LOCUS       sequence-42170-e        6960 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6960)
  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 6960)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pCFJ353 - MCS(ttTi4391, I)
FEATURES             Location/Qualifiers
     source          1..6960
                     /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    3060..3084
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     3091..3107
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3091..3107
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3303..3320)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3303..3319)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5260..5271
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(5417..5441)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6169..6189)
                     /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(6393..6412)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6512..6534
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6572..6590)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6658..6762
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6763..6831,6832..6960,1..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 ataaagttgt ccatcagact ccaatgcaag tgcgaatgta aaacctgcga
     1621 atgtagcact tagtaagaat ggtcaatggt tctacttgac agctcgacag tatgaaaatg
     1681 gaatctggta tcatatatcc ggagcaagtg ccatatctaa aagacatgtt cttactactt
     1741 cccgagttgt catgtcaagt aagagaaaat ggagggctaa taatcaatcg gtagactgta
     1801 aaggtgactt gaagtaaggc taaatttgtt atataatatt gatttgaatc tgtgttcaga
     1861 tacgtcgagg ttccagcagg atttctggaa aaaaacatgt ttagacgttt tcaatgtgta
     1921 atgaacggga ccgactgcca tgatatagct actcctgtga aagcttatat tttgaactac
     1981 tgcccgaaca tgacagagca tgtggcctat catccaatga ttatcgagtt tgatgacact
     2041 ttcacgaaca gtacaccttg tctatcagat gaagttaata ttggtaatgg ggtttatgtg
     2101 catggttatg aagacaatgg taatgggaaa aataacggcg ttggattgaa gcatcgactt
     2161 ctttgggtaa tcttgaatct aatagatttc aaagagcaag tacatatttg caggtaatag
     2221 acttacaaga gaaactagtc attactgaaa gctttcatgg ggaatatgag tttggaagcc
     2281 ctttatccag agtgattggc agaaaatgga caattgttgg gctgggtaca acttgtaagt
     2341 tttagattta aggttgaatt gtccgatctt ttttcagata tggatttcac ttctaatgtg
     2401 ttcttccgca ggtctgaagg cttgcagcgt cgtagaatta gaaactcttc aaattagaga
     2461 tcctatgagg tcgagcgccg cgccggcctc acggcgaaac cccggacgtg tcgcttccaa
     2521 ataactacct tttcgcaaaa aaaggagcaa aaaatcacaa gaaaggagcc ggggtgagct
     2581 ttttctatga agaaaaacaa tttaaaaaaa aattgaattt cagggcaaga agaaggaggc
     2641 agctgctgct ggacatgcct aacatttctg ataattttca aatttttcgc aataaatatt
     2701 tttggtttgt aaaattagtt tcttggaaac aagcagggta ctgtaggagt tatgtagaag
     2761 aactgtaggg gtactgtggg ggcattgtac tcagtaaacc cgttcagaat atacccagat
     2821 tcaaacaaaa ttagaaccgt acaagcctat attgtgaatt tatctgcaca aatttaattt
     2881 ggatgtacac aaaatcaatt tggatctaca caaacctaca caaattcagt ttacatctac
     2941 tcaaacttgc aatctatgca aattaagttt ggatctagac aatcttggtt tacatctgcc
     3001 caaaattaga atttttcata gtataggctt gtacggtttt aaatgggttt gaatctgggt
     3061 ccactttgta caagaaagct gggtcacggg caggaaacag ctatgaccat gattacgcca
     3121 agctacgtaa tacgactcac tagtgggcag atcttcgaat gcatcgcgcg caccgtacgt
     3181 ctcgaggaat tcctgcagga tatctggatc cacgaagctt cccatggtga cgtcaccggt
     3241 tctagatacc taggtgagct ctggtaccct ctagtcaagg ccttaagtga gtcgtattac
     3301 ggactggccg tcgttttaca cccgtgcagt tgaaattgaa aatgagttaa agttggagtt
     3361 tttattgaaa acagatttcc gtgtgattag tgtttttagc gagtgtgaca ggacagcgaa
     3421 aaaatataga aacaaggggg gaactgaaaa gcttaggaat gcattgaaca tgagaagggg
     3481 aaggggaagg aacaaactag acaggaatta ttggaattta atcacatttg gagttttttt
     3541 tctattcgac agaataatta tccagaacat ttttgtatta aatatttatg catcatatga
     3601 gtagtcggct ttgttgtgca tgacgagttt gttatcgacg aaatagaagc tgtcagaacg
     3661 agtctcgttt ggattgttga tcatgtcgtc cactgaaaaa gagattagtc ttttgaattg
     3721 tactttttag aataatgact cactgagttg ttgagagagt tgagggaact catagatatg
     3781 ttcacagttg tttcgtgaat tcggaataca gaatccgaat tcaaagtcaa aacacttcag
     3841 aaggcgatct ttgaagaagt gacgttcgat cattcggaaa tgatggattg ggatgtctcc
     3901 tactttgaat tccacagttg ctccgaccgt tttgagtttc aaaaagtttg gagcaaatct
     3961 ataacgcacg tatcttgccg actcctgtgg cgactcatca ttctcctgat cgttctccgg
     4021 tttggcgatc tcgaaaagca cttgctcagt gtccagatca cggatttgga acttggtgaa
     4081 ctcgatgtta tagatgttcg cagatgggga gcataagaat cctaaattta tgttttaaac
     4141 tgaaatccaa agggagcaag ataccttgag tgattcccgg aagtgctaaa acgtcgttcg
     4201 gagtgatttg agctttcttc gcaagctccg attccgttgt gattccttgt tcggtgcttg
     4261 gtggtggccg tggcatctgg aaatatggaa aagttcaaca aaaagaaaag agaaaagaat
     4321 gaaatcggat atcaagagtt agttgagcgg tttctctagt tttctgagtc tcacctgcga
     4381 cgggaaggtc gccgagccgg gtggaatcga tcgttgttgc tcggctttca tatcggtttg
     4441 gttggaagcg gctgaaaacg gaaagaagtg gaagaaggaa aagagtgtgg tgtgacagga
     4501 aaatggtaat tagagggtgc caaataacca gctatatttt gttttttttt gaaaacattt
     4561 ttaaaaagaa aaatacgata atgatatcag atggatttcc ggaaaactgg tatgaaaaat
     4621 ttcaaccttt ttgagtacat gtaatcaaaa tacactttgt aaattatcat ttttattgaa
     4681 actccaccat ttttctattt ataacgctaa taatttgaaa aagaaaccta ttgcgaaccg
     4741 cggggtgaat cccaaaaacg aatgcgtttt ggtggagtga ttgattcgaa tcgaagaaga
     4801 aaaagaagaa gacgtggaat agagagctca ctcttaaccg agcagcacac accgacagaa
     4861 aaaaaaatga aatgaatgag ggtcttcttc ttcttcttct tcgaatgatt gacagaaatg
     4921 ggaaaaagag gaagattgag aagggaaaaa ggaaggagaa aagaagcaga agaagacgtc
     4981 agagaggaga ggaacgagcg gaaaagcagc gggcgcaagt catagaagta gcagagctgg
     5041 ggagaagaag acactatcca agaaaggaat gacgagagag tatgcaaagg ggtatagggt
     5101 gcagacagaa taggaacaga ataacagatg atgagccaag aagagttgaa aagggcgatg
     5161 aatttgtcat gtaacttaat ttgggtcaat ttgagcatga tgaattgaaa tcatcccttg
     5221 ttgggagtta ataaccggtt tgttatcaga aaccctgtaa tagaagggcg ccctaacttt
     5281 gagccaattc atcccggttt ctgtcaaata tatcaaaaag tggtcaactg acaaattgtt
     5341 tttgatatta taataaacat tttatccgtt aacaattttc gaatactttt tacaaggact
     5401 tggataaatt ggctcaagcc tgcttttttg tacaaacttg tatgaacggt tttactgagt
     5461 aggggtgttg catctatacc gtagagaaac cgaaggggca ctgtagcggt actgcaagat
     5521 tattgtttag gggtactgta aggataccat gggagtactg tagaagtact gtaggggtgc
     5581 tgtagggata ctgtagggat actgttgtag tgctgtaggg gtactgtagc gacactgtaa
     5641 ctataacgca gaggaaccgt agaggtactg tagatgttac cgtagaggta ccggtactgt
     5701 aagggtactg tcggggtaca gaaatagtaa tgtagaggta acgtatggat attgtgttgg
     5761 ggtactgtag cggcactgta ggggtgttgc atctataccg tagagaaacc gaaggggcac
     5821 tgtagcggta ctgcaagatt attgtttagg ggtactgtaa ggataccatg ggagtagtgt
     5881 agaagtactg taggggtact gtagggggac tgtaaggata ctgtagaggt actgtagagg
     5941 tacctaagtg gtattgtgag gtactgtagg ggtactgtaa gattattgta gaggaactgt
     6001 agagatactg tcgaggtact gtggggttac tgtaggagtt ataacaagct ttattctcaa
     6061 taataatagt caaaatccgc atacatcgga ggaatcatca atagctgctt gatacactcc
     6121 tcctcgattt cactgcgatg tctgtaatct tgttgaaaac ggtgatcaca acttttctat
     6181 acaaagttga tagcttggcg gatccctggc gttacccaac ttaatcgcct tgcagcacat
     6241 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     6301 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
     6361 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     6421 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     6481 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     6541 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     6601 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     6661 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6721 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6781 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6841 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6901 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
//
