     3661 actgtaaata acaccaaaaa catcacagca aaaaaactcc cactcaatta tattactgtt
     3721 tttattgaaa ctagaataaa tttgaaaatc ttcaaagcat acacaattaa aaaaaacgaa
     3781 aaaagaaaaa gtcccgttga ttctcaaact taaattttta aacttgtctg aaccatgttg
     3841 taagtttgta aaagtttccc agttgtcctc gaacaggtct aaaaattgcg gtcataaact
     3901 gaaacgtaac atatgataag gtattttgtg tgcggttttt tctatgatgt ttcaaaaaaa
     3961 tttatcagaa gtaaaaaaca taagaagtgt acacagaaca ttgtgtagat tttgggatag
     4021 aattattatt aatgggggga gggagcacaa tttttttcga ctttttatta cgaattaatt
     4081 ctatttttga atctacacaa tttcattgtt agaggtgact taaaagaagc taaaaaacaa
     4141 agaaattaag agaagttttt tgataataac aaaaataggg ggtgggagca cagggagaaa
     4201 gagcatgtag ggatgttgaa gagtaattgg actcagaaga actcgtcaag aaggcgatag
     4261 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
     4321 cattcgccgc caagctcctc agcaatatca cgggtagcca acgctatgtc ctgatagcgg
     4381 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
     4441 atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgcgc
     4501 gccttgagcc tggcgaacag ttcggctggt gcgagcccct gatgttcttc gtccagatca
     4561 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
     4621 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
     4681 atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact
     4741 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
     4801 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg
     4861 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     4921 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     4981 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatatt accttaaaat
     5041 tcaaaaatta atttcagatc agtcataacc aagaaaacaa aataaacgtt aggacagaaa
     5101 attcaaatta attaaaaatt atgctgaata agaaaattac atgcaaagcc ttattttcct
     5161 catagaaaca gttgaataac atatttttta attgaaaaca tagcaaaatt cgtggtaaaa
     5221 tttgtcaaaa ccttggaaaa aatcgaaaaa agaacatttc ccgatgcatt ttcctatttc
     5281 tcgttgccga caacgtataa acgttgaata ctcagtcaac gaatttggct gctggcttgg
     5341 actctcgcac ggaagagacg cgtgtgtaga tttacgacga aagcaaaaat gatcccgctt
     5401 tccgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5461 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5521 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5581 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5641 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5701 caagctgggc tgtgtgcacg aaccccccat tcagcccgac cgctgcgcct tatccggtaa
     5761 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5821 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5881 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5941 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6001 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6061 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6121 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6181 atcaatctaa agtatatatg agtaaacttg gtctgacagt ta
//
