LOCUS       sequence-78567-e        8747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  miniMos transposable element with HygroR and peel-1, R4-R3 Gateway 
            cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8747)
  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 8747)
  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: pCFJ1656
FEATURES             Location/Qualifiers
     source          1..8747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    37..160
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(201..506)
                     /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(303..322)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(848..1507)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1422..1441
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(1561..1567,1568..1585,1586..1591))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1616..1739)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1747..1764)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1747..1763)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5883..6206
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(6350..6369)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6469..6491
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6529..6547)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6615..6719
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6720..6788,6789..7580)
                     /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(6938..6957)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7751..8339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8240..8259
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca agctatcaac tttgtataga aaagttgaac
       61 gagaaacgta aaatgatata aatatcaata tattaaatta gattttgcat aaaaaacaga
      121 ctacataata ctgtaaaaca caacatatcc agtcactatg gtcgacctgc agactggctg
      181 tgtataaggg agcctgacat ttatattccc cagaacatca ggttaatggc gtttttgatg
      241 tcattttcgc ggtggctgag atcagccact tcttccccga taacggagac cggcacactg
      301 gccatatcgg tggtcatcat gcgccagctt tcatccccga tatgcaccac cgggtaaagt
      361 tcacggggga ctttatctga cagcagacgt gcactggcca gggggatcac catccgtcgc
      421 ccgggcgtgt caataatatc actctgtaca tccacaaaca gacgataacg gctctctctt
      481 ttataggtgt aaaccttaaa ctgcatttca ccagcccctg ttctcgtcgg caaaagagcc
      541 gttcatttca ataaaccggg cgacctcagc catcccttcc tgattttccg ctttccagcg
      601 ttcggcacgc agacgacggg cttcattctg catggttgtg cttaccgaac cggagatatt
      661 gacatcatat atgccttgag caactgatag ctgtcgctgt caactgtcac tgtaatacgc
      721 tgcttcatag catacctctt tttgacatac ttcgggtata catatcagta tatattctta
      781 taccgcaaaa atcagcgcgc aaatacgcat actgttatct ggcttttagt aagccggatc
      841 ctctagatta cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc
      901 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
      961 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca
     1021 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa
     1081 acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat
     1141 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg
     1201 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca
     1261 ccagctcacc gtctttcatt gccatacgga attccggatg agcattcatc aggcgggcaa
     1321 gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg
     1381 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
     1441 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt
     1501 tctccatttt agcttcctta gctcctgaaa atctcgacgg atcctaactc aaaatccaca
     1561 cattatacga gccggaagca taaagtgtaa agcctggggt gcctaatgcg gccgccatag
     1621 tgactggata tgttgtgttt tacagtatta tgtagtctgt tttttatgca aaatctaatt
     1681 taatatattg atatttatat cattttacgt ttctcgttca actttattat acatagttga
     1741 taattcactg gccgtcgttt tacaggccgg atatgaagat gaacgcaagg agcctgatgc
     1801 agcgactctt gctaggagat catcagaaag cttatatttg attagtgacg tcgcatcaaa
     1861 gctgaacaca taaattcttt aatattttat atccccatat tatgcacctg agaggaatga
     1921 agaaatttgc tgatactccc ggtgaattca tgtcttgatg gttgtgcgca ttgtggtaat
     1981 atctgaaaaa tttcgaagtc agttttctga taaggcaaca actaataaat tgagaatcaa
     2041 tttaccctta aggaacgatg ttcgcttctg aatttattaa taaatttttc cgtaattcaa
     2101 ttcaaaattg tcatatttaa gaaaaaaaca tcaaaaatga caaattcaat attttaatta
     2161 atttttgctt tcgtcgtaaa tctacacacg cgtctcttcc gtgcgagagt ccaagccagc
     2221 agccaaattc gttgactgag tattcaacgt ttatacgttg tcggcaacga gaaataggaa
     2281 aatgcatcgg gaaatgttct tttttcgatt ttttccaagg ttttgacaaa ttttaccacg
     2341 aattttgcta tgttttcaat taaaaaatat gttattcaac tgtttctatg aggaaaataa
     2401 ggctttgcat gtaattttct tattcagcat aatttttaat taatttgaat tttctgtcct
     2461 aacgtttatt ttgttttctt ggttatgact gatctgaaat taatttttga attttaaggt
     2521 aatatgaaaa aacccgagtt gaccgccaca tccgtagaga agttcctcat cgagaagttc
     2581 gactccgtct ccgacctcat gcaactctcc gagggagagg agtcccgtgc cttctccttc
     2641 gacgtcggag gacgtggata cgtcctccgt gtcaactcct gcgccgacgg attctacaag
     2701 gtaagtttaa acatatatat actaactaac cctgattatt taaattttca ggaccgttac
     2761 gtctaccgtc acttcgcctc cgccgccctc ccaatcccag aggtcctcga catcggagag
     2821 ttctccgagt ccctcaccta ctgcatctcc cgtcgtgccc aaggagtcac cctccaagac
     2881 ctcccagaga ccgagctccc agccgtcctc caaccagtcg ccgaggccat ggacgccatc
     2941 gccgccgccg acctctccca aacctccgga ttcggaccat tcggaccaca aggaatcgga
     3001 caatacacca cctggcgtga cttcatctgc gccatcgccg acccacacgt ctaccactgg
     3061 caaaccgtca tggacgacac cgtctccgcc tccgtcgccc aagccctcga cgagctcatg
     3121 ctctgggccg aggactgccc agaggtccgt cacctcgtcc acgccgactt cggatccaac
     3181 aacgtcctca ccgacaacgg acgtatcacc gccgtcatcg actggtccga ggccatgttc
     3241 ggagactccc aatacgaggt cgccaacatc ttcttctggc gtccatggct cgcctgcatg
     3301 gagcaacaaa cccgttactt cgagcgtcgt cacccagagc tcgccggatc cccacgtctc
     3361 cgtgcctaca tgctccgtat cggactcgac caactctacc aatccctcgt cgacggaaac
     3421 ttcgacgacg ccgcctgggc ccaaggacgt tgcgacgcca tcgtccgttc cggagccgga
     3481 accgtcggac gtacccaaat cgcccgtcgt tccgccgccg tctggaccga cggatgcgtc
     3541 gaggtcctcg ccgactccgg aaaccgtcgt ccatccaccc gtccacgtgc caaggtaagt
     3601 ttaaacagtt cggtactaac taaccataca tatttaaatt ttcaggagta aaattccaac
     3661 tgagcgccgg tcgctaccat taccaacttg tctggtgtca aaaataatag gggccgctgt
     3721 catcagagta agtttaaact gagttctact aactaacgag taatatttaa attttcagca
     3781 tctcgcgccc gtgcctctga cttctaagtc caattactct tcaacatccc tacatgctct
     3841 ttctccctgt gctcccaccc cctatttttg ttattatcaa aaaacttctc ttaatttctt
     3901 tgttttttag cttcttttaa gtcacctcta acaatgaaat tgtgtagatt caaaaataga
     3961 attaattcgt aataaaaagt cgaaaaaaat tgtgctccct ccccccatta ataataattc
     4021 tatcccaaaa tctacacaat gttctgtgta cacttcttat gttttttact tctgataaat
     4081 ttttttgaaa catcatagaa aaaaccgcac acaaaatacc ttatcatatg ttacgtttca
     4141 gtttatgacc gcaattttta tttcttcgca cgtctgggcc tctcatgacg tcaaatcatg
     4201 ctcatcgtga aaaagttttg gagtattttt ggaatttttc aatcaagtga aagtttatga
     4261 aattaatttt cctgcttttg ctttttgggg tttcccctat tgtttgtcaa gatttcgagg
     4321 acggcgtttt tcttgctaaa atcacaagta ttgatgagca cgatgcaaga aagatcggaa
     4381 gaaggtttgg gtttgaggct cagtggaagg tgagtagaag ttgataattt gaaagtggag
     4441 tagtgtctat gggagcttga tccggatctg cggcttactc accagacctg gccccatccg
     4501 attaccacct attcgcttcg atgggacacg cactcgctga gcagcgcttc gattcttacg
     4561 aaagtgtgaa aaaatggctc gatgaatggt tcgccgcaaa agacgatgag ttctactggc
     4621 gtggaatcca caaattgccc gagagatggg aaaaatgtgt agctagcgac ggcaaatact
     4681 ttgaataaat gattttttct ttttccacaa aatttaacgt gttttttgat ttaaaaaaaa
     4741 cgacatttca tacttgtaca cctgatatct agatacctag gtgagctctg gtacccccct
     4801 ggcgttaccc aacttaatcg ccttgcagca gagtcgtatt acgtagcccg gcctgcaggt
     4861 cgactctaga ggatcaccaa aaacggaacg ttgagctgga cggaaatagt ggtaaagtga
     4921 catgattata gtttgaagat ttctaatttc acaattagag caaatgttgt tcggtattta
     4981 ttttcaacgg tatttatact attttccacc tttttctaga acattcgagc tgcttgttgc
     5041 aaaaggaggg cgactcacat tcggtacatg gaaaagtagt gtacacaata aagagaccca
     5101 gatacatttt ccgtctgcgt ctctttgcac ccaccgggag tattttcaaa cgaatgcatc
     5161 taggaccttc tagaacattc tgtaaggctg cagaatgcgg gtatataagg aaagcgggct
     5221 cagaggaagc caacacgctt tgttctagtg catctaaaaa acttcgaaaa tcctcatcgg
     5281 atcctatcga ttcgcggccg ctgtacaccc gggtgcccta tagtgagtcg tattggtacc
     5341 ggtagaaaaa atatgcgctt tgatttccaa aacttaaaat tttcgatgat tttcatattt
     5401 ttgtggaaca tcatagtcgg atttttactt gctcttgtaa agattttcaa aatctacatg
     5461 tatcttgatc tgcctgaaaa tgtatgggta aatcggtttg cgcatgttat tgctcttatt
     5521 ggatcaattg cttcattgtg gttaattttt tttagcccat ttgaaattaa gcaaggaaga
     5581 ttttctgtct cgtcaaaaac tatcctgctt cttgtttgca agggtttgat tggcttgata
     5641 tttcttgcac agattttcgc ccatgtcctt tacattgacc catgccagaa tatgctctta
     5701 tcattgaact ctgcactgtt ctccttgctg ctgaattatt atgagtctcg tggtgcgatc
     5761 catccgttga tggaattagt tcaaacagct ggcctgagac caactttgtg cgctgctgta
     5821 cgatttaatt gtctcaactc aactgctcat attgatccaa gtgttgaaaa tccatgagat
     5881 aaatgcaaaa tcctttcaag cattcccttc ttctctatca ctcttctttc tttttgtcaa
     5941 aaaattctct cgctaattta tttgcttttt taatgttatt attttatgac tttttatagt
     6001 cactgaaaag tttgcatctg agtgaagtga atgctatcaa aatgtgattc tgtctgatgt
     6061 actttcacaa tctctcttca attccatttt gaagtgcttt aaacccgaaa ggttgagaaa
     6121 aatgcgagcg ctcaaatatt tgtattgtgt tcgttgagtg acccaacaaa aagaggaaac
     6181 tttattgtgc cgccaagaaa aaagtctcaa gctggcgtaa tagcgaagag gcccgcaccg
     6241 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc
     6301 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct
     6361 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     6421 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     6481 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     6541 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     6601 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     6661 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     6721 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     6781 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     6841 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     6901 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     6961 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     7021 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     7081 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     7141 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     7201 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     7261 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     7321 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     7381 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     7441 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     7501 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     7561 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     7621 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     7681 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     7741 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     7801 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     7861 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     7921 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7981 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     8041 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     8101 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     8161 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     8221 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     8281 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     8341 acgccagcaa cgcggccttt ttacggttcc tggtcttttg ctggcctttt gctcacatgt
     8401 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     8461 ataccgcacc gtacgtctcg aggaattcta tcaggtgtac aagtatgaaa tgtcgtttcg
     8521 aacatataga tgtctcgcaa acgtaaatat ttatcgattg tcataaaact ttgaccttgt
     8581 gaagtgtcaa ccttgactgt cgaaccacca tagtttggcg cgaattgagc gtcataattg
     8641 tttactctca gtgcagtcaa catgtcgagt ttcgtgccga ataaagagca aacgcggaca
     8701 gtattaattt tctgttttca tttgaagaaa acagctgcgg ccggatc
//
