LOCUS       sequence-485477-        4700 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  attB Neo Resistant plasmid for RMI targeting an attP site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4700)
  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 4700)
  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: pAttBFNeo2
FEATURES             Location/Qualifiers
     source          1..4700
                     /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             complement(2711..3511)
                     /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(2822..2841)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3432..3451
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      complement(3939..4527)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4019..4038)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4698..4700,1..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 gaacttccag
     2341 ttgtcctcga acaggtctaa aaattgcggt cataaactga aacgtaacat atgataaggt
     2401 attttgtgtg cggttttttc tatgatgttt caaaaaaatt tatcagaagt aaaaaacata
     2461 agaagtgtac acagaacatt gtgtagattt tgggatagaa ttattattaa tggggggagg
     2521 gagcacaatt tttttcgact ttttattacg aattaattct atttttgaat ctacacaatt
     2581 tcattgttag aggtgactta aaagaagcta aaaaacaaag aaattaagag aagttttttg
     2641 ataataacaa aaataggggg tgggagcaca gggagaaaga gcatgtaggg atgttgaaga
     2701 gtaattggac tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     2761 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcctcag
     2821 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     2881 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     2941 catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt
     3001 cggctggtgc gagcccctga tgttcttcgt ccagatcatc ctgatcgaca agaccggctt
     3061 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     3121 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     3181 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     3241 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     3301 acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga
     3361 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     3421 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     3481 cgtgcaatcc atcttgttca atcatattac cttaaaattc aaaaattaat ttcagatcag
     3541 tcataaccaa gaaaacaaaa taaacgttag gacagaaaat tcaaattaat taaaaattat
     3601 gctgaataag aaaattacat gcaaagcctt attttcctca tagaaacagt tgaataacat
     3661 attttttaat tgaaaacata gcaaaattcg tggtaaaatt tgtcaaaacc ttggaaaaaa
     3721 tcgaaaaaag aacatttccc gatgcatttt cctatttctc gttgccgaca acgtataaac
     3781 gttgaatact cagtcaacga atttggctgc tggcttggac tctcgcacgg aagagacgcg
     3841 tgtgtagatt tacgacgaaa gcaaaaatga tcccgctttc cgtgagcaaa aggccagcaa
     3901 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3961 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4021 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4081 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4141 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4201 ccccccattc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4261 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4321 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4381 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4441 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4501 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4561 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4621 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4681 taaacttggt ctgacagtta
//
