LOCUS       sequence-60692-e        7851 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 7851)
  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 7851)
  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: pCFJ1000
FEATURES             Location/Qualifiers
     source          1..7851
                     /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(1108..1788)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     1703..1722
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(1842..1848,1849..1866,1867..1872))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1897..2020)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2028..2045)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2028..2044)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2658..2677)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3256..3276
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     terminator      5269..5592
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(5736..5755)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5855..5877
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5915..5933)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6001..6105
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6106..6174,6175..6966)
                     /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(6324..6343)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7137..7725
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7626..7645
                     /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 ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc
     1201 acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
     1261 atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
     1321 aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc
     1381 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
     1441 aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc
     1501 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
     1561 tgccatacgg aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
     1621 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
     1681 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
     1741 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
     1801 agctcctgaa aatctcgacg gatcctaact caaaatccac acattatacg agccggaagc
     1861 ataaagtgta aagcctgggg tgcctaatgc ggccgccata gtgactggat atgttgtgtt
     1921 ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata
     1981 tcattttacg tttctcgttc aactttatta tacatagttg ataattcact ggccgtcgtt
     2041 ttacagaggg gagaaaatgc ggagagaaga acccagcatc ttaataaaat tatataattt
     2101 tgaataaaat caaattaaaa ataagattaa ttgaaacaca acaatagaat gcagaaaatt
     2161 actcataaga aataatttat tttgaaagaa gttgtgtcga tttacgataa aaactacacg
     2221 caaacgaggc tccgcgttta caccgtttgg caatggagcg cgcttgctat tagattttag
     2281 gccagctcaa cgaaagttta atttattttt cttctttttt tccatttttt tgagtaaaat
     2341 atcgaatttt ttgtattaat tctgtaaatt ttataatttg caactatttt ggcgtattct
     2401 gcttgttttt atcagcgaat ttttatgcat ttcagtcaaa agtttgagca ggaaaaatga
     2461 catttaggac tttgtaaaat tgtattctta aatttgcttg aaaacctacg tgtacaattg
     2521 attttattga atttttctta cgtttcccgt tgtatcttat tgtatttaac tttcatcata
     2581 aaataattta tttccaggta aaatatccga cgctctcgtg gatccccggg attggctcag
     2641 aaaaaaaggt accgaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     2701 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     2761 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     2821 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcggtaa gtttaaacat atatatacta
     2881 actaaccctg attatttaaa ttttcagcgg acgacggcgc cgcggtggcg gtctggacca
     2941 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
     3001 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
     3061 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgacgta agtttaaaca
     3121 tgattttact aactaactaa tctgatttaa attttcagca ccagggcaag ggtctgggca
     3181 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     3241 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     3301 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgat
     3361 gagctagccg gccatacaag taatccggat gatcgacgcc aacgtcgttg aattttcaaa
     3421 ttttaaatac tgaatatttg ttttttttcc tattatttat ttattctctt tgtgtttttt
     3481 ttcttgcttt ctaaaaaatt aattcaatcc aaatctaaac attttttttt ctctttccgt
     3541 ctcccaattc gtattccgct cctctcatct gaacacaatg tgcaagttta tttatcttct
     3601 cgctttcatt tcattaggac gtggggggaa ttggtggaag ggggaaacac acaaaaggat
     3661 gatggaaatg aaataaggac acacaatatg caacaacatt caattcagaa atatggagga
     3721 aggtttaaaa gaaaacataa aaatatatag aggaggaagg aaaggtgccg gatctgcggc
     3781 ttactcacca gacctggccc catccgatta ccacctattc gcttcgatgg gacacgcact
     3841 cgctgagcag cgcttcgatt cttacgaaag tgtgaaaaaa tggctcgatg aatggttcgc
     3901 cgcaaaagac gatgagttct actggcgtgg aatccacaaa ttgcccgaga gatgggaaaa
     3961 atgtgtagct agcgacggca aatactttga ataaatgatt ttttcttttt ccacaaaatt
     4021 taacgtgttt tttgatttaa aaaaaacgac atttcatact tgtacacctg atatctagat
     4081 acctaggtga gctctggtac ccccctggcg ttacccaact taatcgcctt gcagccaact
     4141 taatcaccat gcagccaact taatcgcctt gcagccaact taatcgcctt gcagccaact
     4201 taatcgcctt gcagcagagt cgtattacgt agcccggcct gcaggtcgac tctagaggat
     4261 caccaaaaac ggaacgttga gctggacgga aatagtggta aagtgacatg attatagttt
     4321 gaagatttct aatttcacaa ttagagcaaa tgttgttcgg tatttatttt caacggtatt
     4381 tatactattt tccacctttt tctagaacat tcgagctgct tgttgcaaaa ggagggcgac
     4441 tcacattcgg tacatggaaa agtagtgtac acaataaaga gacccagata cattttccgt
     4501 ctgcgtctct ttgcacccac cgggagtatt ttcaaacgaa tgcatctagg accttctaga
     4561 acattctgta aggctgcaga atgcgggtat ataaggaaag cgggctcaga ggaagccaac
     4621 acgctttgtt ctagtgcatc taaaaaactt cgaaaatcct catcggatcc tatcgattcg
     4681 cggccgctgt acacccgggt gccctatagt gagtcgtatt ggtaccggta gaaaaaatat
     4741 gcgctttgat ttccaaaact taaaattttc gatgattttc atatttttgt ggaacatcat
     4801 agtcggattt ttacttgctc ttgtaaagat tttcaaaatc tacatgtatc ttgatctgcc
     4861 tgaaaatgta tgggtaaatc ggtttgcgca tgttattgct cttattggat caattgcttc
     4921 attgtggtta atttttttta gcccatttga aattaagcaa ggaagatttt ctgtctcgtc
     4981 aaaaactatc ctgcttcttg tttgcaaggg tttgattggc ttgatatttc ttgcacagat
     5041 tttcgcccat gtcctttaca ttgacccatg ccagaatatg ctcttatcat tgaactctgc
     5101 actgttctcc ttgctgctga attattatga gtctcgtggt gcgatccatc cgttgatgga
     5161 attagttcaa acagctggcc tgagaccaac tttgtgcgct gctgtacgat ttaattgtct
     5221 caactcaact gctcatattg atccaagtgt tgaaaatcca tgagataaat gcaaaatcct
     5281 ttcaagcatt cccttcttct ctatcactct tctttctttt tgtcaaaaaa ttctctcgct
     5341 aatttatttg cttttttaat gttattattt tatgactttt tatagtcact gaaaagtttg
     5401 catctgagtg aagtgaatgc tatcaaaatg tgattctgtc tgatgtactt tcacaatctc
     5461 tcttcaattc cattttgaag tgctttaaac ccgaaaggtt gagaaaaatg cgagcgctca
     5521 aatatttgta ttgtgttcgt tgagtgaccc aacaaaaaga ggaaacttta ttgtgccgcc
     5581 aagaaaaaag tctcaagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     5641 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
     5701 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag
     5761 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     5821 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     5881 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     5941 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     6001 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     6061 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     6121 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6181 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     6241 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     6301 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     6361 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     6421 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6481 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     6541 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6601 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6661 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6721 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6781 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6841 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6901 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6961 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     7021 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     7081 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     7141 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7201 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7261 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     7321 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7381 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7441 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7501 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7561 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7621 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7681 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7741 gcctttttac ggttcctggt cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     7801 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac c
//
