LOCUS       sequence-60693-e        8067 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 8067)
  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 8067)
  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: pCFJ1001
FEATURES             Location/Qualifiers
     source          1..8067
                     /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..442
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(483..788)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(585..604)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1130..1789)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1704..1723
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(1843..1849,1850..1867,1868..1873))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1898..2021)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2029..2046)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2029..2045)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2051..2062)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      5485..5808
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(5952..5971)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6071..6093
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6131..6149)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6217..6321
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6322..6390,6391..7182)
                     /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(6540..6559)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7353..7941
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7842..7861
                     /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 acgagaaacg taaaatgata
      361 taaatatcaa tatattaaat tagattttgc ataaaaaaca gactacataa tactgtaaaa
      421 cacaacatat ccagtcacta tggtcgacct gcagactggc tgtgtataag ggagcctgac
      481 atttatattc cccagaacat caggttaatg gcgtttttga tgtcattttc gcggtggctg
      541 agatcagcca cttcttcccc gataacggag accggcacac tggccatatc ggtggtcatc
      601 atgcgccagc tttcatcccc gatatgcacc accgggtaaa gttcacgggg gactttatct
      661 gacagcagac gtgcactggc cagggggatc accatccgtc gcccgggcgt gtcaataata
      721 tcactctgta catccacaaa cagacgataa cggctctctc ttttataggt gtaaacctta
      781 aactgcattt caccagcccc tgttctcgtc ggcaaaagag ccgttcattt caataaaccg
      841 ggcgacctca gccatccctt cctgattttc cgctttccag cgttcggcac gcagacgacg
      901 ggcttcattc tgcatggttg tgcttaccga accggagata ttgacatcat atatgccttg
      961 agcaactgat agctgtcgct gtcaactgtc actgtaatac gctgcttcat agcatacctc
     1021 tttttgacat acttcgggta tacatatcag tatatattct tataccgcaa aaatcagcgc
     1081 gcaaatacgc atactgttat ctggctttta gtaagccgga tcctctagat tacgccccgc
     1141 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
     1201 cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
     1261 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
     1321 caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc
     1381 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta
     1441 gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct
     1501 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
     1561 ttgccatacg gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg
     1621 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa
     1681 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat
     1741 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
     1801 tagctcctga aaatctcgac ggatcctaac tcaaaatcca cacattatac gagccggaag
     1861 cataaagtgt aaagcctggg gtgcctaatg cggccgccat agtgactgga tatgttgtgt
     1921 tttacagtat tatgtagtct gttttttatg caaaatctaa tttaatatat tgatatttat
     1981 atcattttac gtttctcgtt caactttatt atacatagtt gataattcac tggccgtcgt
     2041 tttacatggc acgcccttct attacagggt ttctgataac aaaccggtta ttaactccca
     2101 acaagggatg atttcaattc atcatgctca aattgaccca aattaagtta catgacaaat
     2161 tcatcgccct tttcaactct tcttggctca tcatctgtta ttctgttcct attctgtctg
     2221 caccctatac ccctttgcat actctctcgt cattcctttc ttggatagtg tcttcttctc
     2281 cccagctctg ctacttctat gacttgcgcc cgctgctttt ccgctcgttc ctctcctctc
     2341 tgacgtcttc ttctgcttct tttctccttc ctttttccct tctcaatctt cctctttttc
     2401 ccatttctgt caatcattcg aagaagaaga agaagaagac cctcattcat ttcatttttt
     2461 tttctgtcgg tgtgtgctgc tcggttaaga gtgagctctc tattccacgt cttcttcttt
     2521 ttcttcttcg attcgaatca atcactccac caaaacgcat tcgtttttgg gattcacccc
     2581 gcggttcgca ataggtttct ttttcaaatt attagcgtta taaatagaaa aatggtggag
     2641 tttcaataaa aatgataatt tacaaagtgt attttgatta catgtactca aaaaggttga
     2701 aatttttcat accagttttc cggaaatcca tctgatatca ttatcgtatt tttcttttta
     2761 aaaatgtttt caaaaaaaaa caaaatatag ctggttattt ggcaccctct aattaccatt
     2821 ttcctgtcac accacactct tttccttctt ccacttcttt ccgttttcag ccgcttccaa
     2881 ccaaaccgat atgaaagccg agcaacaacg atcgattcca cccggctcgg cgaccttccc
     2941 gtcgcaggtg agactcagaa aactagagaa accgctcaac taactcttga tatccgattt
     3001 cattcttttc tcttttcttt ttgttgaact tttccatatt tccagatgcc acggccacca
     3061 ccaagcaccg aacaaggaat cacaacggaa tcggagcttg cgaagaaagc tcaaatcact
     3121 ccgaacgacg ttttagcact tccgggaatc actcaaggta tcttgctccc tttggatttc
     3181 agtttaaaac ataaatttag gattcttatg ctccccatct gcgaacatct ataacatcga
     3241 gttcaccaag ttccaaatcc gtgatctgga cactgagcaa gtgcttttcg agatcgccaa
     3301 accggagaac gatcaggaga atgatgagtc gccacaggag tcggcaagat acgtgcgtta
     3361 tagatttgct ccaaactttt tgaaactcaa aacggtcgga gcaactgtgg aattcaaagt
     3421 aggagacatc ccaatccatc atttccgaat gatcgaacgt cacttcttca aagatcgcct
     3481 tctgaagtgt tttgactttg aattcggatt ctgtattccg aattcacgaa acaactgtga
     3541 acatatctat gagttccctc aactctctca acaactcagt gagtcattat tctaaaaagt
     3601 acaattcaaa agactaatct ctttttcagt ggacgacatg atcaacaatc caaacgagac
     3661 tcgttctgac agcttctatt tcgtcgataa caaactcgtc atgcacaaca aagccgacta
     3721 ctcatatgat gcataaatat ttaatacaaa aatgttctgg ataattattc tgtcgaatag
     3781 aaaaaaaact ccaaatgtga ttaaattcca ataattcctg tctagtttgt tccttcccct
     3841 tccccttctc atgttcaatg cattcctaag cttttcagtt ccccccttgt ttctatattt
     3901 tttcgctgtc ctgtcacact cgctaaaaac actaatcaca cggaaatctg ttttcaataa
     3961 aaactccaac tttaactcat tttcaatttc aactgaaaga ttttttcatt agagaatgtc
     4021 tagaactagg cccgggctac gtaatacgac tcacttaagg gtgccggatc tgcggcttac
     4081 tcaccagacc tggccccatc cgattaccac ctattcgctt cgatgggaca cgcactcgct
     4141 gagcagcgct tcgattctta cgaaagtgtg aaaaaatggc tcgatgaatg gttcgccgca
     4201 aaagacgatg agttctactg gcgtggaatc cacaaattgc ccgagagatg ggaaaaatgt
     4261 gtagctagcg acggcaaata ctttgaataa atgatttttt ctttttccac aaaatttaac
     4321 gtgttttttg atttaaaaaa aacgacattt catacttgta cacctgatat ctagatacct
     4381 aggtgagctc tggtaccccc ctggcgttac ccaacttaat cgccttgcag cagagtcgta
     4441 ttacgtagcc cggcctgcag gtcgactcta gaggatcacc aaaaacggaa cgttgagctg
     4501 gacggaaata gtggtaaagt gacatgatta tagtttgaag atttctaatt tcacaattag
     4561 agcaaatgtt gttcggtatt tattttcaac ggtatttata ctattttcca cctttttcta
     4621 gaacattcga gctgcttgtt gcaaaaggag ggcgactcac attcggtaca tggaaaagta
     4681 gtgtacacaa taaagagacc cagatacatt ttccgtctgc gtctctttgc acccaccggg
     4741 agtattttca aacgaatgca tctaggacct tctagaacat tctgtaaggc tgcagaatgc
     4801 gggtatataa ggaaagcggg ctcagaggaa gccaacacgc tttgttctag tgcatctaaa
     4861 aaacttcgaa aatcctcatc ggatcctatc gattcgcggc cgctgtacac ccgggtgccc
     4921 tatagtgagt cgtattggta ccggtagaaa aaatatgcgc tttgatttcc aaaacttaaa
     4981 attttcgatg attttcatat ttttgtggaa catcatagtc ggatttttac ttgctcttgt
     5041 aaagattttc aaaatctaca tgtatcttga tctgcctgaa aatgtatggg taaatcggtt
     5101 tgcgcatgtt attgctctta ttggatcaat tgcttcattg tggttaattt tttttagccc
     5161 atttgaaatt aagcaaggaa gattttctgt ctcgtcaaaa actatcctgc ttcttgtttg
     5221 caagggtttg attggcttga tatttcttgc acagattttc gcccatgtcc tttacattga
     5281 cccatgccag aatatgctct tatcattgaa ctctgcactg ttctccttgc tgctgaatta
     5341 ttatgagtct cgtggtgcga tccatccgtt gatggaatta gttcaaacag ctggcctgag
     5401 accaactttg tgcgctgctg tacgatttaa ttgtctcaac tcaactgctc atattgatcc
     5461 aagtgttgaa aatccatgag ataaatgcaa aatcctttca agcattccct tcttctctat
     5521 cactcttctt tctttttgtc aaaaaattct ctcgctaatt tatttgcttt tttaatgtta
     5581 ttattttatg actttttata gtcactgaaa agtttgcatc tgagtgaagt gaatgctatc
     5641 aaaatgtgat tctgtctgat gtactttcac aatctctctt caattccatt ttgaagtgct
     5701 ttaaacccga aaggttgaga aaaatgcgag cgctcaaata tttgtattgt gttcgttgag
     5761 tgacccaaca aaaagaggaa actttattgt gccgccaaga aaaaagtctc aagctggcgt
     5821 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     5881 tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     5941 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     6001 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     6061 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     6121 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     6181 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     6241 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6301 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6361 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6421 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6481 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6541 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6601 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6661 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6721 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6781 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6841 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6901 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6961 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     7021 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     7081 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     7141 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     7201 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     7261 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7321 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7381 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7441 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     7501 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7561 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7621 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7681 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7741 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7801 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7861 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7921 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggtcttt
     7981 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     8041 attaccgcct ttgagtgagc tgatacc
//
