LOCUS       sequence-42179-e        4895 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4895)
  AUTHORS   Frokjaer-Jensen C, Davis MW, Ailion M, Jorgensen EM
  TITLE     Improved Mos1-mediated transgenesis in C. elegans.
  JOURNAL   Nat Methods. 2012 Jan 30;9(2):117-8. doi: 10.1038/nmeth.1865.
  PUBMED    22290181
REFERENCE   2  (bases 1 to 4895)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nmeth"; 
            journalName: "Nat Methods"; date: "2012-01-30- 30"; volume: "9"; 
            issue: "2"; pages: "117-8"
COMMENT     Sequence Label: pCFJ594 - pENTR[1-2](NeoR)
FEATURES             Location/Qualifiers
     source          1..4895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..810
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(68..87)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      957..1545
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1446..1465
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1699..1716
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(1875..1902)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1994..2080)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2129..2151
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2143..2160
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2144..2160
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2176..2275
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3097..3891
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3151..3170)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3761..3780
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4592..4608
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4593..4609
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4627..4726)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4743..4762)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4744..4762)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4767..4783)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4767..4783)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(join(4885..4895,1..9))
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
       61 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
      121 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
      181 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
      241 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
      301 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
      361 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
      421 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
      481 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
      541 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
      601 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
      661 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
      721 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
      781 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
      841 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
      901 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
      961 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1021 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1081 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1141 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1201 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1261 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1321 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1381 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1441 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1501 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1561 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     1621 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     1681 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     1741 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     1801 gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag
     1861 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
     1921 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
     1981 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     2041 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
     2101 ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt
     2161 cttaagctcg ggccccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa
     2221 cacattgatg agcaatgctt ttttataatg ccaactttgt acaaaaaagc aggctcacgt
     2281 gggccggatg ggccatggca ccatttctaa attttagttt aataaaaaaa ttttaaaata
     2341 atacgagttt tgagactaac tttcaataaa aaccaaaaaa ttgcttcaaa aaagagctcg
     2401 cgccgattta cgataaacac agcgcgctac gacactccga aaaccgcgcc gcaccattac
     2461 attctccgcc ccaccgcacg gtttgcggaa ggcgatgaaa ttgctgggcc taaaatttta
     2521 accctttttt tcccgttttt caactgtttt cccgattttt tcgatatttt tttcaggttt
     2581 ttagttagaa aagtatataa aaattgcatt atttgtagtt ttcgctcgta atttgtgata
     2641 gaaaacggcc aatttgccta gatttgtgat tctaccaagt ggaatttttg ggaaaatatc
     2701 aatattccac gatttttatt caaatcataa cgtttcgaaa ttatcagctt aaataaaact
     2761 aaaataaata ataaaatact aaaaaccctg aatttctcgg ttctcggttc agcttgagtt
     2821 cgttctatac acgttatcga tttttgtcat ttcgtttaat catgggtttt gattcatttt
     2881 aaatatgaaa tttgatttat tcctcgtttc gaagttggtt tgtcttgtcc ggccggtaaa
     2941 caaatgttat taaaaaaaat aataaatttc gaaatttttc gtaatgttcc gcaatcgtgt
     3001 ttttccgttt atctcgtctt cccttccttc aaatatacta tttattttct caattcacat
     3061 attttcagcc agccctgctc aacaagtgga ataaaaatga ttgaacaaga tggattgcac
     3121 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3181 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3241 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     3301 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3361 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3421 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3481 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3541 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcaccagcc
     3601 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     3661 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     3721 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     3781 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     3841 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agtccaatta
     3901 ctcttcaaca tccctacatg ctctttctcc ctgtgctccc accccctatt tttgttatta
     3961 tcaaaaaact tctcttaatt tctttgtttt ttagcttctt ttaagccacc tctaacaatg
     4021 aaattgtgta gattcaaaaa tagaattaat tcgtaataaa aagtcgaaaa aaattgtgct
     4081 ccctcccccc attaataata attctatccc aaaatctaca caatgttctg tgtacacttc
     4141 ttatgttttt tacttctgat aaattttttt gaaacatcat agaaaaaacc gcacacaaaa
     4201 taccttatca tatgttacgt ttcagtttat gaccgcaatt tttatttctt cgcacgtctg
     4261 ggcctctcat gacgtcaaat catgctcatc gtgaaaaagt tttggagtat ttttggaatt
     4321 tttcaatcaa gtgaaagttt atgaaattaa ttttcctgct tttgcttttt ggggtttccc
     4381 ctattgtttg tcaagatttc gaggacggcg tttttcttgc taaaatcaca agtattgatg
     4441 agcacgatgc aagaaagatc ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta
     4501 gaagttgata atttgaaagt ggagtagtgt ctatggggtt tttgccttaa atgacagaat
     4561 acattcccaa tataccaaac ataactgttt ctcgaggtcg acggtatcga taagcttgat
     4621 cccgggaccc agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt
     4681 gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata
     4741 tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt
     4801 ctcaaaatct ctgatgttac attgcacaag ataaaataat atcatcatga acaataaaac
     4861 tgtctgctta cataaacagt aatacaaggg gtgtt
//
