LOCUS       sequence-42169-e        7861 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 7861)
  AUTHORS   Frokjaer-Jensen C, Davis MW, Ailion M, Jorgensen EM
  TITLE     Improved Mos1-mediated transgenesis in C. elegans.
  JOURNAL   Nat Methods. 2012 Jan 30;9(2):117-8. doi: 10.1038/nmeth.1865.
  PUBMED    22290181
REFERENCE   2  (bases 1 to 7861)
  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"; 
            journalName: "Nat Methods"; date: "2012-01-30- 30"; volume: "9"; 
            issue: "2"; pages: "117-8"
COMMENT     Sequence Label: pCFJ352 - MCS(ttTi4348, I)
FEATURES             Location/Qualifiers
     source          1..7861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(21..40)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      834..1422
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1323..1342
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1559..1579
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    3083..3107
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     3114..3130
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3114..3130
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3326..3343)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3326..3342)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5283..5294
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(5440..5464)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(7070..7090)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(7294..7313)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7413..7435
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7473..7491)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7559..7663
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7664..7732,7733..7861,1..663)
                     /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"
ORIGIN
        1 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
       61 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
      121 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
      181 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
      241 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
      301 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
      361 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
      421 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
      481 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
      541 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
      601 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
      661 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
      721 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
      781 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
      841 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
      901 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
      961 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     1021 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     1081 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     1141 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     1201 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     1261 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     1321 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     1381 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     1441 tttttacggt tcctggtctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     1501 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccat cgaattatca
     1561 actatgtata ataaagttgg cgctttaatt agggtgcaag ttttttataa tttttaatcc
     1621 cattaaaatt caaagcagaa aggttctgat gctctacgca aaatatcaaa atcacaattt
     1681 atgacaatat tgttacaaaa atccaaaaaa aaaattgcag ttcgaaatgg tgattgaaga
     1741 aaacggatta cgtgaagcta ctcgacgaat catatcctat ttggaaaaat atattcatgc
     1801 actgcactat catcataagg atgatgagat ttaataattt ttgtaattta tcttacaatt
     1861 agcattttat ttaattttaa aaaaatttga atgggaacat aaagtcaaat tcctgagctg
     1921 catggctaag ggtgccctcg aaaattttcc gattgttcga tttatttcag agaaattcgg
     1981 aaaatgcgaa atcgaaaaaa agtgtagaaa atcaaaaaat gcaaaaattt ataccaaacc
     2041 gtaaaaaatt accaaaattt aaagatattt tacggaaact tacgcttgag ttattcaaac
     2101 acataactga tattataaca ttaatactat attaatacta gattaatatg tactatagca
     2161 attttcaaaa tattttcaat tagcaaaatg ttgccgatca tttttctaca tttacaatta
     2221 aaattaaaat ttgataaccc tgatccggaa aaagttacgg cagctgacat ttgtctgatt
     2281 tttgactaga atcgctaagt ggcccacaat ttttaaacta ggtgttttca cgaaacaaag
     2341 ttttctattt ttttgcaatt ttttatcctg tcacaaaagg accataaaaa ttttatacaa
     2401 aagattaaac atgaaactca aatttcaaaa attagagtgt ttcgtaaaaa gtaacgtaca
     2461 tatggtgcca aaatggccaa atatcatagt aaaatttttc aaaacatttt tgaaaaaagt
     2521 ggcaattact gagtttttgc cacttttttt tagtaaacta tagctctctc gcatactcga
     2581 attcctacaa aaatttatat ttgaaatata agtaccataa aatccggaga aaaacttcca
     2641 aaaaaatggc aaaaactaac attcttccaa ttttcaaaaa atcatataaa atttggaatt
     2701 ttttaaagaa tttttatact atgatatttg gacctttaag ctgcgttttt gtagttgaca
     2761 atttaaaaaa aaactccaaa ttgctttgaa aactaccaaa atgagtaccc tgataaccat
     2821 attgaaaaac catcaaatgg gtaatgtcta gaaaaatgaa cgaaaaatta gagaaaatag
     2881 gaaatgagat atgttcttct tcctattctt tatcttctaa gcttctcaca ccttttctct
     2941 cgtctctcca cgatttacac actatttgca atgaatctca ctctgatgag cgtatctatc
     3001 aagtccttgt ctcttctttt gacaaaactt caaagaaatc gccgacttgc gaggaggggg
     3061 gagcgaaaaa taattgatta ctgccacttt gtacaagaaa gctgggtcac gggcaggaaa
     3121 cagctatgac catgattacg ccaagctacg taatacgact cactagtggg cagatcttcg
     3181 aatgcatcgc gcgcaccgta cgtctcgagg aattcctgca ggatatctgg atccacgaag
     3241 cttcccatgg tgacgtcacc ggttctagat acctaggtga gctctggtac cctctagtca
     3301 aggccttaag tgagtcgtat tacggactgg ccgtcgtttt acacccgtgc agttgaaatt
     3361 gaaaatgagt taaagttgga gtttttattg aaaacagatt tccgtgtgat tagtgttttt
     3421 agcgagtgtg acaggacagc gaaaaaatat agaaacaagg ggggaactga aaagcttagg
     3481 aatgcattga acatgagaag gggaagggga aggaacaaac tagacaggaa ttattggaat
     3541 ttaatcacat ttggagtttt ttttctattc gacagaataa ttatccagaa catttttgta
     3601 ttaaatattt atgcatcata tgagtagtcg gctttgttgt gcatgacgag tttgttatcg
     3661 acgaaataga agctgtcaga acgagtctcg tttggattgt tgatcatgtc gtccactgaa
     3721 aaagagatta gtcttttgaa ttgtactttt tagaataatg actcactgag ttgttgagag
     3781 agttgaggga actcatagat atgttcacag ttgtttcgtg aattcggaat acagaatccg
     3841 aattcaaagt caaaacactt cagaaggcga tctttgaaga agtgacgttc gatcattcgg
     3901 aaatgatgga ttgggatgtc tcctactttg aattccacag ttgctccgac cgttttgagt
     3961 ttcaaaaagt ttggagcaaa tctataacgc acgtatcttg ccgactcctg tggcgactca
     4021 tcattctcct gatcgttctc cggtttggcg atctcgaaaa gcacttgctc agtgtccaga
     4081 tcacggattt ggaacttggt gaactcgatg ttatagatgt tcgcagatgg ggagcataag
     4141 aatcctaaat ttatgtttta aactgaaatc caaagggagc aagatacctt gagtgattcc
     4201 cggaagtgct aaaacgtcgt tcggagtgat ttgagctttc ttcgcaagct ccgattccgt
     4261 tgtgattcct tgttcggtgc ttggtggtgg ccgtggcatc tggaaatatg gaaaagttca
     4321 acaaaaagaa aagagaaaag aatgaaatcg gatatcaaga gttagttgag cggtttctct
     4381 agttttctga gtctcacctg cgacgggaag gtcgccgagc cgggtggaat cgatcgttgt
     4441 tgctcggctt tcatatcggt ttggttggaa gcggctgaaa acggaaagaa gtggaagaag
     4501 gaaaagagtg tggtgtgaca ggaaaatggt aattagaggg tgccaaataa ccagctatat
     4561 tttgtttttt tttgaaaaca tttttaaaaa gaaaaatacg ataatgatat cagatggatt
     4621 tccggaaaac tggtatgaaa aatttcaacc tttttgagta catgtaatca aaatacactt
     4681 tgtaaattat catttttatt gaaactccac catttttcta tttataacgc taataatttg
     4741 aaaaagaaac ctattgcgaa ccgcggggtg aatcccaaaa acgaatgcgt tttggtggag
     4801 tgattgattc gaatcgaaga agaaaaagaa gaagacgtgg aatagagagc tcactcttaa
     4861 ccgagcagca cacaccgaca gaaaaaaaaa tgaaatgaat gagggtcttc ttcttcttct
     4921 tcttcgaatg attgacagaa atgggaaaaa gaggaagatt gagaagggaa aaaggaagga
     4981 gaaaagaagc agaagaagac gtcagagagg agaggaacga gcggaaaagc agcgggcgca
     5041 agtcatagaa gtagcagagc tggggagaag aagacactat ccaagaaagg aatgacgaga
     5101 gagtatgcaa aggggtatag ggtgcagaca gaataggaac agaataacag atgatgagcc
     5161 aagaagagtt gaaaagggcg atgaatttgt catgtaactt aatttgggtc aatttgagca
     5221 tgatgaattg aaatcatccc ttgttgggag ttaataaccg gtttgttatc agaaaccctg
     5281 taatagaagg gcgccctaac tttgagccaa ttcatcccgg tttctgtcaa atatatcaaa
     5341 aagtggtcaa ctgacaaatt gtttttgata ttataataaa cattttatcc gttaacaatt
     5401 ttcgaatact ttttacaagg acttggataa attggctcaa gcctgctttt ttgtacaaac
     5461 ttgcatcttt attttttaat tcgattttct gatggcaatg actaaccgat tttcgggata
     5521 attgagatga ggggaggtat tttgggaatt gacagggagg tatttctgca gattttgtta
     5581 tcagagtgta ctgtagctct aaaaccaata cttgacataa tacttgaatc tatacagttg
     5641 ataagttttt ttgcagtttc aagttgaata aagttatagt ggccgacatc ttagaggtct
     5701 aaaaatgaga ttttggtcaa atttgagcta gtccaattat acaactaatc gcagttcttg
     5761 cgcccgacac tttacgggat ttcaattgta tatgttttgt ttcaaggttt catctgttca
     5821 gacaatgcga agcattttgt tggcaggttt acaaagtttt agcggccgac acctcgcggg
     5881 tttctaagat aagatttttg cgaagtcttc agccatctgt aagctacact agcggtcgac
     5941 atctcgcggg ttaaatctag cttttgaaaa actatccaaa cactagaaat atttaaattt
     6001 taattttaat ttttgaaaaa aaattccatt ttaaattccg gttttcaaaa aaaaacaata
     6061 gatcaaaaaa ttttaacgta tttttttgca attttttcaa aatgcataac tttcagaata
     6121 tttctcacct cttcgcatgg tgaaaaccgt caagagtttt tttttcaaaa taaatggaaa
     6181 aatcgaaaat tttagtttat caatttttcg tatattcgag aaaaatctta aaattctggt
     6241 catttttctt cttcttagaa aaatattgta tgttttcttt ctcattacca taaacaaaat
     6301 ttgataaaat attgaatttt gctgctattt ttcttaaaat atttttgtcg attttcggtt
     6361 ttcataaatg tctaaaattc gttgcgtaac tttattgaat tatcagcaaa acaaacaaat
     6421 aaaaaacgaa aaatatccat tttaaaaaat tagataaatg aatcaacacg tccctgaatg
     6481 tagggacgaa tgtgcggaag agactcaaaa gtctggcaaa aagttcggag cctctcgaaa
     6541 tgagccagat agaagctatc gtagcactgc tgacaccgag ttagaaactc gacgacagcg
     6601 atgtcagccc atgaatatcg atctccaatc atgaactttt gctcgccgag caactcttca
     6661 tattctctga gcaatgggaa gagtgactcc tctttgaata cattccagca ttctttctga
     6721 aaaggaagta ttgaatttta tgaggctatc tacctgcctg actaccgcct atttccatgc
     6781 gcagtaggca catgtgtaga cataggcagg cattatcgcg cctcccttca aactacctac
     6841 taggtactag gtagttcgaa aggagacgcg ataatgcctg cctataccta cacacgttcc
     6901 tactgcacat ggaaataggc ggcaggcagg caggtggata gcttcaaggt agtgcctacc
     6961 ctcgcgtcat agttcttcgt cagtgttgct ctcaaaattg ggttcatcgc tgtgttgagc
     7021 tcctcgagtt tttgaatcac tttatcaact tcttcctgat ccccacttgc aacttttcta
     7081 tacaaagttg atagcttggc ggatccctgg cgttacccaa cttaatcgcc ttgcagcaca
     7141 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     7201 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg
     7261 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     7321 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     7381 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     7441 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     7501 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     7561 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     7621 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     7681 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     7741 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     7801 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     7861 g
//
