LOCUS       sequence-60721-e        6276 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 6276)
  AUTHORS   Frokjaer-Jensen C, Davis MW, Sarov M, Taylor J, Flibotte S, LaBella 
            M, Pozniakovsky A, Moerman DG, Jorgensen EM
  TITLE     Random and targeted transgene insertion in Caenorhabditis elegans 
            using a modified Mos1 transposon.
  JOURNAL   Nat Methods. 2014 May;11(5):529-34. doi: 10.1038/nmeth.2889. Epub 
            2014 Mar 16.
  PUBMED    24820376
REFERENCE   2  (bases 1 to 6276)
  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.2889"; journalName: "Nat Methods"; date: "2014-05"; 
            volume: "11"; issue: "5"; pages: "529-34"
COMMENT     Sequence Label: pCFJ1200
FEATURES             Location/Qualifiers
     source          1..6276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     283..299
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     283..299
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    319..339
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_feature    339..446
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     363..379
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     363..379
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(413..429)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(414..430)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    complement(446..466)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(474..491)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(474..490)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1105..1124)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1703..1723
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     terminator      3694..4017
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(4161..4180)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4280..4302
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4340..4358)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4426..4530
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4531..4599,4600..5391)
                     /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"
     primer_bind     complement(4749..4768)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5562..6150
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6051..6070
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcaccgtacg tctcgaggaa ttctatcagg tgtacaagta tgaaatgtcg tttcgaacat
       61 atagatgtct cgcaaacgta aatatttatc gattgtcata aaactttgac cttgtgaagt
      121 gtcaaccttg actgtcgaac caccatagtt tggcgcgaat tgagcgtcat aattgtttac
      181 tctcagtgca gtcaacatgt cgagtttcgt gccgaataaa gagcaaacgc ggacagtatt
      241 aattttctgt tttcatttga agaaaacagc tgcggccgga tccaggaaac agctatgacc
      301 atgattacgc caagctatca actttgtata gaaaagttga gctccaccgc ggtggcggcc
      361 gctctagaac tagtggatcc cccgggctgc aggaattcga tatcaagctt atcgataccg
      421 tcgacctcga gggggggccc ggtaccaact ttattataca tagttgataa ttcactggcc
      481 gtcgttttac agaggggaga aaatgcggag agaagaaccc agcatcttaa taaaattata
      541 taattttgaa taaaatcaaa ttaaaaataa gattaattga aacacaacaa tagaatgcag
      601 aaaattactc ataagaaata atttattttg aaagaagttg tgtcgattta cgataaaaac
      661 tacacgcaaa cgaggctccg cgtttacacc gtttggcaat ggagcgcgct tgctattaga
      721 ttttaggcca gctcaacgaa agtttaattt atttttcttc tttttttcca tttttttgag
      781 taaaatatcg aattttttgt attaattctg taaattttat aatttgcaac tattttggcg
      841 tattctgctt gtttttatca gcgaattttt atgcatttca gtcaaaagtt tgagcaggaa
      901 aaatgacatt taggactttg taaaattgta ttcttaaatt tgcttgaaaa cctacgtgta
      961 caattgattt tattgaattt ttcttacgtt tcccgttgta tcttattgta tttaactttc
     1021 atcataaaat aatttatttc caggtaaaat atccgacgct ctcgtggatc cccgggattg
     1081 gctcagaaaa aaaaggtacc gaccatgacc gagtacaagc ccacggtgcg cctcgccacc
     1141 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     1201 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     1261 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcggtaagtt taaacatata
     1321 tatactaact aaccctgatt atttaaattt tcagcggacg acggcgccgc ggtggcggtc
     1381 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     1441 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     1501 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgacgtaagt
     1561 ttaaacatga ttttactaac taactaatct gatttaaatt ttcagcacca gggcaagggt
     1621 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     1681 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     1741 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     1801 gcctgatgag ctagccggcc atacaagtaa tccggatgat cgacgccaac gtcgttgaat
     1861 tttcaaattt taaatactga atatttgttt tttttcctat tatttattta ttctctttgt
     1921 gttttttttc ttgctttcta aaaaattaat tcaatccaaa tctaaacatt tttttttctc
     1981 tttccgtctc ccaattcgta ttccgctcct ctcatctgaa cacaatgtgc aagtttattt
     2041 atcttctcgc tttcatttca ttaggacgtg gggggaattg gtggaagggg gaaacacaca
     2101 aaaggatgat ggaaatgaaa taaggacaca caatatgcaa caacattcaa ttcagaaata
     2161 tggaggaagg tttaaaagaa aacataaaaa tatatagagg aggaaggaaa ggtgccggat
     2221 ctgcggctta ctcaccagac ctggccccat ccgattacca cctattcgct tcgatgggac
     2281 acgcactcgc tgagcagcgc ttcgattctt acgaaagtgt gaaaaaatgg ctcgatgaat
     2341 ggttcgccgc aaaagacgat gagttctact ggcgtggaat ccacaaattg cccgagagat
     2401 gggaaaaatg tgtagctagc gacggcaaat actttgaata aatgattttt tctttttcca
     2461 caaaatttaa cgtgtttttt gatttaaaaa aaacgacatt tcatacttgt acacctgata
     2521 tctagatacc taggtgagct ctggtacccc cctggcgtta cccaacttaa tcgccttgca
     2581 gccaacttaa tcaccatgca gccaacttaa tcgccttgca gccaacttaa tcgccttgca
     2641 gccaacttaa tcgccttgca accatgcagg tcgactctag aggatcacca aaaacggaac
     2701 gttgagctgg acggaaatag tggtaaagtg acatgattat agtttgaaga tttctaattt
     2761 cacaattaga gcaaatgttg ttcggtattt attttcaacg gtatttatac tattttccac
     2821 ctttttctag aacattcgag ctgcttgttg caaaaggagg gcgactcaca ttcggtacat
     2881 ggaaaagtag tgtacacaat aaagagaccc agatacattt tccgtctgcg tctctttgca
     2941 cccaccggga gtattttcaa acgaatgcat ctaggacctt ctagaacatt ctgtaaggct
     3001 gcagaatgcg ggtatataag gaaagcgggc tcagaggaag ccaacacgct ttgttctagt
     3061 gcatctaaaa aacttcgaaa atcctcatcg gatcctatcg attcgcggcc gctgtacacc
     3121 cgggtgccct atagtgagtc gtattggtac cggtagaaaa aatatgcgct ttgatttcca
     3181 aaacttaaaa ttttcgatga ttttcatatt tttgtggaac atcatagtcg gatttttact
     3241 tgctcttgta aagattttca aaatctacat gtatcttgat ctgcctgaaa atgtatgggt
     3301 aaatcggttt gcgcatgtta ttgctcttat tggatcaatt gcttcattgt ggttaatttt
     3361 ttttagccca tttgaaatta agcaaggaag attttctgtc tcgtcaaaaa ctatcctgct
     3421 tcttgtttgc aagggtttga ttggcttgat atttcttgca cagattttcg cccatgtcct
     3481 ttacattgac ccatgccaga atatgctctt atcattgaac tctgcactgt tctccttgct
     3541 gctgaattat tatgagtctc gtggtgcgat ccatccgttg atggaattag ttcaaacagc
     3601 tggcctgaga ccaactttgt gcgctgctgt acgatttaat tgtctcaact caactgctca
     3661 tattgatcca agtgttgaaa atccatgaga taaatgcaaa atcctttcaa gcattccctt
     3721 cttctctatc actcttcttt ctttttgtca aaaaattctc tcgctaattt atttgctttt
     3781 ttaatgttat tattttatga ctttttatag tcactgaaaa gtttgcatct gagtgaagtg
     3841 aatgctatca aaatgtgatt ctgtctgatg tactttcaca atctctcttc aattccattt
     3901 tgaagtgctt taaacccgaa aggttgagaa aaatgcgagc gctcaaatat ttgtattgtg
     3961 ttcgttgagt gacccaacaa aaagaggaaa ctttattgtg ccgccaagaa aaaagtctca
     4021 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     4081 aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4141 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     4201 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4261 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4321 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4381 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4441 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4501 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4561 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4621 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4681 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4741 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4801 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4861 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4921 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4981 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5041 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5101 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5161 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5221 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5281 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5341 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5401 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5461 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5521 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5581 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5641 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5701 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5761 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5821 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5881 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5941 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6001 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6061 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6121 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6181 ctggtctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6241 gataaccgta ttaccgcctt tgagtgagct gatacc
//
