LOCUS       sequence-60709-e        4728 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 4728)
  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 4728)
  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: pCFJ1258
FEATURES             Location/Qualifiers
     source          1..4728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             362..373
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    403..526
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(551..556,557..574,575..581)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             635..1294
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(701..720)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1636..1941
                     /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     1820..1839
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1982..2105)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2125..2141)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2125..2141)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(2550..3138)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2630..2649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3309..4100,4101..4169))
                     /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     3932..3951
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4170..4274)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4342..4360
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4398..4420)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4520..4539
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gggtaccaga gctcacctag gtatctagat atcaggtgta caagtatgaa atgtcgtttt
       61 ttttaaatca aaaaacacgt taaattttgt ggaaaaagaa aaaatcattt attcaaagta
      121 tttgccgtcg ctagctacac atttttccca tctctcgggc aatttgtgga ttccacgcca
      181 gtagaactca tcgtcttttg cggcgaacca ttcatcgagc cattttttca cactttcgta
      241 agaatcgaag cgctgctcag cgagtgcgtg tcccatcgaa gcgaataggt ggtaatcgga
      301 tggggccagg tctggtgagt aagccgcaga tccggcaccc tttgttatca gaaaccctgt
      361 aatagaaggg cgtgccatgt aaaacgacgg ccagtgaatt atcaactatg tataataaag
      421 ttgaacgaga aacgtaaaat gatataaata tcaatatatt aaattagatt ttgcataaaa
      481 aacagactac ataatactgt aaaacacaac atatccagtc actatggcgg ccgcattagg
      541 caccccaggc tttacacttt atgcttccgg ctcgtataat gtgtggattt tgagttagga
      601 tccgtcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac
      661 caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc
      721 tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa
      781 gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc
      841 tcatccggaa ttccgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca
      901 cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata
      961 ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga
     1021 aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc
     1081 ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc
     1141 cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat
     1201 tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca
     1261 acagtactgc gatgagtggc agggcggggc gtaatctaga ggatccggct tactaaaagc
     1321 cagataacag tatgcgtatt tgcgcgctga tttttgcggt ataagaatat atactgatat
     1381 gtatacccga agtatgtcaa aaagaggtat gctatgaagc agcgtattac agtgacagtt
     1441 gacagcgaca gctatcagtt gctcaaggca tatatgatgt caatatctcc ggttcggtaa
     1501 gcacaaccat gcagaatgaa gcccgtcgtc tgcgtgccga acgctggaaa gcggaaaatc
     1561 aggaagggat ggctgaggtc gcccggttta ttgaaatgaa cggctctttt gccgacgaga
     1621 acaggggctg gtgaaatgca gtttaaggtt tacacctata aaagagagag ccgttatcgt
     1681 ctgtttgtgg atgtacagag tgatattatt gacacgcccg ggcgacggat ggtgatcccc
     1741 ctggccagtg cacgtctgct gtcagataaa gtcccccgtg aactttaccc ggtggtgcat
     1801 atcggggatg aaagctggcg catgatgacc accgatatgg ccagtgtgcc ggtctccgtt
     1861 atcggggaag aagtggctga tctcagccac cgcgaaaatg acatcaaaaa cgccattaac
     1921 ctgatgttct ggggaatata aatgtcaggc tcccttatac acagccagtc tgcaggtcga
     1981 ccatagtgac tggatatgtt gtgttttaca gtattatgta gtctgttttt tatgcaaaat
     2041 ctaatttaat atattgatat ttatatcatt ttacgtttct cgttcaactt ttctatacaa
     2101 agttgatagc ttggcgtaat catggtcata gctgtttcct ggatccggcc gcagctgttt
     2161 tcttcaaatg aaaacagaaa attaatactg tccgcgtttg ctctttattc ggcacgaaac
     2221 tcgacatgtt gactgcactg agagtaaaca attatgacgc tcaattcgcg ccaaactatg
     2281 gtggttcgac agtcaaggtt gacacttcac aaggtcaaag ttttatgaca atcgataaat
     2341 atttacgttt gcgagacatc tatatgttcg aaacgacatt tcatacttgt acacctgata
     2401 gaattcctcg agacgtacgg tgcggtatca gctcactcaa aggcggtaat acggttatcc
     2461 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaagaccagg
     2521 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2581 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2641 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2701 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2761 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2821 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2881 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2941 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     3001 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3061 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3121 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3181 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3241 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3301 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3361 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3421 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3481 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     3541 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3601 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3661 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3721 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3781 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3841 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3901 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3961 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4021 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4081 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4141 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4201 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4261 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     4321 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     4381 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     4441 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     4501 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     4561 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     4621 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     4681 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccaggg
//
