LOCUS       sequence-60728-e        7578 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 7578)
  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 7578)
  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: pCFJ1208
FEATURES             Location/Qualifiers
     source          1..7578
                     /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..339
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        340..939
                     /gene="eft-3"
                     /label=eft-3 promoter
                     /note="promoter for the C. elegans elongation factor 
                     1-alpha gene (Mitrovich and Anderson, 2000)"
     protein_bind    965..989
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             992..1012
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1013..2437
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL
                     VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ
                     ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH
                     LFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYE
                     GTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMN
                     FEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVL
                     KGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYER
                     SEGRHHLFLYGMDELYK"
     CDS             2447..2518
                     /codon_start=1
                     /product="nuclear localization signal from the C. elegans 
                     EGL-13 transcription factor (Lyssenko et al., 2007)"
                     /label=EGL-13 NLS
                     /translation="SRRRKANPTKLSENAKKLAKEVEN"
     protein_bind    complement(2520..2544)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      2549..2872
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     protein_bind    complement(2876..2896)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(2904..2921)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2904..2920)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2926..2937)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     5582..5604
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5642..5660)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5728..5832
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5833..5901,5902..6693)
                     /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(6051..6070)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6864..7452
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7353..7372
                     /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 gaaaagttgg cacctttggt cttttattgt
      361 caacttccat tggttcttcc attgtttctg ttaaattaat gaatttttca taaaataaag
      421 acattataca atataaaaat gaagaattta ttgaaaataa actgccagag agaaaaagta
      481 tgcaacactc ccgccgagag tgtttgaaat ggtgtacggt acattttcgt gctaggagtt
      541 agatgtgcag gcagcaacga gagggggaga gatttttttg ggccttgtga aattaacgtg
      601 agttttctgg tcatctgact aatcatgttg gttttttgtt ggtttatttt gtttttatct
      661 ttgtttttat ccagattagg aaatttaaat tttatgaatt tataatgagg tcaaacattc
      721 agtcccagcg tttttcctgt tctcactgtt tagtcgaatt tttattttag gctttcaaca
      781 aatgttctaa ctgtcttatt tgtgacctca ctttttatat ttttttaatt tttaaaaata
      841 ttagaagttt ctaggataat tttttcgact tttattctct ctaccgtccg cactcttctt
      901 acttttaaat taaattgttt ttttttcagt tgggaaacac tttgctcact ccgtagcagc
      961 catggcaagt ttgtacaaaa aagcaggctc gccaaagaag aagcgtaagg tcgtgagcaa
     1021 gggcgaggag gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg gctccatgaa
     1081 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     1141 cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc
     1201 ccagttcatg tacggctcca aggcgtacgt gaagcacccc gccgacatcc ccgattacaa
     1261 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     1321 tctggtgacc gtgacccagg actcctccct gcaggacggc acgctgatct acaaggtgaa
     1381 gatgcgcggc accaacttcc cccccgacgg ccccgtaatg cagaagaaga ccatgggctg
     1441 ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg agatccacca
     1501 ggccctgaag ctgaaggacg gcggccacta cctggtggag ttcaagacca tctacatggc
     1561 caagaagccc gtgcaactgc ccggctacta ctacgtggac accaagctgg acatcacctc
     1621 ccacaacgag gactacacca tcgtggaaca gtacgagcgc tccgagggcc gccaccacct
     1681 gttcctgggg catggcaccg gcagcaccgg cagcggcagc tccggcaccg cctcctccga
     1741 ggacaacaac atggccgtca tcaaagagtt catgcgcttc aaggtgcgca tggagggctc
     1801 catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     1861 ccagaccgcc aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct
     1921 gtccccccag ttcatgtacg gctccaaggc gtacgtgaag caccccgccg acatccccga
     1981 ttacaagaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     2041 cggcggtctg gtgaccgtga cccaggactc ctccctgcag gacggcacgc tgatctacaa
     2101 ggtgaagatg cgcggcacca acttcccccc cgacggcccc gtaatgcaga agaagaccat
     2161 gggctgggag gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagat
     2221 ccaccaggcc ctgaagctga aggacggcgg ccactacctg gtggagttca agaccatcta
     2281 catggccaag aagcccgtgc aactgcccgg ctactactac gtggacacca agctggacat
     2341 cacctcccac aacgaggact acaccatcgt ggaacagtac gagcgctccg agggccgcca
     2401 ccacctgttc ctgtacggca tggacgagct gtacaagtac ccagcttccc gtcgtcgtaa
     2461 ggccaaccca accaagctct ccgagaacgc caagaagctc gccaaggagg tcgagaacaa
     2521 cccagctttc ttgtacaaag tgggataaat gcaaaatcct ttcaagcatt cccttcttct
     2581 ctatcactct tctttctttt tgtcaaaaaa ttctctcgct aatttatttg cttttttaat
     2641 gttattattt tatgactttt tatagtcact gaaaagtttg catctgagtg aagtgaatgc
     2701 tatcaaaatg tgattctgtc tgatgtactt tcacaatctc tcttcaattc cattttgaag
     2761 tgctttaaac ccgaaaggtt gagaaaaatg cgagcgctca aatatttgta ttgtgttcgt
     2821 tgagtgaccc aacaaaaaga ggaaacttta ttgtgccgcc aagaaaaaag tctcacaact
     2881 ttattataca tagttgataa ttcactggcc gtcgttttac atggcacgcc cttctattac
     2941 agggtttctg ataacaaacc ggttattaac tcccaacaag ggatgatttc aattcatcat
     3001 gctcaaattg acccaaatta agttacatga caaattcatc gcccttttca actcttcttg
     3061 gctcatcatc tgttattctg ttcctattct gtctgcaccc tatacccctt tgcatactct
     3121 ctcgtcattc ctttcttgga tagtgtcttc ttctccccag ctctgctact tctatgactt
     3181 gcgcccgctg cttttccgct cgttcctctc ctctctgacg tcttcttctg cttcttttct
     3241 ccttcctttt tcccttctca atcttcctct ttttcccatt tctgtcaatc attcgaagaa
     3301 gaagaagaag aagaccctca ttcatttcat ttttttttct gtcggtgtgt gctgctcggt
     3361 taagagtgag ctctctattc cacgtcttct tctttttctt cttcgattcg aatcaatcac
     3421 tccaccaaaa cgcattcgtt tttgggattc accccgcggt tcgcaatagg tttctttttc
     3481 aaattattag cgttataaat agaaaaatgg tggagtttca ataaaaatga taatttacaa
     3541 agtgtatttt gattacatgt actcaaaaag gttgaaattt ttcataccag ttttccggaa
     3601 atccatctga tatcattatc gtatttttct ttttaaaaat gttttcaaaa aaaaacaaaa
     3661 tatagctggt tatttggcac cctctaatta ccattttcct gtcacaccac actcttttcc
     3721 ttcttccact tctttccgtt ttcagccgct tccaaccaaa ccgatatgaa agccgagcaa
     3781 caacgatcga ttccacccgg ctcggcgacc ttcccgtcgc aggtgagact cagaaaacta
     3841 gagaaaccgc tcaactaact cttgatatcc gatttcattc ttttctcttt tctttttgtt
     3901 gaacttttcc atatttccag atgccacggc caccaccaag caccgaacaa ggaatcacaa
     3961 cggaatcgga gcttgcgaag aaagctcaaa tcactccgaa cgacgtttta gcacttccgg
     4021 gaatcactca aggtatcttg ctccctttgg atttcagttt aaaacataaa tttaggattc
     4081 ttatgctccc catctgcgaa catctataac atcgagttca ccaagttcca aatccgtgat
     4141 ctggacactg agcaagtgct tttcgagatc gccaaaccgg agaacgatca ggagaatgat
     4201 gagtcgccac aggagtcggc aagatacgtg cgttatagat ttgctccaaa ctttttgaaa
     4261 ctcaaaacgg tcggagcaac tgtggaattc aaagtaggag acatcccaat ccatcatttc
     4321 cgaatgatcg aacgtcactt cttcaaagat cgccttctga agtgttttga ctttgaattc
     4381 ggattctgta ttccgaattc acgaaacaac tgtgaacata tctatgagtt ccctcaactc
     4441 tctcaacaac tcagtgagtc attattctaa aaagtacaat tcaaaagact aatctctttt
     4501 tcagtggacg acatgatcaa caatccaaac gagactcgtt ctgacagctt ctatttcgtc
     4561 gataacaaac tcgtcatgca caacaaagcc gactactcat atgatgcata aatatttaat
     4621 acaaaaatgt tctggataat tattctgtcg aatagaaaaa aaactccaaa tgtgattaaa
     4681 ttccaataat tcctgtctag tttgttcctt ccccttcccc ttctcatgtt caatgcattc
     4741 ctaagctttt cagttccccc cttgtttcta tattttttcg ctgtcctgtc acactcgcta
     4801 aaaacactaa tcacacggaa atctgttttc aataaaaact ccaactttaa ctcattttca
     4861 atttcaactg aaagattttt tcattagaga atgtctagaa ctaggcccgg gctacgtaat
     4921 acgactcact taagggtgcc ggatctgcgg cttactcacc agacctggcc ccatccgatt
     4981 accacctatt cgcttcgatg ggacacgcac tcgctgagca gcgcttcgat tcttacgaaa
     5041 gtgtgaaaaa atggctcgat gaatggttcg ccgcaaaaga cgatgagttc tactggcgtg
     5101 gaatccacaa attgcccgag agatgggaaa aatgtgtagc tagcgacggc aaatactttg
     5161 aataaatgat tttttctttt tccacaaaat ttaacgtgtt ttttgattta aaaaaaacga
     5221 catttcatac ttgtacacct gatatctaga tacctaggtg agctctggta cccccctggc
     5281 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     5341 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
     5401 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     5461 agtgcaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     5521 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5581 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     5641 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5701 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5761 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5821 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5881 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5941 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     6001 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     6061 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6121 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6181 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6241 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6301 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6361 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6421 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6481 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6541 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6601 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6661 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6721 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6781 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6841 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6901 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6961 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     7021 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7081 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7141 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7201 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7261 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7321 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7381 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7441 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggtctt ttgctggcct
     7501 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7561 tttgagtgag ctgatacc
//
