LOCUS       sequence-60695-e        8389 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 8389)
  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 8389)
  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: pCFJ1002
FEATURES             Location/Qualifiers
     source          1..8389
                     /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             2862..3656
                     /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(2916..2935)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3526..3545
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4357..4373
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4358..4374
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5807..6130
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(6274..6293)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6393..6415
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6453..6471)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6539..6643
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6644..6712,6713..7504)
                     /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(6862..6881)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7675..8263
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8164..8183
                     /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 tttacaggcc ggatgggcca tggcaccatt tctaaatttt agtttaataa aaaaatttta
     2101 aaataatacg agttttgaga ctaactttca ataaaaacca aaaaattgct tcaaaaaaga
     2161 gctcgcgccg atttacgata aacacagcgc gctacgacac tccgaaaacc gcgccgcacc
     2221 attacattct ccgccccacc gcacggtttg cggaaggcga tgaaattgct gggcctaaaa
     2281 ttttaaccct ttttttcccg tttttcaact gttttcccga ttttttcgat atttttttca
     2341 ggtttttagt tagaaaagta tataaaaatt gcattatttg tagttttcgc tcgtaatttg
     2401 tgatagaaaa cggccaattt gcctagattt gtgattctac caagtggaat ttttgggaaa
     2461 atatcaatat tccacgattt ttattcaaat cataacgttt cgaaattatc agcttaaata
     2521 aaactaaaat aaataataaa atactaaaaa ccctgaattt ctcggttctc ggttcagctt
     2581 gagttcgttc tatacacgtt atcgattttt gtcatttcgt ttaatcatgg gttttgattc
     2641 attttaaata tgaaatttga tttattcctc gtttcgaagt tggtttgtct tgtccggccg
     2701 gtaaacaaat gttattaaaa aaaataataa atttcgaaat ttttcgtaat gttccgcaat
     2761 cgtgtttttc cgtttatctc gtcttccctt ccttcaaata tactatttat tttctcaatt
     2821 cacatatttt cagccagccc tgctcaacaa gtggaataaa aatgattgaa caagatggat
     2881 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     2941 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3001 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     3061 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3121 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3181 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3241 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3301 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcac
     3361 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     3421 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3481 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3541 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     3601 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagtcc
     3661 aattactctt caacatccct acatgctctt tctccctgtg ctcccacccc ctatttttgt
     3721 tattatcaaa aaacttctct taatttcttt gttttttagc ttcttttaag ccacctctaa
     3781 caatgaaatt gtgtagattc aaaaatagaa ttaattcgta ataaaaagtc gaaaaaaatt
     3841 gtgctccctc cccccattaa taataattct atcccaaaat ctacacaatg ttctgtgtac
     3901 acttcttatg ttttttactt ctgataaatt tttttgaaac atcatagaaa aaaccgcaca
     3961 caaaatacct tatcatatgt tacgtttcag tttatgaccg caatttttat ttcttcgcac
     4021 gtctgggcct ctcatgacgt caaatcatgc tcatcgtgaa aaagttttgg agtatttttg
     4081 gaatttttca atcaagtgaa agtttatgaa attaattttc ctgcttttgc tttttggggt
     4141 ttcccctatt gtttgtcaag atttcgagga cggcgttttt cttgctaaaa tcacaagtat
     4201 tgatgagcac gatgcaagaa agatcggaag aaggtttggg tttgaggctc agtggaaggt
     4261 gagtagaagt tgataatttg aaagtggagt agtgtctatg gggtttttgc cttaaatgac
     4321 agaatacatt cccaatatac caaacataac tgtttctcga ggtcgacggt atcgataagc
     4381 ttgatccgga tctgcggctt actcaccaga cctggcccca tccgattacc acctattcgc
     4441 ttcgatggga cacgcactcg ctgagcagcg cttcgattct tacgaaagtg tgaaaaaatg
     4501 gctcgatgaa tggttcgccg caaaagacga tgagttctac tggcgtggaa tccacaaatt
     4561 gcccgagaga tgggaaaaat gtgtagctag cgacggcaaa tactttgaat aaatgatttt
     4621 ttctttttcc acaaaattta acgtgttttt tgatttaaaa aaaacgacat ttcatacttg
     4681 tacacctgat atctagatac ctaggtgagc tctggtaccc ccctggcgtt acccaactta
     4741 atcgccttgc agcagagtcg tattacgtag cccggcctgc aggtcgactc tagaggatca
     4801 ccaaaaacgg aacgttgagc tggacggaaa tagtggtaaa gtgacatgat tatagtttga
     4861 agatttctaa tttcacaatt agagcaaatg ttgttcggta tttattttca acggtattta
     4921 tactattttc cacctttttc tagaacattc gagctgcttg ttgcaaaagg agggcgactc
     4981 acattcggta catggaaaag tagtgtacac aataaagaga cccagataca ttttccgtct
     5041 gcgtctcttt gcacccaccg ggagtatttt caaacgaatg catctaggac cttctagaac
     5101 attctgtaag gctgcagaat gcgggtatat aaggaaagcg ggctcagagg aagccaacac
     5161 gctttgttct agtgcatcta aaaaacttcg aaaatcctca tcggatccta tcgattcgcg
     5221 gccgctgtac acccgggtgc cctatagtga gtcgtattgg taccggtaga aaaaatatgc
     5281 gctttgattt ccaaaactta aaattttcga tgattttcat atttttgtgg aacatcatag
     5341 tcggattttt acttgctctt gtaaagattt tcaaaatcta catgtatctt gatctgcctg
     5401 aaaatgtatg ggtaaatcgg tttgcgcatg ttattgctct tattggatca attgcttcat
     5461 tgtggttaat tttttttagc ccatttgaaa ttaagcaagg aagattttct gtctcgtcaa
     5521 aaactatcct gcttcttgtt tgcaagggtt tgattggctt gatatttctt gcacagattt
     5581 tcgcccatgt cctttacatt gacccatgcc agaatatgct cttatcattg aactctgcac
     5641 tgttctcctt gctgctgaat tattatgagt ctcgtggtgc gatccatccg ttgatggaat
     5701 tagttcaaac agctggcctg agaccaactt tgtgcgctgc tgtacgattt aattgtctca
     5761 actcaactgc tcatattgat ccaagtgttg aaaatccatg agataaatgc aaaatccttt
     5821 caagcattcc cttcttctct atcactcttc tttctttttg tcaaaaaatt ctctcgctaa
     5881 tttatttgct tttttaatgt tattatttta tgacttttta tagtcactga aaagtttgca
     5941 tctgagtgaa gtgaatgcta tcaaaatgtg attctgtctg atgtactttc acaatctctc
     6001 ttcaattcca ttttgaagtg ctttaaaccc gaaaggttga gaaaaatgcg agcgctcaaa
     6061 tatttgtatt gtgttcgttg agtgacccaa caaaaagagg aaactttatt gtgccgccaa
     6121 gaaaaaagtc tcaagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     6181 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg
     6241 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     6301 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     6361 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     6421 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     6481 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     6541 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     6601 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     6661 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6721 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     6781 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     6841 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     6901 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     6961 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     7021 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     7081 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     7141 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     7201 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     7261 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     7321 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     7381 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     7441 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     7501 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     7561 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     7621 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     7681 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     7741 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     7801 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     7861 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     7921 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7981 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     8041 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     8101 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     8161 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     8221 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     8281 ctttttacgg ttcctggtct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     8341 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgatacc
//
