LOCUS       sequence-60720-e        6839 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 6839)
  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 6839)
  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: pCFJ1202
FEATURES             Location/Qualifiers
     source          1..6839
                     /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"
     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"
     CDS             1312..2106
                     /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(1366..1385)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1976..1995
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2808..2824
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      4257..4580
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(4724..4743)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4843..4865
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4903..4921)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4989..5093
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5094..5162,5163..5954)
                     /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(5312..5331)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6125..6713
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6614..6633
                     /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 aggccggatg ggccatggca ccttaacatt tctaaatttt agtttaataa
      541 aaaaatttta aaataatacg agttttgaga ctaactttca ataaaaacca aaaaattgct
      601 tcaaaaaaga gctcgcgccg atttacgata aacacagcgc gctacgacac tccgaaaacc
      661 gcgccgcacc attacattct ccgccccacc gcacggtttg cggaaggcga tgaaattgct
      721 gggcctaaaa ttttaaccct ttttttcccg tttttcaact gttttcccga ttttttcgat
      781 atttttttca ggtttttagt tagaaaagta tataaaaatt gcattatttg tagttttcgc
      841 tcgtaatttg tgatagaaaa cggccaattt gcctagattt gtgattctac caagtggaat
      901 ttttgggaaa atatcaatat tccacgattt ttattcaaat cataacgttt cgaaattatc
      961 agcttaaata aaactaaaat aaataataaa atactaaaaa ccctgaattt ctcggttctc
     1021 ggttcagctt gagttcgttc tatacacgtt atcgattttt gtcatttcgt ttaatcatgg
     1081 gttttgattc attttaaata tgaaatttga tttattcctc gtttcgaagt tggtttgtct
     1141 tgtccggccg gtaaacaaat gttattaaaa aaaataataa atttcgaaat ttttcgtaat
     1201 gttccgcaat cgtgtttttc cgtttatctc gtcttccctt ccttcaaata tactatttat
     1261 tttctcaatt cacatatttt cagccagccc tgctcaacaa gtggaataaa aatgattgaa
     1321 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     1381 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     1441 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     1501 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     1561 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     1621 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     1681 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     1741 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     1801 gggctcgcac cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     1861 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     1921 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     1981 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     2041 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     2101 ttctgagtcc aattactctt caacatccct acatgctctt tctccctgtg ctcccacccc
     2161 ctatttttgt tattatcaaa aaacttctct taatttcttt gttttttagc ttcttttaag
     2221 ccacctctaa caatgaaatt gtgtagattc aaaaatagaa ttaattcgta ataaaaagtc
     2281 gaaaaaaatt gtgctccctc cccccattaa taataattct atcccaaaat ctacacaatg
     2341 ttctgtgtac acttcttatg ttttttactt ctgataaatt tttttgaaac atcatagaaa
     2401 aaaccgcaca caaaatacct tatcatatgt tacgtttcag tttatgaccg caatttttat
     2461 ttcttcgcac gtctgggcct ctcatgacgt caaatcatgc tcatcgtgaa aaagttttgg
     2521 agtatttttg gaatttttca atcaagtgaa agtttatgaa attaattttc ctgcttttgc
     2581 tttttggggt ttcccctatt gtttgtcaag atttcgagga cggcgttttt cttgctaaaa
     2641 tcacaagtat tgatgagcac gatgcaagaa agatcggaag aaggtttggg tttgaggctc
     2701 agtggaaggt gagtagaagt tgataatttg aaagtggagt agtgtctatg gggtttttgc
     2761 cttaaatgac agaatacatt cccaatatac caaacataac tgtttctcga ggtcgacggt
     2821 atcgataagc ttgatccgga tctgcggctt actcaccaga cctggcccca tccgattacc
     2881 acctattcgc ttcgatggga cacgcactcg ctgagcagcg cttcgattct tacgaaagtg
     2941 tgaaaaaatg gctcgatgaa tggttcgccg caaaagacga tgagttctac tggcgtggaa
     3001 tccacaaatt gcccgagaga tgggaaaaat gtgtagctag cgacggcaaa tactttgaat
     3061 aaatgatttt ttctttttcc acaaaattta acgtgttttt tgatttaaaa aaaacgacat
     3121 ttcatacttg tacacctgat atctagatac ctaggtgagc tctggtaccc ccctggcgtt
     3181 acccaactta atcgccttgc agcagagtcg tattacgtag cccggcctgc aggtcgactc
     3241 tagaggatca ccaaaaacgg aacgttgagc tggacggaaa tagtggtaaa gtgacatgat
     3301 tatagtttga agatttctaa tttcacaatt agagcaaatg ttgttcggta tttattttca
     3361 acggtattta tactattttc cacctttttc tagaacattc gagctgcttg ttgcaaaagg
     3421 agggcgactc acattcggta catggaaaag tagtgtacac aataaagaga cccagataca
     3481 ttttccgtct gcgtctcttt gcacccaccg ggagtatttt caaacgaatg catctaggac
     3541 cttctagaac attctgtaag gctgcagaat gcgggtatat aaggaaagcg ggctcagagg
     3601 aagccaacac gctttgttct agtgcatcta aaaaacttcg aaaatcctca tcggatccta
     3661 tcgattcgcg gccgctgtac acccgggtgc cctatagtga gtcgtattgg taccggtaga
     3721 aaaaatatgc gctttgattt ccaaaactta aaattttcga tgattttcat atttttgtgg
     3781 aacatcatag tcggattttt acttgctctt gtaaagattt tcaaaatcta catgtatctt
     3841 gatctgcctg aaaatgtatg ggtaaatcgg tttgcgcatg ttattgctct tattggatca
     3901 attgcttcat tgtggttaat tttttttagc ccatttgaaa ttaagcaagg aagattttct
     3961 gtctcgtcaa aaactatcct gcttcttgtt tgcaagggtt tgattggctt gatatttctt
     4021 gcacagattt tcgcccatgt cctttacatt gacccatgcc agaatatgct cttatcattg
     4081 aactctgcac tgttctcctt gctgctgaat tattatgagt ctcgtggtgc gatccatccg
     4141 ttgatggaat tagttcaaac agctggcctg agaccaactt tgtgcgctgc tgtacgattt
     4201 aattgtctca actcaactgc tcatattgat ccaagtgttg aaaatccatg agataaatgc
     4261 aaaatccttt caagcattcc cttcttctct atcactcttc tttctttttg tcaaaaaatt
     4321 ctctcgctaa tttatttgct tttttaatgt tattatttta tgacttttta tagtcactga
     4381 aaagtttgca tctgagtgaa gtgaatgcta tcaaaatgtg attctgtctg atgtactttc
     4441 acaatctctc ttcaattcca ttttgaagtg ctttaaaccc gaaaggttga gaaaaatgcg
     4501 agcgctcaaa tatttgtatt gtgttcgttg agtgacccaa caaaaagagg aaactttatt
     4561 gtgccgccaa gaaaaaagtc tcaagctggc gtaatagcga agaggcccgc accgatcgcc
     4621 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta
     4681 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     4741 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4801 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4861 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     4921 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     4981 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5041 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5101 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5161 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5221 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5281 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5341 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5401 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5461 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5521 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5581 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5641 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5701 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5761 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5821 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5881 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5941 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6001 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6061 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6121 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6181 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6241 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     6301 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6361 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6421 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6481 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6541 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6601 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6661 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6721 gcaacgcggc ctttttacgg ttcctggtct tttgctggcc ttttgctcac atgttctttc
     6781 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgatacc
//
