LOCUS       sequence-11532-e        4634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4634)
  AUTHORS   Frokjaer-Jensen C, Davis MW, Hollopeter G, Taylor J, Harris TW, Nix 
            P, Lofgren R, Prestgard-Duke M, Bastiani M, Moerman DG, Jorgensen EM
  TITLE     Targeted gene deletions in C. elegans using transposon excision.
  JOURNAL   Nat Methods. 2010 Jun;7(6):451-3. doi: 10.1038/nmeth.1454. Epub 2010
            Apr 25.
  PUBMED    20418868
REFERENCE   2  (bases 1 to 4634)
  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"; doi: 
            "10.1038/nmeth.1454"; journalName: "Nat Methods"; date: "2010-06"; 
            volume: "7"; issue: "6"; pages: "451-3"
COMMENT     Sequence Label: pRL8
FEATURES             Location/Qualifiers
     source          1..4634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..18
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2..18
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    34..133
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(279..290)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(2224..2323)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2340..2359)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2341..2359)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2364..2380)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2364..2380)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2482..2501)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2493..3302
                     /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(2560..2579)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3449..4037
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3938..3957
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4191..4208
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4367..4394)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4486..4572)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4621..4634,1..9)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tgtaaaacga cggccagtct taagctcggg ccccaaataa tgattttatt ttgactgata
       61 gtgacctgtt cgttgcaaca aattgatgag caatgctttt ttataatgcc aactttgtac
      121 aaaaaagcag gcttgagcca atttatccaa gtccttgtaa aaagtattcg aaaattgtta
      181 acggataaaa tgtttattat aatatcaaaa acaatttgtc agttgaccac tttttgatat
      241 atttgacaga aaccgggatg aattggctca aagttagggc gcccttctat tacagggttt
      301 ctgataacaa accggttatt aactcccaac aagggatgat ttcaattcat catgctcaaa
      361 ttgacccaaa ttaagttaca tgacaaattc atcgcccttt tcaactcttc ttggctcatc
      421 atctgttatt ctgttcctat tctgtctgca ccctataccc ctttgcatac tctctcgtca
      481 ttcctttctt ggatagtgtc ttcttctccc cagctctgct acttctatga cttgcgcccg
      541 ctgcttttcc gctcgttcct ctcctctctg acgtcttctt ctgcttcttt tctccttcct
      601 ttttcccttc tcaatcttcc tctttttccc atttctgtca atcattcgaa gaagaagaag
      661 aagaagaccc tcattcattt catttttttt tctgtcggtg tgtgctgctc ggttaagagt
      721 gagctctcta ttccacgtct tcttcttttt cttcttcgat tcgaatcaat cactccacca
      781 aaacgcattc gtttttggga ttcaccccgc ggttcgcaat aggtttcttt ttcaaattat
      841 tagcgttata aatagaaaaa tggtggagtt tcaataaaaa tgataattta caaagtgtat
      901 tttgattaca tgtactcaaa aaggttgaaa tttttcatac cagttttccg gaaatccatc
      961 tgatatcatt atcgtatttt tctttttaaa aatgttttca aaaaaaaaca aaatatagct
     1021 ggttatttgg caccctctaa ttaccatttt cctgtcacac cacactcttt tccttcttcc
     1081 acttctttcc gttttcagcc gcttccaacc aaaccgatat gaaagccgag caacaacgat
     1141 cgattccacc cggctcggcg accttcccgt cgcaggtgag actcagaaaa ctagagaaac
     1201 cgctcaacta actcttgata tccgatttca ttcttttctc ttttcttttt gttgaacttt
     1261 tccatatttc cagatgccac ggccaccacc aagcaccgaa caaggaatca caacggaatc
     1321 ggagcttgcg aagaaagctc aaatcactcc gaacgacgtt ttagcacttc cgggaatcac
     1381 tcaaggtatc ttgctccctt tggatttcag tttaaaacat aaatttagga ttcttatgct
     1441 ccccatctgc gaacatctat aacatcgagt tcaccaagtt ccaaatccgt gatctggaca
     1501 ctgagcaagt gcttttcgag atcgccaaac cggagaacga tcaggagaat gatgagtcgc
     1561 cacaggagtc ggcaagatac gtgcgttata gatttgctcc aaactttttg aaactcaaaa
     1621 cggtcggagc aactgtggaa ttcaaagtag gagacatccc aatccatcat ttccgaatga
     1681 tcgaacgtca cttcttcaaa gatcgccttc tgaagtgttt tgactttgaa ttcggattct
     1741 gtattccgaa ttcacgaaac aactgtgaac atatctatga gttccctcaa ctctctcaac
     1801 aactcagtga gtcattattc taaaaagtac aattcaaaag actaatctct ttttcagtgg
     1861 acgacatgat caacaatcca aacgagactc gttctgacag cttctatttc gtcgataaca
     1921 aactcgtcat gcacaacaaa gccgactact catatgatgc ataaatattt aatacaaaaa
     1981 tgttctggat aattattctg tcgaatagaa aaaaaactcc aaatgtgatt aaattccaat
     2041 aattcctgtc tagtttgttc cttccccttc cccttctcat gttcaatgca ttcctaagct
     2101 tttcagttcc ccccttgttt ctatattttt tcgctgtcct gtcacactcg ctaaaaacac
     2161 taatcacacg gaaatctgtt ttcaataaaa actccaactt taactcattt tcaatttcaa
     2221 ctgacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt
     2281 gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc
     2341 cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc
     2401 aaaatctctg atgttacatt gcacaagata aaataatatc atcatgaaca ataaaactgt
     2461 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga
     2521 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
     2581 atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt
     2641 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
     2701 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
     2761 atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag
     2821 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
     2881 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg
     2941 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
     3001 gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt
     3061 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
     3121 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
     3181 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
     3241 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
     3301 aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg
     3361 gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     3421 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3481 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3541 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3601 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3661 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3721 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3781 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3841 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3901 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3961 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4021 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4081 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4141 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4201 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     4261 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     4321 caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt
     4381 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
     4441 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag
     4501 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt
     4561 tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggatgtttt
     4621 cccagtcacg acgt
//
