LOCUS       sequence-42172-e        7606 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 7606)
  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 7606)
  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: pCFJ356 - MCS(cxTi10816, IV)
FEATURES             Location/Qualifiers
     source          1..7606
                     /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    2999..3023
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     3030..3046
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3030..3046
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3242..3259)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3242..3258)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5199..5210
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(5356..5380)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6815..6835)
                     /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(7039..7058)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7158..7180
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7218..7236)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7304..7408
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7409..7477,7478..7606,1..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 ataaagttgc ggtacccaag gcgacaacgt aagtgactga aggcattaaa
     1621 ggtacatgcg tccgcgcaag tgtccgcttt tgctggccta acaagtaaga gaaacaagta
     1681 agaaagagta agagaaaact cacccgaggt gtctcttcta gcaaccgcct attcagataa
     1741 ttttaagcca gttcgagagt tatttgtcgc atggtataga agctcgttat ttacattaaa
     1801 tttttagtct tcccgcagaa gtggacgatg cgatgggcta cttcaaaaat ctgacgccac
     1861 gtgtcaagga ccttcacaag agcatgacaa acttggaaga tattagcaag tggcagaaga
     1921 aggccagttt ctctggcacc cttgagaatt actcgcgtct cggtgacaag atcaatgtga
     1981 aaccatttat ggatgctgtt gatgctagaa tgggtgccga agctgatgcc gtgaaagggg
     2041 tataaagcta ctgttgcacg acgactcaca gactaatcat tccaggtcct cgcgatttgt
     2101 gaaaaataca agtcattcta ccaaaacgag ggaaaacttc acgcggacag tatcgccaat
     2161 ttgaatagga ctcggctcga catggacagt gcggcggata aatatgcgaa caacgagact
     2221 gaagttaaca agactcgttt ggataacagt accacggaat ttgaagtggc ttgtgagagg
     2281 tttgtaggtc tttcgtaggc acgatatcct tatttgaatg tgtttcagaa tgcgagaact
     2341 ggcgaacgga atcaagacaa ttgaatcgaa ccattcttcc tggcaagacg gtcttatgaa
     2401 ggagattaaa gtggcgttgc gtaaataaag aaaggatcct ctttgaaaat aaagattttc
     2461 tacattcact tattttttta gctttatact tttcagatcg aatgaactgt ttccaacgtt
     2521 tccatattaa ttttttttag ttaaaaaagt tgggtattaa cagttttgtt aacagttttg
     2581 tcttccgaca gatgtgagac ggaaaaatgg ccaaaattcg agattttgac taaaatcagt
     2641 gtagcttttt caaaattttg aaccggcatg aaaaagtttg aaaagttttc gagaagtttc
     2701 aatccgttca tttgagccca tgttgggtct ataagatcaa aatcgcccga actgttagtc
     2761 ctcctttaag ttacgattat taacaaaatg tcgcctaagt ttagtttcac acttgtctca
     2821 cactagtcga tgaactgtgt gactactgta gtgagtacag tagggcccgt gtaaaaatac
     2881 aaaaaccaag aaaaagtaaa ccaaaataaa aaacgaattt attgttcact acaaccaaac
     2941 tgcaaagttc ttttcttacc catcagctca atcgtgtact tgcgtggaaa ccctaggagc
     3001 cactttgtac aagaaagctg ggtcacgggc aggaaacagc tatgaccatg attacgccaa
     3061 gctacgtaat acgactcact agtgggcaga tcttcgaatg catcgcgcgc accgtacgtc
     3121 tcgaggaatt cctgcaggat atctggatcc acgaagcttc ccatggtgac gtcaccggtt
     3181 ctagatacct aggtgagctc tggtaccctc tagtcaaggc cttaagtgag tcgtattacg
     3241 gactggccgt cgttttacac ccgtgcagtt gaaattgaaa atgagttaaa gttggagttt
     3301 ttattgaaaa cagatttccg tgtgattagt gtttttagcg agtgtgacag gacagcgaaa
     3361 aaatatagaa acaagggggg aactgaaaag cttaggaatg cattgaacat gagaagggga
     3421 aggggaagga acaaactaga caggaattat tggaatttaa tcacatttgg agtttttttt
     3481 ctattcgaca gaataattat ccagaacatt tttgtattaa atatttatgc atcatatgag
     3541 tagtcggctt tgttgtgcat gacgagtttg ttatcgacga aatagaagct gtcagaacga
     3601 gtctcgtttg gattgttgat catgtcgtcc actgaaaaag agattagtct tttgaattgt
     3661 actttttaga ataatgactc actgagttgt tgagagagtt gagggaactc atagatatgt
     3721 tcacagttgt ttcgtgaatt cggaatacag aatccgaatt caaagtcaaa acacttcaga
     3781 aggcgatctt tgaagaagtg acgttcgatc attcggaaat gatggattgg gatgtctcct
     3841 actttgaatt ccacagttgc tccgaccgtt ttgagtttca aaaagtttgg agcaaatcta
     3901 taacgcacgt atcttgccga ctcctgtggc gactcatcat tctcctgatc gttctccggt
     3961 ttggcgatct cgaaaagcac ttgctcagtg tccagatcac ggatttggaa cttggtgaac
     4021 tcgatgttat agatgttcgc agatggggag cataagaatc ctaaatttat gttttaaact
     4081 gaaatccaaa gggagcaaga taccttgagt gattcccgga agtgctaaaa cgtcgttcgg
     4141 agtgatttga gctttcttcg caagctccga ttccgttgtg attccttgtt cggtgcttgg
     4201 tggtggccgt ggcatctgga aatatggaaa agttcaacaa aaagaaaaga gaaaagaatg
     4261 aaatcggata tcaagagtta gttgagcggt ttctctagtt ttctgagtct cacctgcgac
     4321 gggaaggtcg ccgagccggg tggaatcgat cgttgttgct cggctttcat atcggtttgg
     4381 ttggaagcgg ctgaaaacgg aaagaagtgg aagaaggaaa agagtgtggt gtgacaggaa
     4441 aatggtaatt agagggtgcc aaataaccag ctatattttg tttttttttg aaaacatttt
     4501 taaaaagaaa aatacgataa tgatatcaga tggatttccg gaaaactggt atgaaaaatt
     4561 tcaacctttt tgagtacatg taatcaaaat acactttgta aattatcatt tttattgaaa
     4621 ctccaccatt tttctattta taacgctaat aatttgaaaa agaaacctat tgcgaaccgc
     4681 ggggtgaatc ccaaaaacga atgcgttttg gtggagtgat tgattcgaat cgaagaagaa
     4741 aaagaagaag acgtggaata gagagctcac tcttaaccga gcagcacaca ccgacagaaa
     4801 aaaaaatgaa atgaatgagg gtcttcttct tcttcttctt cgaatgattg acagaaatgg
     4861 gaaaaagagg aagattgaga agggaaaaag gaaggagaaa agaagcagaa gaagacgtca
     4921 gagaggagag gaacgagcgg aaaagcagcg ggcgcaagtc atagaagtag cagagctggg
     4981 gagaagaaga cactatccaa gaaaggaatg acgagagagt atgcaaaggg gtatagggtg
     5041 cagacagaat aggaacagaa taacagatga tgagccaaga agagttgaaa agggcgatga
     5101 atttgtcatg taacttaatt tgggtcaatt tgagcatgat gaattgaaat catcccttgt
     5161 tgggagttaa taaccggttt gttatcagaa accctgtaat agaagggcgc cctaactttg
     5221 agccaattca tcccggtttc tgtcaaatat atcaaaaagt ggtcaactga caaattgttt
     5281 ttgatattat aataaacatt ttatccgtta acaattttcg aatacttttt acaaggactt
     5341 ggataaattg gctcaagcct gcttttttgt acaaacttgc actagtgaca cacggccaaa
     5401 tttcttattt tcctgctatt ttctccaaat atgattaaaa tttgttcatt tcaaaattgc
     5461 ctccaacttg ttttggggca aaaatttcac aaaacttcac agcaaaatgt gaaacaatat
     5521 tgctatctat tgaaaacatt aatgatagtc gagtgacttc ttgaaccaat actcatcgca
     5581 tcaaatgcct tttctcgaat actctgtaat aattggcata taaaaacata catactcatc
     5641 agtttgaaag ctaaccttta ccataatcat tactaaaatt ggatttaaag ccggacacca
     5701 agaaaaaaga tacggtagat aaactgacag atgagaagtc tcgatacgaa ataatggcat
     5761 tagaaaagac acatagaatg gaatgtggat agtgagaaat ggtagaatga actgaaaaat
     5821 tgagaattta aggagaaatg ttatttttaa gctgctttgc ttgaaacttt gaatttaaaa
     5881 ttttagaaca attactgttt caaaatattt ttatttttat ttactccatt tgaaaaaaaa
     5941 gcaacattgc ttgttcaatt tctgttattg ccttacctga accataagta ccatattcaa
     6001 ttgttgttga gacttcatcg aatttgtact ccgacttccc ttatgaattt cgtatctgat
     6061 aattgtccaa caaaccatcg caattactgt agttatcact agaacagcca ttatataaat
     6121 cgtacatgcc catgaaattg ggtcagcaac aaagtcattc aacgtgtaat catatctttc
     6181 tagtgatgca attaaatatg cagattttcc gattccattc agattgtata cgatatattc
     6241 tgatagttga tcgtcagcag catctgggac atagccaaag gtgattagaa ctgtggatat
     6301 tagagagaca ataaatgata tgattgtggt tgtaaggacc acgttttttt gtgttctcgt
     6361 tgttctgaaa gtataatttg tagattctta aactcagaat ttatcatatt tgaactccgc
     6421 tgtttttgaa ttaagtaata gctgaaacag tgaacattag tttaaaaata taattttaca
     6481 ggcacgcaca aacttcggtg tgggtacgta ttacctgcct cctattgaat attcattgtt
     6541 taaacgtttt cagaaaaaaa gtgtacatgt tttttggagt atgatgatga tgtagtcgca
     6601 aggcagatat gaaatgtagg taacacagtg cacagatgcg ttttttgcca acatcaaggg
     6661 ttgtaagtaa cttttcagag aaaaacatat agaaattttg tttcaagtat cgtatcagtt
     6721 tttttctgat taacttataa aaagaactct tcgacaattt gaatgcttaa tttaaaaaaa
     6781 ttaagtcgaa tatttcagca tgccaacccg ggagcaactt ttctatacaa agttgatagc
     6841 ttggcggatc cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     6901 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     6961 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     7021 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     7081 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     7141 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     7201 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     7261 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     7321 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     7381 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     7441 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     7501 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     7561 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatg
//
