LOCUS       sequence-60719-e        6514 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 6514)
  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 6514)
  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: pCFJ1201
FEATURES             Location/Qualifiers
     source          1..6514
                     /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"
     misc_feature    341..448
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     357..373
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     358..374
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(408..424)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(408..424)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    complement(448..468)
                     /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(476..493)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(476..492)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(498..509)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      3932..4255
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(4399..4418)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4518..4540
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4578..4596)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4664..4768
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4769..4837,4838..5629)
                     /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(4987..5006)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5800..6388
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6289..6308
                     /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 ggtaccgggc cccccctcga
      361 ggtcgacggt atcgataagc ttgatatcga attcctgcag cccgggggat ccactagttc
      421 tagagcggcc gccaccgcgg tggagctcaa ctttattata catagttgat aattcactgg
      481 ccgtcgtttt acatggcacg cccttctatt acagggtttc tgataacaaa ccggttatta
      541 actcccaaca agggatgatt tcaattcatc atgctcaaat tgacccaaat taagttacat
      601 gacaaattca tcgccctttt caactcttct tggctcatca tctgttattc tgttcctatt
      661 ctgtctgcac cctatacccc tttgcatact ctctcgtcat tcctttcttg gatagtgtct
      721 tcttctcccc agctctgcta cttctatgac ttgcgcccgc tgcttttccg ctcgttcctc
      781 tcctctctga cgtcttcttc tgcttctttt ctccttcctt tttcccttct caatcttcct
      841 ctttttccca tttctgtcaa tcattcgaag aagaagaaga agaagaccct cattcatttc
      901 attttttttt ctgtcggtgt gtgctgctcg gttaagagtg agctctctat tccacgtctt
      961 cttctttttc ttcttcgatt cgaatcaatc actccaccaa aacgcattcg tttttgggat
     1021 tcaccccgcg gttcgcaata ggtttctttt tcaaattatt agcgttataa atagaaaaat
     1081 ggtggagttt caataaaaat gataatttac aaagtgtatt ttgattacat gtactcaaaa
     1141 aggttgaaat ttttcatacc agttttccgg aaatccatct gatatcatta tcgtattttt
     1201 ctttttaaaa atgttttcaa aaaaaaacaa aatatagctg gttatttggc accctctaat
     1261 taccattttc ctgtcacacc acactctttt ccttcttcca cttctttccg ttttcagccg
     1321 cttccaacca aaccgatatg aaagccgagc aacaacgatc gattccaccc ggctcggcga
     1381 ccttcccgtc gcaggtgaga ctcagaaaac tagagaaacc gctcaactaa ctcttgatat
     1441 ccgatttcat tcttttctct tttctttttg ttgaactttt ccatatttcc agatgccacg
     1501 gccaccacca agcaccgaac aaggaatcac aacggaatcg gagcttgcga agaaagctca
     1561 aatcactccg aacgacgttt tagcacttcc gggaatcact caaggtatct tgctcccttt
     1621 ggatttcagt ttaaaacata aatttaggat tcttatgctc cccatctgcg aacatctata
     1681 acatcgagtt caccaagttc caaatccgtg atctggacac tgagcaagtg cttttcgaga
     1741 tcgccaaacc ggagaacgat caggagaatg atgagtcgcc acaggagtcg gcaagatacg
     1801 tgcgttatag atttgctcca aactttttga aactcaaaac ggtcggagca actgtggaat
     1861 tcaaagtagg agacatccca atccatcatt tccgaatgat cgaacgtcac ttcttcaaag
     1921 atcgccttct gaagtgtttt gactttgaat tcggattctg tattccgaat tcacgaaaca
     1981 actgtgaaca tatctatgag ttccctcaac tctctcaaca actcagtgag tcattattct
     2041 aaaaagtaca attcaaaaga ctaatctctt tttcagtgga cgacatgatc aacaatccaa
     2101 acgagactcg ttctgacagc ttctatttcg tcgataacaa actcgtcatg cacaacaaag
     2161 ccgactactc atatgatgca taaatattta atacaaaaat gttctggata attattctgt
     2221 cgaatagaaa aaaaactcca aatgtgatta aattccaata attcctgtct agtttgttcc
     2281 ttccccttcc ccttctcatg ttcaatgcat tcctaagctt ttcagttccc cccttgtttc
     2341 tatatttttt cgctgtcctg tcacactcgc taaaaacact aatcacacgg aaatctgttt
     2401 tcaataaaaa ctccaacttt aactcatttt caatttcaac tgaaagattt tttcattaga
     2461 gaatgtctag aactaggccc gggctacgta atacgactca cttaagggtg ccggatctgc
     2521 ggcttactca ccagacctgg ccccatccga ttaccaccta ttcgcttcga tgggacacgc
     2581 actcgctgag cagcgcttcg attcttacga aagtgtgaaa aaatggctcg atgaatggtt
     2641 cgccgcaaaa gacgatgagt tctactggcg tggaatccac aaattgcccg agagatggga
     2701 aaaatgtgta gctagcgacg gcaaatactt tgaataaatg attttttctt tttccacaaa
     2761 atttaacgtg ttttttgatt taaaaaaaac gacatttcat acttgtacac ctgatatcta
     2821 gatacctagg tgagctctgg tacccccctg gcgttaccca acttaatcgc cttgcagcag
     2881 agtcgtatta cgtagcccgg cctgcaggtc gactctagag gatcaccaaa aacggaacgt
     2941 tgagctggac ggaaatagtg gtaaagtgac atgattatag tttgaagatt tctaatttca
     3001 caattagagc aaatgttgtt cggtatttat tttcaacggt atttatacta ttttccacct
     3061 ttttctagaa cattcgagct gcttgttgca aaaggagggc gactcacatt cggtacatgg
     3121 aaaagtagtg tacacaataa agagacccag atacattttc cgtctgcgtc tctttgcacc
     3181 caccgggagt attttcaaac gaatgcatct aggaccttct agaacattct gtaaggctgc
     3241 agaatgcggg tatataagga aagcgggctc agaggaagcc aacacgcttt gttctagtgc
     3301 atctaaaaaa cttcgaaaat cctcatcgga tcctatcgat tcgcggccgc tgtacacccg
     3361 ggtgccctat agtgagtcgt attggtaccg gtagaaaaaa tatgcgcttt gatttccaaa
     3421 acttaaaatt ttcgatgatt ttcatatttt tgtggaacat catagtcgga tttttacttg
     3481 ctcttgtaaa gattttcaaa atctacatgt atcttgatct gcctgaaaat gtatgggtaa
     3541 atcggtttgc gcatgttatt gctcttattg gatcaattgc ttcattgtgg ttaatttttt
     3601 ttagcccatt tgaaattaag caaggaagat tttctgtctc gtcaaaaact atcctgcttc
     3661 ttgtttgcaa gggtttgatt ggcttgatat ttcttgcaca gattttcgcc catgtccttt
     3721 acattgaccc atgccagaat atgctcttat cattgaactc tgcactgttc tccttgctgc
     3781 tgaattatta tgagtctcgt ggtgcgatcc atccgttgat ggaattagtt caaacagctg
     3841 gcctgagacc aactttgtgc gctgctgtac gatttaattg tctcaactca actgctcata
     3901 ttgatccaag tgttgaaaat ccatgagata aatgcaaaat cctttcaagc attcccttct
     3961 tctctatcac tcttctttct ttttgtcaaa aaattctctc gctaatttat ttgctttttt
     4021 aatgttatta ttttatgact ttttatagtc actgaaaagt ttgcatctga gtgaagtgaa
     4081 tgctatcaaa atgtgattct gtctgatgta ctttcacaat ctctcttcaa ttccattttg
     4141 aagtgcttta aacccgaaag gttgagaaaa atgcgagcgc tcaaatattt gtattgtgtt
     4201 cgttgagtga cccaacaaaa agaggaaact ttattgtgcc gccaagaaaa aagtctcaag
     4261 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     4321 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4381 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     4441 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     4501 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     4561 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4621 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4681 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4741 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4801 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4861 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4921 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4981 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5041 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5101 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5161 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5221 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5281 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5341 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5401 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5461 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5521 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5581 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5641 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5701 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5761 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5821 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5881 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5941 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6001 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6061 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6121 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6181 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6241 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6301 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6361 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6421 ggtcttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6481 taaccgtatt accgcctttg agtgagctga tacc
//
