LOCUS       sequence-60686-e        6703 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 6703)
  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 6703)
  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: pCFJ906
FEATURES             Location/Qualifiers
     source          1..6703
                     /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"
     primer_bind     4707..4729
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4767..4785)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4853..4957
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4958..5026,5027..5818)
                     /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(5176..5195)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5989..6577
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6478..6497
                     /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 cacatccccc
     4441 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     4501 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4561 ttcacaccgc atatggtgca ctctcagtgc aatctgctct gatgccgcat agttaagcca
     4621 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4681 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4741 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4801 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4861 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4921 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4981 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5041 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5101 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5161 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5221 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5281 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5341 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5401 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5461 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5521 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5581 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5641 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5701 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5761 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5821 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5881 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5941 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6001 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6061 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6121 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6181 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6241 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6301 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6361 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6421 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6481 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6541 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6601 acggttcctg gtcttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6661 ttctgtggat aaccgtatta ccgcctttga gtgagctgat acc
//
