LOCUS       sequence-486017-        4742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Neo resistant attB phiC31 recombination-mediation insertion (RMI) 
            vector for C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4742)
  AUTHORS   Beck AA, Nonet ML
  TITLE     A single component landing site for efficient transgenesis using 
            recombination-mediated insertion.
  JOURNAL   MicroPubl Biol. 2025 Sep 2;2025:10.17912/micropub.biology.001784. 
            doi: 10.17912/micropub.biology.001784. eCollection 2025.
  PUBMED    40970081
REFERENCE   2  (bases 1 to 4742)
  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: "MicroPubl 
            Biol."; date: "2025-09-2"; volume: 
            "2025:10.17912/micropub.biology.001784. doi"; pages: " 
            10.17912/micropub.biology.001784. eCollection 2025"
COMMENT     Sequence Label: pAttBF3Neo
FEATURES             Location/Qualifiers
     source          1..4742
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(21..40)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      834..1422
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1323..1342
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1576..1593
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1709..1730
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1745..1750,1751..1768,1769..1775)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1783..1799
                     /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     1788..1810
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1807..1823
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1807..1823
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1842..1862
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1844..1862
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    1879..1912
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    1976..2009
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             complement(2385..3185)
                     /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(2496..2515)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3106..3125
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     protein_bind    3563..3596
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(3701..3734)
                     /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)"
     primer_bind     complement(3772..3791)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3773..3791)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3801..3818)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3801..3817)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3810..3832)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3959..4414
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4046..4065)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4256..4277
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4440..4544
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4545..4613,4614..4742,1..663)
                     /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 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
       61 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
      121 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
      181 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
      241 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
      301 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
      361 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
      421 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
      481 tggtttattg ctgataaatc tggagccggt gagcgtggat ctcgcggtat cattgcagca
      541 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
      601 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
      661 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
      721 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
      781 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
      841 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
      901 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
      961 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     1021 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     1081 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     1141 cggtcgggct gaatgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     1201 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     1261 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     1321 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     1381 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     1441 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     1501 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     1561 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aattcgccca atacgcaaac
     1621 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     1681 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     1741 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     1801 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
     1861 gggaaagccg cggtgcgggt gccagggcgt gcccttgggc tccccgggcg cgtactccac
     1921 ctcacccatc tggtccatca tgatgaacgg gtcgaggtgg cggtagttga tacaaataac
     1981 ttcgtataaa gtatcctata cgaagttatc ccagttgtcc tcgaacaggt ctaaaaattg
     2041 cggtcataaa ctgaaacgta acatatgata aggtattttg tgtgcggttt tttctatgat
     2101 gtttcaaaaa aatttatcag aagtaaaaaa cataagaagt gtacacagaa cattgtgtag
     2161 attttgggat agaattatta ttaatggggg gagggagcac aatttttttc gactttttat
     2221 tacgaattaa ttctattttt gaatctacac aatttcattg ttagaggtga cttaaaagaa
     2281 gctaaaaaac aaagaaatta agagaagttt tttgataata acaaaaatag ggggtgggag
     2341 cacagggaga aagagcatgt agggatgttg aagagtaatt ggactcagaa gaactcgtca
     2401 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
     2461 aagcggtcag cccattcgcc gccaagctcc tcagcaatat cacgggtagc caacgctatg
     2521 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
     2581 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
     2641 tcgggcatgc gcgccttgag cctggcgaac agttcggctg gtgcgagccc ctgatgttct
     2701 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
     2761 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
     2821 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
     2881 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
     2941 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc
     3001 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
     3061 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
     3121 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcata
     3181 ttaccttaaa attcaaaaat taatttcaga tcagtcataa ccaagaaaac aaaataaacg
     3241 ttaggacaga aaattcaaat taattaaaaa ttatgctgaa taagaaaatt acatgcaaag
     3301 ccttattttc ctcatagaaa cagttgaata acatattttt taattgaaaa catagcaaaa
     3361 ttcgtggtaa aatttgtcaa aaccttggaa aaaatcgaaa aaagaacatt tcccgatgca
     3421 ttttcctatt tctcgttgcc gacaacgtat aaacgttgaa tactcagtca acgaatttgg
     3481 ctgctggctt ggactctcgc acggaagaga cgcgtgtgta gatttacgac gaaagcaaaa
     3541 atgatcccgc tttccagtcg ggataacttc gtataaagta tcctatacga agttataagc
     3601 tggagctcca ccgcggtggc ggccgctcta gatggcgaag agcccatgga tccactagta
     3661 acatatgtgc aggcctgctc ttcggtaagc ttatcgatac gaagttccta tactatttga
     3721 agaataggaa cttccgtcga cccgttaacc gggggcccgg tacccaattc gccctatagt
     3781 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     3841 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     3901 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     3961 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4021 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4081 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4141 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4201 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4261 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4321 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4381 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     4441 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4501 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4561 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4621 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4681 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4741 tg
//
